Volume 96  ·  Issue 14  ·  2016

After the rapid development of recent decades, the training and accumulation of laboratory medicine personnel has been greatly improved. The training of talents in the whole industry not only pays attention to the study of laboratory knowledge system and the accumulation of laboratory medicine experience, but also pays special attention to the combination of laboratory and clinical practice, which greatly improves the knowledge service ability of laboratory medicine talents, and grows a group of professional laboratory talents with rich laboratory medicine experience and good communication ability with clinical disciplines. Laboratory medicine has not only cultivated undergraduate students with solid professional knowledge, but also cultivated a large number of master's and doctorate students. Since the standardized training of resident physicians in China in 2005, the standardized training of laboratory physicians has been incorporated into the national unified training system. At present, the structure of the elderly, middle-aged and young people in the laboratory medicine discipline is far better than in the past. It is no longer the state of low academic qualifications and few talents that people usually think, but a clinical professional discipline that integrates the advantages of medicine, teaching and research. It can not only complete the work before and during the analysis, but also undertake the work after the analysis, so that the laboratory medicine knowledge service system can be continuously improved and expanded.

The incidence of sexually transmitted diseases is increasing, and the prevention and treatment of sexually transmitted diseases is a problem that cannot be ignored in the world. In 2006, the State Council promulgated the Regulations on the Prevention and Control of AIDS^[1]In 2012, the Ministry of Health revised and promulgated the Administrative Measures for the Prevention and Control of STDs^[2]European and American countries have also formulated guidelines for STD management, diagnosis and treatment, etc.^[3,4]。 Laboratory diagnosis is the gold standard of STD diagnosis, and laboratory report is the decisive basis of clinical diagnosis. It is an inevitable trend to improve the quality of laboratory medicine to formulate a scientific and complete laboratory diagnosis report model. In order to ensure the accuracy and correctness of STD diagnosis and give full play to the role of laboratory medicine in the diagnosis and treatment of STD, the Expert Committee of STD Laboratory Medicine of Laboratory Physicians Branch of Chinese Medical Doctors Association referred to the Regulations on AIDS Prevention and Control promulgated by the State Council^[1]Administrative Measures for the Prevention and Treatment of STDs issued by the Ministry of Health^[2]National Clinical Laboratory Operating Procedures^[5]Infectious Diseases^[6]"Medical Microbiology"^[7]And related literature at home and abroad, and combined with the latest progress of STD laboratory medicine at home and abroad and the current situation of clinical laboratory and diagnosis at home, the expert consensus on STD testing and diagnosis report model was formulated.

Alzheimer's disease (AD) is a common progressive neurodegenerative disease, which is characterized by memory impairment, aphasia, apraxia, agnosia, impaired visual and spatial skills, executive dysfunction, and personality and behavior changes. The early symptoms lack characteristics^[1,2]。 There is no uniform standard for the detection and diagnosis of AD at home and abroad. Alzheimer's Disease Laboratory Medicine Expert Committee of Laboratory Physicians Branch of Chinese Medical Doctors Association according to the latest AD diagnostic standards at home and abroad, some authoritative research reports, etc.^[3,4,5,6,7]The AD test (auxiliary) diagnostic path was developed. Based on this path, this expert committee reached an expert consensus on the diagnostic reporting model of AD testing.

