Volume 94  ·   Issue 48  ·  2014

Journal of Organ Transplantation (Transplantation) On April 27, 2014, Volume 97, Issue 8 published an article by Delmonico and eight other professors to our leaders in the form of an open letter: "China's Anti-Corruption Struggle in the Field of Organ Transplantation". Therefore, we express our views and positions on the issues involved in organ transplantation in China.

To observe the in vitro killing functions of natural killer (NK) cells for K562 cells in peripheral blood of severe aplastic anemia (SAA) patients and explore the role of NK cells in the immunological pathogenesis of SAA.

NK cells (CD3^-CD56⁺CD16⁺) as effector cells were sorted from peripheral blood mononuclear cells (PBMNC) of 12 SAA patients and 10 normal controls by magnetic activated cell sorting system (MACS)from June 2013 to August 2014. K562 cells as target cells were co-cultured with the effector cells (1∶1, 1∶5, 1∶10). Using Annexin V and PI double staining markers, the apoptotic rate of K562 cells were measured by flow cytometry after culturing.

The apoptotic rate of K562 cells in SAA patients was (10.30%±6.08%) according to the ratio of effector-target (1∶1). And it was significantly higher than that in normal controls (6.30%±3.46%) (P<0.05). The apoptotic rate of K562 cells in SAA patients were 10.30%±6.08%, 16.47%±8.29% and 25.45%±9.88% respectively according to the ratio of effector-target(1∶1, 5∶1, 10∶1). The higher ratio of effector-target, the higher apoptotic rate of K562 cells. There were statistic differences among three groups (P<0.05).

There are enhanced in vitro functions of NK cells in peripheral blood of SAA patients NK cells may play an important role in abnormal immune tolerance mechanisms of SAA patients.

To explore the levels of circulating hypoxia inducible factor-1α (HIF-1α) and hemeoxygenase-1 (HO-1) in liver cirrhosis and to explore their diagnostic values as noninvasive methods.

The levels of circulating HIF-1α and HO-1 were quantitatively detected in 34 patients with liver cirrhosis and 10 healthy controls by ELISA from May 2012 to May 2013. The diagnostic values were analyzed by ROC curve and their correlation with clinicopathological characters were also compared.

The serum levels of HIF-1α and HO-1 were significantly higher than those in healthy controls ((10.99±0.24) vs(5.79±0.84)μg/L, (63.04±1.87) vs (16.35±2.07) μg/L, bothP<0.01). No significant difference existed among Child-Pugh classification. The expressions of HIF-1α and HO-1 had a linear correlation between each other, and closely related with rebleeding and portal thrombosis during a one-year follow-up (allP<0.05). The area under ROC curve of HIF-1α and HO-1 was 0.894 and 0.994 respectively. With 9.45 μg/L as a critical point for HIF-1α, the diagnosis of liver cirrhosis might be predicted with a positive predictive value of 91.2%. And HO-1 >31.86 μg/L predicted the diagnosis of liver cirrhosis with a sensitivity of 97.1% and a specificity of 100%.

The serum levels of HIF-1α and HO-1 may serve as useful molecular markers for a noninvasive diagnosis of liver cirrhosis.

To explore the role of detecting the methylation status of gene Septin9 (SEPT9) in plasma for colorectal cancer screening in Chinese population.

Patients were collected from Beijing Military General Hospital since September 2013 to February 2014. The performance of SEPT9 assay was validated in a single-blind study of 80 cases with colonoscopy and pathologically verified colorectal cancer and 52 normal controls. The detection of Septin9 gene methylation in peripheral blood was performed by fluorescence quantitative polymerase chain reaction (PCR). And immunoassay fecal occult blood test was conducted to compare the superiority of methylated Septin9 for screening colorectal cancer.

The Septin9 assay successfully identified 75.0%(95%CI: 64.7%-83.6%) of cancers at a specificity of 98.1%(95%CI: 90.9%-99.9%). And it was superior to fecal occult blood screening for colorectal cancer (sensitivity 79.5% vs 53.8%, P<0.05).

Determination of SEPT9 methylation status is an innovative non-invasive plasma screening test for colorectal cancer.

To explore the causes of early failure after primary total hip arthroplasty (THA).

The causes of early THA failures were retrospectively reviewed for 215 cases of revision THA between January 2007 and December 2013.