The laboratory diagnosis report is a report issued by the laboratory physician that has guiding value for clinical diagnosis, differential diagnosis and treatment, and can improve the clinical service ability and clinical diagnosis and treatment ability of the laboratory. The examination and diagnosis reports are relatively perfect in foreign medical institutions, but they are uneven in China. This consensus aims to standardize the examination report mode and lay the foundation for mutual recognition of examination reports among hospitals. Common intestinal infectious diseases include acute enteritis, viral diarrhea, Clostridium difficile infection and cholera caused by bacteria and their toxins, viruses, fungi and parasites. Clostridium difficile has become one of the main pathogens of nosocomial intestinal infections^[1,2,3,4]Clostridium difficile infection (CDI) clinically manifests as diarrhea, pseudomembranous colitis, toxic megacolon, and even death^[5,6,7,8,9]。 The main risk factors for CDI include recent use of antimicrobial drugs, and other risk factors include old age, hospitalization, immunosuppression, gastrointestinal and emergency surgery, etc^[6]。 In recent years, the incidence, recurrence and mortality of CDI have been increasing significantly^{[2,10,11,12,13]}So timely and accurate laboratory diagnosis is very necessary. Taking Clostridium difficile intestinal infection as an example, this paper aims to standardize the examination and diagnosis report mode of Clostridium difficile intestinal infection, and provide laboratory basis for clinical effective diagnosis and treatment of Clostridium difficile intestinal infection.

Clinical genetic testing has covered tumors, genetic diseases, blood diseases, infectious diseases, neuropsychiatric diseases, organ transplantation, birth defects, individualized drug therapy and other fields^{[1,2,3,4,5,6,7,8]}Clinical genetic test diagnosis reports have important reference value for body susceptibility assessment, disease diagnosis, prognosis, treatment monitoring, genetic counseling, health management and family birth plan formulation. Therefore, the clinical genetic test diagnostic report should be clear, concise, accurate and reliable, and have sufficient explanation, credibility and authority. The Molecular Diagnosis Expert Committee of Laboratory Physicians Branch of Chinese Medical Doctors Association organized relevant experts to discuss the clinical genetic test diagnosis report model and reached the following consensus.

Tumor molecule targeted therapy is a therapeutic mode in which specific genes or expression products of genes expressed by tumor cells but less expressed or not expressed by normal cells are used as therapeutic targets to kill tumor cells to the greatest extent and harm normal cells less. Human epidermal growth factor receptor 2 (HER2) is an important prognostic indicator in breast cancer patients and a major predictor of anti-HER2 drug therapy. As the first humanized monoclonal antibody against HER2, the clinical application of trastuzumab has significantly improved the prognosis of HER2-positive breast cancer patients, changed the diagnosis and treatment mode of breast cancer, and is an important breakthrough in targeted therapy of breast cancer. In recent years, new anti-HER2 therapeutics including lapatinib, pertuzumab, and Kadcyla (T-DM1) have been emerging, further improving the prognosis of HER2-positive breast cancer patients. In order to better promote the standardized HER2 standard detection, accurately assess patient prognosis, give full play to the clinical efficacy of HER2 targeted drug therapy, reduce treatment blindness, and benefit more patients. Member of the expert group of the breast cancer professional committee of China Anti-cancer Association, according to the "Expert Consensus on Clinical Diagnosis and Treatment of HER2-positive breast cancer" published in 2012^[1], combined with the research results at home and abroad, and updated as follows after expert discussion.

To analyze the types and distribution of large deletion of phenylalanine hydroxylase (PAH) gene in Chinese patients with phenylketonuria (PKU).

On the basis of 953 PKU patients from Peking Union Medical College and Gansu Province Medical Genetics Center, which were detected by directed sequencing of PAH gene between 2006 and 2014. Multiplex ligation-dependent probe amplification (MLPA) of PAH gene was performed in 43 patients with one or two unknown genotypes. And the deletion breakpoints were characterized by Gap PCR-sequencing.

Twenty-four large deletion/duplication alleles were found in 22 patients, accounting for 51.1%(24/47)of the 47 unknown mutations of the 43 patients.There were 6 different large deletions, including Ex1del3758 (n=10), Ex4_5del (n=4), Ex4_7del (n=3), Ex1del5329ins56 (n=3), Ex3del6599ins8 (n=2), and Ex4del (n=1); and 1 duplication was found (Ex12dup, n=1). The most common large deletions in Chinese patients were Ex1del3758 (21.3%), Ex4_5del (8.5%), and Ex4_7del (6.4%).

Large deletion mutations of PAH gene are present in Chinese PKU patients. It's important to detect the large del/dup mutation, and there are different hotspot mutation genotypes in Chinese patients.

To explore the role of microRNA-155 (miR-155) in pathogenesis of immune thrombocytopenia (ITP) through investigate the relevance between the expression of miR-155 in CD19⁺ B cells in peripheral blood and the function of B lymphocytes in patients with ITP.

A total of 55 ITP patients (30 newly diagnosed and 25 in remission) were collected from December 2014 to October 2015 in Tianjin Medical University General Hospital, and 25 healthy volunteers were recruited as controls. The CD19⁺ B cells were extracted by immunomagnetic microbeads. The expression of miR-155 in CD19⁺ B cells were detected by real-time fluorescent quantitative PCR. The levels of IgG, IgM and SH2 domain-containing inositol 5'-phosphatase 1 (SHIP-1) in CD19⁺ B cells were measured by flow cytometry (FCM). Correlation between miR-155 and clinical parameters of ITP patients was analyzed.

(1) The expression of miR-155 in CD19⁺ B cells in newly diagnosed ITP patients (4.57±1.03) was significantly higher than that in remitted ITP patients (0.79±0.13) and controls (1.74±0.32), and that in the remitted ITP patients was lower than in the controls (all P<0.05). In addition, the level of miR-155 at initial diagnosis in the patients responding to treatment (3.85±0.71) was lower than that in the refractory patients (6.67±1.05) (P<0.05). (2) The levels of IgG and IgM in CD19⁺ B cells in newly diagnosed ITP patients (35.20%±6.19%, 26.87%±5.01%) were significantly higher than those in remitted ITP patients (7.51%±1.91%, 5.93%±2.23%) and controls (8.23%±0.68%, 8.21%±1.08%) (all P<0.05). (3) The expression of SHIP-1 in CD19⁺ B cells in newly diagnosed ITP patients (44.33%±3.95%) was significantly lower than that in remitted ITP patients (83.13%±3.24%) and controls (67.26%±3.73%), and that in the remitted ITP patients was higher than in controls (all P<0.05). (4) The expression level of miR-155 in CD19⁺ B cells in ITP patients was positively correlated with CD19⁺ CD5⁺ B cell count (r=0.576, P<0.05), and negatively correlated with the level of SHIP-1 and peripheral platelet count (r=-0.445, -0.402, all P<0.05).

There is abnormally high expression of miR-155 in CD19⁺ B cells in peripheral blood of ITP patients, which is related with the dysfunction of B lymphocytes and ntracellular antibody production, suggesting that miR-155 might be involved in the pathogenesis of ITP.

To investigate the effects of melanin disappearance in vitiliginous skin on the ultrastructure of epidermal calcium distribution and the stratum corneum (SC) lipid lamellar membranes.

Five outpatients with stable vitiligo vulgaris and 5 healthy controls were recruited from August 2014 to February 2015 at Department of Dermatology, Renmin Hospital of Wuhan University. The ultrastructural changes of lipid lamellar membranes of the skin were assessed using transmission electron microscopy (TEM) technique in combination with ruthenium tetroxide (RuO₄) staining. The concentration and distribution of calcium precipitates in the epidermis were studied using calcium ion-capture cytochemistry combined with TEM.

The multilayered lipid lamellae existed within the intercellular space of the normal SC with a characteristic alternating electron-dense and electron-lucent pattern. Expanded intercellular space, fragmentation and lamellar separation were observed in the depigmented skin lesions from vitiligo patients, the bulbous regions of lipid lamellae were filled with electron-dense amorphous materials. Large clumps of calcium precipitates were visualized in the stratum granulosum (SG) of normal skin, fine calcium precipitates and stage Ⅳ melanosomes were noted within the normal stratum basale (SB). In depigmented skin lesions of vitiligo, both the size and number of calcium precipitates in the SG were dramatically decreased. Melanosome was barely seen in the vitiligo SB.