The causes of early failures of primary THA in decreasing frequencies were infection (50.7%, 109/215), aseptic loosening (31.6%, 68/215), dislocation (8.8%, 19/215), periprosthetic fracture (3.7%, 8/215), implant-related failure (2.3%, 5/215), heterotopic ossification (1.4%, 3/215) and liner wear (0.9%, 2/215).

The top three causes of early failure after primary THA are infection, aseptic loosening and dislocation.

To explore the incidence of four types of chronic pain, i. e. headache, abdominal pain, neck & shoulder pain (NSP) and low back pain (LBP) and examine the relationship between the incidence of chronic pain and academic pressure in high school students.

A total of 3 000 high school students were randomly surveyed with a questionnaire on related issues. And the results were analyzed with a multivariate Logistic regression model.

Among them, 2 849 completed the questionnaire. And the overall incident rates of headache, abdominal pain, NSP, and LBP were 30.3%, 20.9%, 32.8% and 41.1% respectively. The students generally experienced a heavy burden of learning, a high level of stress and sleep deprivation closely related to four types of chronic pain.

As a common condition in Chinese adolescents, chronic pain is closely correlated with academic pressure.

To compare the clinical efficacies of mini-invasive percutaneous osteosynthesis (MIPO) versus supercutaneous plating with closed reduction in the treatment of distal tibial fractures.

A total of 48 patients with close distal tibial fractures were treated between January 2010 and January 2012. The MIPO group included 16 males and 8 females with an average age of 36 years. And the types were A (n=15), B (n=6) and C (n=3) according to the classification scheme of Association for the Study of Internal Fixation (AO/ASIF). The supercutaneous plating group also included 16 males and 8 females with an average age of 37 years. And the types were A (n=15), B (n=6) and C (n=3). And the operative duration, hospital stay, union time, postoperative complications and function of ankle were compared between two groups.

The mean follow-up period was 18.5 (12-26) months. There was no instance of nonunion, hardware breakdown or deep infection. Patients in supercutaneous plating group had significantly shorter mean operative duration, hospital stay and union time. Three patients and 1 patient in MIPO group presented with superficial infection and delayed union respectively while there was no occurrence in supercutaneous plating group. And the differences were not statistically significant. Fifteen patients (62.5%) complained of implant impingement or discomfort. And stripping occurred at an incidence of 15.6% during the removal time of locking screws in MIPO group. While in supercutaneous plating group, there as no complaint of skin irritation and removal of supercutaneous plate was easily performed without anesthesia. The mean AOFAS score was 90.7±3.8 in supercutaneous plating group versus 88.9±4.1 in MIPO group (P=0.070).

Distal tibia fractures may be treated successfully with MIPO or supercutaneous plating. However, supercutaneous plating offers multiple advantages in terms of mean operative duration, hospital stay, union time, skin irritation and implant removal.

To evaluate and compare the radiographic parameters between patients of adult acquired flatfoot and normal participants to provide rationales for diagnosis and operative reconstruction.

The adult acquired deformity group consisted of 85 patients (85 feet) and normal group 90 normal participants (90 feet) from January 2008 to May 2014.The weight-bearing anteroposterior and lateral foot radiographs were taken.And the following parameters were measured and compared: medial column length (MCL), lateral column length (LCL), medial column height (MCH), lateral column height (LCH), talonavicular uncoverage angle (TNUA), talus-first metatarsal angle (TMTA) and calcaneal pitch angle (CPA). Age and all parameters were included for Logistic regression analysis.

On anteroposterior radiographs, the values of MCL for adult acquired deformity and normal groups were (103.5±9.0), (105.8±7.8)mm and LCL (94.6±6.9), (96.2±6.1)mm respectively.No significant difference existed in MCL or LCL (P>0.05). However, significant inter-group differences existed in TMTA and TNUA (P<0.01). The values of TMTA were (18.9±2.5)°, (7.8±2.2)° and TNUA (23.6±3.2)°, (8.0±2.2)°.On lateral radiographs, significant inter-group differences existed in MCH, LCH, CPA and TMTA (P<0.01). And the values of MCH were (16.3±1.7), (23.6±2.8)mm, LCH (14.9±1.1), (13.96±1.39)mm, CPA (14.86±2.76)°, (22.9±1.9)° and TMTA (10.0±2.0)°, (-3.6±1.0)° respectively.According to Logistic regression analysis, the significance of parameters ranged from lateral TMTA (B=0.032), anteroposterior TNUA (B=0.014), anteroposterior TMTA (B=0.015) to lateral MCH (B=-0.012), lateral CPA B (B=-0.009).