Disrupted ultrastructure of SC lamellar membranes and disappearance of calcium gradient co-exist in the skin lesion of vitiligo, indicating that melanin in epidermis may play a role in formation of epidermal calcium gradient and maintenance of structural integrity of permeability barrier.

To explore the risk factors of venous thromboembolism (VTE) recurrence and the predictive value of simplified pulmonary embolism severity index (sPESI) in medical inpatients.

A total of 149 consecutive patients with first diagnosed VTE from the medical departments of West China Hospital of Sichuan University from January 2011 and December 2012 were enrolled and followed-up for 24 months. The VTE recurrence rate was calculated and univariate and multivariate cox proportional hazards regression analysis were performed to identify the risk factors associated with VTE recurrence. All the patients were evaluated by sPESI, and survival analysis was used to explore its value in predicting VTE recurrence in these medical patients.

Out of the included 149 patients, 23(15.4%) patients had VTE recurrence during the 2 years' follow-up and median recurrence time was 167 days. The univariate analysis showed bed rest, severe lung disease, nephrotic syndrome, inappropriate anticoagulant therapy, smoking, diabetes, and malignant neoplasm might be associated with VTE recurrence (P=0.043, 0.006, 0.009, 0.032, 0.098, 0.048, 0.021). Among these risk factors, the multivariate analysis revealed severe lung disease, nephrotic syndrome, and malignant neoplasm were the independent risk factors (HR=3.45, 5.67, 3.60; P=0.020, 0.020, 0.047); while for inappropriate anticoagulant therapy, the P value was marginal (HR=3.94, 95% CI: 0.99-15.63, P=0.051). The median sPESI scores of the patients with VTE recurrence was higher than that of the patients without VTE recurrence[1(1, 2) vs 0(0, 1), P=0.001], and patients with sPESI≥1 were associated with 5.57-fold increased risk of VTE recurrence compared with patients with sPESI=0 (95%CI: 1.79-17.30, P=0.001). Survival analysis also showed that the 2-year cumulative VTE recurrence rate of patients with sPESI≥1 was significant higher than that of patients with sPESI=0 (38.4% vs 5.7%, P=0.001).

The medical VTE patients have high VTE recurrence risk, and severe lung disease, nephrotic syndrome, malignant neoplasm and inappropriate anticoagulant therapy are important risk factors of VTE recurrence. The sPESI has predictive value for VTE recurrence in medical patients.

To establish a multiplex polymerase chain reaction (mPCR) method with high sensitivity and specificity for rapid detection of common nontuberculous mycobacterium(NTM) infection in the hand.

Application of primer design software to the mycobacterium marinum, mycobacterium avium, mycobacterium kansasii and mycobacterium fortuitum, the specific gene sequences were used to design construction of multiplex PCR and detection of DNA from the non tuberculous mycobacterial standard strains of each bacterium of single PCR and multiplex DNA accuracy and sequence contrast evaluation to verify the specificity of multiple PCR primers.26 clinical specimens were identified by this method.

Detection of 26 cases of clinical samples, positive detection of more than 7/8, the identification time is shorter than the traditional method.

The research method can be rapid, specific, sensitive and effective to detect the common hand of mycobacterium tuberculosis infection, can be used for clinical identification of mycobacterium tuberculosis infection.

To quantify the expression of elafin mRNA in peripheral blood in patients with inflammatory bowel disease (IBD) and to explore its value in assessment of the activity and severity of IBD.

From July 1 2015 to August 15 2015, 23 patients with IBD admitted to Peking University First Hospital were selected, including 15 cases with ulcerative colitis (UC) and 8 cases with Crohn's disease (CD). Among those, 5 cases were in remission (UC 3, CD 2), 6 cases were mild active (UC 3, CD 3), 3 cases were moderate active (UC 1, CD 2), and 9 cases were severe active (UC 8, CD 1). A total of 21 healthy individuals were selected as the control group. Peripheral blood samples of IBD patients and healthy controls were collected. The expression of elafin mRNA in peripheral blood leukocytes was detected by fluorescence quantitative real-time PCR. Mann-Whitney test was performed for comparison between the two groups. The correlation between the expression of elafin mRNA in peripheral blood and IBD activity score was analyzed by Pearson correlation analysis after transformation of variables.