No differences exist in anteroposterior MCL or LCL.There are positive differences in lateral TMTA, anteroposterior TNUA and anteroposterior TMTA.And negative differences exist in lateral MCH and lateral CPA between adult flatfoot and normal foot.Lateral TMTA may be an excellent radiographic identifier of patients with adult acquired flatfoot during diagnosis and treatment.

To explore the differential expression profiles and transcriptomes of human bone microvascular endothelial cells via microarray and elucidate the molecular mechanisms underlying steroids-induced osteonecrosis.

Bone microvascular endothelial cells in cancellous bone of femoral head were harvested by type Ⅰ collagenase and trypsin digestion and identified by immunofluorescent labeling with primary antibodies specific for vascular endothelial cells' biomarkers such as von Willebrand factor, platelet endothelial cell adhesion molecule 1, vascular endothelial cadherin and vascular cell adhesion molecule 1. Their differential expression profiles and transcriptomes were tested by double-labeled lncRNAs + mRNAs microarrays and single-labeledd microRNAs microarrays. The data were analyzed by bioinformatical software with database checking. The authors spatial correlations and expression relevance were established among lncRNAs, microRNAs and mRNAs.

Steroids incurred specific changes in mRNAs, microRNAs and lncRNAs transcribed in human bone microvascular endothelial cells of femoral head in vitro cultured. Thus a model of coexpression network was constructed among transcript modules with bioinformatical tools such as target gene prediction.

Steroids induced significant changes in transcripts of bone microvascular endothelial cells such as miR-100, miR-222, miR-933, miR-339 and miR-23. They exert considerable influences on osseous tissues and circulatory systems in cancellous bone by targeting cytokines and enzymes such catalase, fibroblast growth factor and nerve growth factor. Through further explorations of these transcripts, we can not only elucidate the underlying pathogenesis, but also discover specific biomarkers and therapeutic targets for corresponding diseases.

To explore the effect of pectin, a kind of soluble dietary fiber, on colonic transit time (CTT), clinical symptoms and gut microbiota in adults with slow-transit constipation.

A total of 80 patients with slow-transit constipation were selected between June 2011 and December 2013. For this randomized controlled trial, we evaluated the efficacy of pectin on intestinal transit time and other indices of constipation in adults with slow-transit constipation. They were randomized to receive either pectin or placebo. Treatment consisted of 4-week supplementation with 24 g/d pectin (fiber group) or maltodextrin (placebo group). Before and after 4-week treatment, CTT, constipation symptoms and fecal bacterial population were compared between groups.

CTT of the fiber group after treatment was lower than those of fiber group before treatment and those in the placebo group ((60.2±11.2) h vs (80.3±9.5), (79.4±11.7) h, P<0.01). Constipation score of the fiber group after treatment also decreased than those of fiber group before treatment and those in the placebo group (bothP<0.05).Bifidobacterium sp. and Lactobacillus sp. evidently increased (lg copies/g: 8.26±0.83 vs 6.42±1.07 and 6.48±0.82, 6.83±0.77 vs 5.85±0.64 and 5.91±0.73; all P<0.05)and totalClostridium sp. significantly decreased (9.07±0.63 vs 9.74±0.81 and 9.66±0.43, P<0.05) in the fiber group after treatment versus the placebo group. No significant adverse effects were reported.

Four-week soluble dietary fiber use accelerates colon transit time and alleviates clinical symptoms in patients with slow-transit constipation. Additionally, supplementary fiber offers protective effects on gut microbiota by increasing the population of healthy microflora.

To explore the role of P70S6 kinase phosphorylation as a downstream of mammalian target of rapamycin (mTOR) pathway in CD3 positive cells of liver transplant patients.

A total of 84 liver transplant recipients were recruited from our hospital and divided into 3 treatment groups of sirolimus (n=26), tacrolimus (n=35) and cyclosporine (n=23). The P70S6 kinase phosphorylation of CD3 positive cell of patients and healthy control (HC) were analyzed by phospho-flow cytometry. A correlation analysis between P70S6 kinase phosphorylation and sirolimus trough level was performed. Intra-individual variability and inter-individual variability of P70S6 kinase phosphorylation were measured.