The median expression of elafin mRNA in peripheral blood leukocytes in IBD group and control group was 0.005 8 (0.000 2, 0.043 5) and 0.015 3 (0.002 1, 0.175 8), respectively, with no significant difference (P>0.05). However, in the active IBD patients it was lower than that in the controls (0.004 6 (0.000 2, 0.034 8) vs 0.015 3 (0.002 1, 0.175 8),P<0.05) and also lower than that in the remission patients(0.004 6 (0.000 2, 0.034 8) vs 0.023 1 (0.012 6, 0.043 5),P<0.05); in the active UC patients it was lower than that in the controls(0.003 7 (0.000 2, 0.027 0) vs 0.015 3 (0.002 1, 0.175 8),P<0.05). The expression of elafin mRNA in peripheral blood was negatively correlated with modified Mayo score in UC patients (r=-0.513, P<0.05) and with the Crohn's Disease Activity Index (CDAI) of Best score in CD patients (r=-0.889, P<0.05).

The expression of elafin mRNA in peripheral blood in active IBD patients is decreased, which may be correlated with the activity of IBD, and negatively correlated with corresponding disease activity score, suggesting that it may play a protective role in IBD and may be helpful in predicting disease activity.

To investigate the predictors of residual tumors at repeat transurethral resection of bladder tumors (re-TURBT) for the patients with T1 bladder cancer and evaluate the effect of the residual tumors on the prognosis of the disease.

We reviewed the clinical data of the patients with T1 bladder cancer who underwent re-TURBT from 2008 to 2015 in our department. Seventy-two patients received re-TURBT 2-6 weeks after the initial TURBT. A total of 65 patients were followed up, and we recorded the events of tumor recurrence, tumor progression, radical cystectomy and cancer specific death.The influencing factors of re-TURBT positive rate were analyzed.The effects of re-TURBT positive or negative findings on the prognosiswere compared.

33.3% of the patients who received re-TURBT had residual tumours. Re-TURBT positive in T1 bladder cancer has significant correlation with tumor size (P<0.05). Residual tumors tended to be detected in patients with larger tumors (diameter ≥ 4 cm) but might have no relationship with tumor grade or tumor number. The recurrence rate within 3 month ofthe patients with residual tumours at re-TURBT was 25% (5/20), while there were no patients suffering recurrence who had no residual tumours at re-TURBT, which had a significant difference (P<0.001). However, the overall recurrence rate, progression rate, rate of radical cystectomy and cancer specific mortality showed no significant difference between the two groups (45% vs 40%,P=0.71; 10% vs 6.7%, P=0.64; 5% vs 8.9%, P=0.59; 5% vs 2.2%, P=0.55).

For the patients with T1 bladder cancer, larger tumors could be a predictor for residual tumors at re-TURBT. The presence of residual tumors is associated with early recurrence.

To evaluate the clinical value of a new device designed by ourselves for removing the disposable circumcision device (Shang ring).

There were 264 patients undergoing Shang ring circumcision from August 2013 to June 2015. A total of 7 cases were lost follow-up and were excluded. The new device was applied for removing Shang ring in 150 patients after operation in Experimental group, and that was not applied in 107 cases in Control group. Values of basic demographic data, time for removing Shang ring, pain score, scar satisfaction, surgical satisfaction and complications were recorded and compared.

Time for removing Shang ring and pain score in Experimental group and Control group was 61.8±9.9 vs 127.4±41.6 s, 4.1±1.3 vs 6.8±1.4, respectively (both P<0.05). Satisfaction of surgery, percentage of willing to recommend Shang ring surgery to others in Experiment group and Control group was 9.3±0.7 vs 7.5±1.3, and 95.3% vs 79.4%, respectively (bothP<0.05). Surgical satisfaction was correlated with pain score, time of removing Shang ring and appearance of incision scar (P<0.05).

The new device for removing the Shang ring could remove the ring rapidly, alleviate pain and improve surgical satisfaction.

To evaluate the feasibility of retrograde transcatheter closure of ventricular septal defects (VSD) under trans-esophageal echocardiography (TEE).

Twelve patients with VSD, with 4 residual shunt, treated by transcatheter VSD closure under solely guidance of TEE were summarized. Mean age, body weight and average diameter of VSD in 12 cases were (11.58±6.57) years, (31.85±13.28) kg, (5.50±2.20) mm, respectively. All patients were treated by retrograde transcatheter closure under solely guidance of TEE. The post-operative efficiency was estimated by TEE immediately. Patients were followed up by echocardiography, X-ray examination and electrocardiogram at 24 hours, 1, 3 and 6 months after the procedures.

All 12 patients were treated successfully under solely guidance of TEE. The diameter of devices were 5-14 mm. There were 1 case with trivial residual shunt after the procedures immediately. No patients suffered from residual shunt, occluder malposition, peripheral vascular injury and pericardial effusion during the follow-up.

Percutaneous transcatheter closure of VSD completely guided by TEE is safe and effective. This method could avoid the radiation and contrast media reaction, and also could simplify the procedure.

To investigate the inhibitory effects of codonopsis pilosula polysaccharides (CPP) on the deterioration of impaired phagocytosis of alveolar macrophage (AM) induced by fine particulate matter with a mean aerodynamic diameter ≤2.5 μm (PM2.5) in chronic obstructive pulmonary disease (COPD) mice.

Sixty BALB/c male mice were randomly divided into control group, COPD group, PM2.5 group, PM2.5 COPD group, CPP COPD group and CPP+ PM2.5 COPD group. COPD mice were established using exposure of cigarette smoking. Meanwhile PM2.5 group, PM2.5 COPD group and CPP+ PM2.5 COPD group were exposed to PM2.5 (770 μg/m ³) for 90 days. CPP COPD group and CPP+ PM2.5 COPD group were fed with CPP (300 mg/kg) for 90 days whilst other groups were fed with isovolumetric saline. After the models were established, mice peak inspiratory flow (PIF) and peak expiratory flow (PEF) were measured by noninvasive body plethysmograph and lung histopathology and mean linear intercept (MLI) were observed. AMs were isolated from lung tissue by discontinuous density gradient centrifugation. Mean fluorescence intensity (MFI) and the ability of AM phagocytosing flurescein isothiocyanate-labeled Escherichia coli (FITC-E.coli) (AM%) were detected by flow cytometry. Total antioxidative capacity (TAC) was measured by O-phenanthroline colorimetry. Malondialdehyde (MDA) was measured by thiobarbiturieacid colorimetry and glutathione peroxidase (GSH-PX) by improved Hafeman colorimetry.

MFI in control group, COPD group, PM2.5 group, PM2.5 COPD group, CPP COPD group and CPP+ PM2.5 COPD group were 10 267±1 358, 4 817±399, 8 469±240, 3 176±501, 5 886±516 and 4 067±453. AM% in each group were (69.0±5.4)%, (30.7±3.0)%, (51.5±2.4)%, (20.4±3.5)%, (38.7±2.6)% and (28.7±4.3)%. MFI and AM% in COPD group and PM2.5 group were decreased than those in control group while those in PM2.5 COPD group were lower than in COPD group (all P<0.01). Comparing to COPD group and PM2.5 COPD group respectively, MFI and AM% in CPP COPD group and CPP + PM2.5 COPD group were increased (allP<0.01). TAC and GSH-PX in each group were (17.99±0.09), (6.83±0.36), (13.84±1.12), (3.61±0.29), (8.80±0.26), (5.43±0.30) U/mg protein and (84.3±5.7), (46.5±2.6), (62.0±2.2), (32.4±3.8), (53.4±4.0), (42.4±4.0) U/mg. TAC and GSH-PX in COPD group and PM2.5 group were lower than those from control group while those from PM2.5 COPD group were decreased than in COPD group (allP<0.01). Comparing to COPD group and PM2.5 COPD group, TAC and GSH-PX in CPP COPD group and CPP+ PM2.5 COPD group were increased respectively (allP<0.01). MDA in each group were (1.74±0.37), (2.73±0.22), (2.01±0.13), (3.55±0.33), (2.22±0.28) and (2.72±0.44) nmol/mg protein. MDA in COPD group and PM2.5 group were higher than that from control group while that from PM2.5 COPD group was higher than in COPD group; MDA in CPP COPD group and CPP+ PM2.5 COPD group were respectively decreased than those in COPD group and PM2.5 COPD group (allP<0.05). Positive correlations were existed between MFI, AM% and TAC, GSH-PX, while negative correlations were existed between MFI, AM% and MDA in COPD group, PM2.5 group, PM2.5 COPD group, CPP COPD group and CPP+ PM2.5 COPD group.

PM2.5 further impaired the defective phagocytosing capacity of AM and exacerbated oxidative stress in COPD mice. CPP can inhibit these effects. The protection of CPP may be closely related to its antioxidative effects.

To evaluate the effect of CD40 on Foxp3⁺ Treg cell in the lung of cigarette smoke exposure mice.

According to the random number table, 20 wild type (WT) C57 BL/6 mice and 20 CD40^-/-C57 BL/6 mice were randomly divided into two groups: WT control group, WT smoke-exposure group (24 weeks) and CD40^-/- control group, CD40^-/- smoke-exposure group (24 weeks) (n=10 each). Alveolar airspace enlargement was observed by HE staining. Morphological change was evaluated by mean linear intercepts (MLI). Immunohistochemical method was used to detect the quantity of Foxp3⁺ cell in the lung. The mRNA expression of Foxp3 was measured by fluorescence quantitative real-time polymerase chain reaction (qRT-PCR). The protein level of Foxp3 was measured by Western blot. Interleukin (IL)-10 and IL-35 levels in the lung were tested by enzyme-linked immunosorbent assay (ELISA).

The MLI in CD40^-/- smoke-exposure group was significantly lower than the WT smoke-exposure group[(30.0±1.7) vs (37.3±3.7) μm], but higher than the CD40 ^-/- control group[(23.2±2.5) μm], WT smoke-exposure group was significantly higher than the WT control group[(22.2±1.7) μm](all P<0.05). The percentage of Foxp3⁺ cell in the lungs of CD40^-/- smoke-exposure group was significantly higher than the WT smoke-exposure group and CD40^-/- control group[(16.89±0.75)% vs (9.65±0.74)% and (13.58±0.51)%], WT smoke-exposure group was significantly lower than WT control group[(12.13±0.81)%](allP<0.05). In the lungs, Foxp3 mRNA and protein expression in CD40^-/- smoke-exposure group were increased compared to WT-smoke-exposure group and CD40^-/- control group, WT smoke-exposure group were decreased compared to WT control group (all P<0.05). In the lungs, the level of IL-10 in CD40^-/- smoke-exposure group was higher than the WT smoke-exposure group and CD40^-/- control group[(231±25) vs (80±31) and (183±29) ng/L], WT smoke-exposure group was lower than the WT control group[(192±37) ng/L](allP<0.05). The level of IL-35 in CD40^-/- smoke-exposure group was higher than the CD40^-/- control group, WT control group and WT smoke-exposure group[(208±29) vs (118±29) , (148±36), (137±37) ng/L, all P<0.05].

Knockout the CD40 gene can promote the differentiation of Foxp3⁺ Treg cell in the lung of cigarette smoke exposure mice, indicating that blocking the CD40-CD40 ligand pathway may contribute to alleviate the smoking-induced pulmonary emphysema.

A 62-year-old male was admitted with high fever for 7 d. The patient had fever after catching cold, with a maximum body temperature of 41℃, accompanied by dry cough and fatigue. After intravenous treatment with etilmicin for 1 d, cephalosporins for 3 d and meropenem for 1 d in another hospital, his body temperature did not drop, and he developed transient delirium. He was admitted to our hospital with "community-acquired pneumonia". Previous diabetes for 7 years. Physical examination: body temperature 39.0 ℃, pulse 101 beats/min, blood pressure 140/80 mmHg (1 mmHg =0.133 kPa), 18 breaths/min. Sleepiness, a little wet rales can be heard in the left lung, but no obvious abnormalities are seen. Auxiliary examination blood routine: white blood cells 5.7×10⁹/L, neutrophils 75.7%; Urine routine: occult blood + +, protein + + +, glucose + +, 1.15 white blood cells/high power field; Blood gas analysis : pH 7.5, PaO₂65 mmHg, PaCO₂30 mmHg, HCO₃^-23.4 mmol/L; Blood CRP 125 mg/L, alanine aminotransferase 107 U/L, aspartate aminotransferase 105 U/L, blood glucose 9.46 mmol/L, potassium 3.74 mmol/L, sodium 126 mmol/L, D-dimer 977 μ g/L, cardiac enzyme, troponin I, renal function and blood amylase were not abnormal; Mycoplasma antibody, Legionella antibody, Legionella urine antigen and blood bacteria culture were all negative. Lung CT showed scattered consolidation in large ground glass shadows in the upper lobe of the left lung (FIG. 1A)。 After admission, the patient was given intravenous anti-infection and symptomatic supportive treatment with moxifloxacin, and the body temperature dropped to normal 72 hours after admission. Moxifloxacin was administered intravenously for 8 days and sequentially orally for 4 days. All indexes returned to normal after re-examination, and the pulmonary lesions were obviously absorbed compared with before (FIG. 1B), meeting the discharge criteria for community-acquired pneumonia (CAP). Treatment was not continued after discharge, and the body temperature rose again to 37.6 ℃ after 3 days. The results of Legionella antibody detection (indirect immunofluorescence method-German EU reagent) in the serum sample of the patient before discharge showed that the antibody of Legionella mickedei was positive with a titer of 1:320, and the patient was diagnosed as Legionella mickedei pneumonia. After stopping the drug, the patient's condition recurred, and he was instructed to continue taking moxifloxacin orally, and his body temperature dropped again. The overall course of treatment reached 3 weeks, and there was no recurrence.

The incidence of inflammatory bowel disease (IBD) has been increasing gradually in the past 50 years. Crohn's disease (CD) and ulcerative colitis (UC), as the two most common IBDs, have complicated clinical manifestations, low cure rate and easy recurrence and disability, which seriously affect the quality of life. It is agreed that the immune response to microbial abnormalities in the gut of genetically susceptible hosts is associated with IBD pathogenesis under the influence of environmental factors. A large number of literatures have reported that bacteria play an important role in the pathogenesis of IBD^[1]However, due to the relatively small number of fungi in the digestive tract and the limitation of experimental techniques, they have not received enough attention in previous studies. In recent 10 years, the development of high-throughput sequencing technology and bioinformatics has made it possible to explore the relationship between digestive tract fungi and IBD.

Allogeneic composite tissue transplantation is undoubtedly an ideal method for repair and reconstruction surgery to repair large area and special tissue defects (limb, face, penis, etc.). However, long-term high-dose immunosuppressive therapy after transplantation often leads to complications such as infection, diabetes, and even malignant tumor, which becomes an obstacle to the clinical development of allogeneic composite tissue transplantation^[1,2,3]。 Therefore, it is an urgent topic to explore the methods of inducing immune tolerance to reduce the dosage of immunosuppressant^[3]。 In recent years, many studies have shown that mesenchymal stem cells (MSCs) have immunomodulatory effects in animals in vitro and in vivo^[4,5]Some immunosuppressive effects were also observed with MSCs after clinical allotransplantation^[6]。