The P70S6 kinase phosphorylation in HC showed a low degree of intra-individual variability (3.5% to 5.6%) while the inter-individual variability between different healthy volunteers was higher (18.9% to 22.5%). The P70S6 kinase phosphorylation in patients treated with sirolimus (28.9±10.5) was significantly lower than in HC (57.2±8.4, P<0.001), tacrolimus (42.5±14.1,P<0.001) or cyclosporine treated ones (51.4±10.9,P<0.001). The P70S6 kinase phosphorylation in HC (57.2±8.4)was significantly higher than in tacrolimus (42.5±14.1,P<0.01) or cyclosporine treated patients (51.4±10.9,P<0.05). No correlation existed between P70S6 kinase phosphorylation and trough level of sirolimus (r=-0.18, P=0.39).

Phospho-flow cytometry assay has been established for determining the degree of mTOR inhibition by assessing P70S6 kinase phosphorylation. Quantification of P70S6 kinase phosphorylation may play an adjunct role in pharmacodynamically guiding an individualized mTOR inhibitor based immunosuppression.

To explore the effects of membrane skeleton protein 4.1R on the efficiency of photodynamic therapy (PDT).

4.1R gene knockout and wild-type mouse embryonic fibroblasts (MEFs) were incubated with various concentrations of 5-aminolevulinic acid (5-ALA) (0.25, 0.50, 1.00, 1.50 and 2.00 mmol/L), followed by exposure to 450 nm light at a dose of 72, 96, 120, 180, 240 mJ/cm². Cell counting kit 8 (CCK-8) assay was used to assess the survival rate after PDT treatment. Laser confocal microscopy was used to observe the location of photo-sensitizer protoporphyrin and fluorescence spectrophotometer for detecting the fluorescent intensity of intracellular protoporphyrin. The protein levels of rate-limiting enzyme of protoporphyrin synthesis, ferrochelatase (FECH) and hydroxymethylbilane synthase (HMBS) were determined by Western blot.

Both cell lines were killed after 5-aminolevulinic acid (5-ALA)-PDT and its efficacy was dependent on 5-ALA concentration, incubation duration and light dose. The cell survival rates of 4.1R^-/- MEF were significantly higher than those of 4.1R^{+ /+}MEF (46.9%±7.1% vs 12.5%±2.1%, P<0.001) after PDT treatment with a light dose of 120 mJ/cm² mediated by 5-ALA 1.00 mmol/L. After incubation with 1.00 mmol/L 5-ALA, protoporphyrin was distributed throughout cytoplasm in both cell lines while the fluorescent intensity of 4.1R^{+ /+}MEF was higher than that of 4.1R^-/- MEF (124.2±3.5 vs 34.6±3.8, P<0.001). Western blot showed that no difference of FECH and HMBS protein level was found in two cell lines.

A lack of protein 4.1R may attenuate the intracellular protoporphyrin level and the photo-cytotoxicity of PDT. No cellular change of ALA metabolic activity is found. Protein 4.1R may be involved in the ALA uptake in MEF cells so that the cellular level of protoporphyrin ultimately affects the PDT efficiency.

To compare the efficacy of lapatinib or lapatinib plus trastuzumab versus trastuzumab in the neoadjuvant therapy of human epidermal growth factor receptor 2 (HER-2) positive breast cancer.

MEDLINE database, American Society of Clinical Oncology (ASCO), San Antonio Breast Cancer Symposium (SABCS), European Society for Medical Oncology (ESMO) proceedings and China Biomedical Database were searched for literatures of trastuzumab or lapatinib in neoadjuvant therapy for breast cancer. There was no limit of language or time. A meta-analysis was performed for retrieved literatures meeting the inclusion criteria.

A total of 1 794 breast cancer patients from 5 clinical trials were included. And the regimens were lapatinib plus neoadjuvant chemotherapy (n=719), trastuzumab plus neoadjuvant chemotherapy (n=714) and both drugs plus neoadjuvant chemotherapy (n=361). The rate of pathological complete remission (pCR) was lower in lapatinib group than that in trastuzumab group (28.2% vs 35.4%). And the difference was statistically significant (P=0.004). The pCR rate was significantly higher in lapatinib plus trastuzumab therapy group than that in trastuzumab group (53.2% vs 38.1%, P<0.001).

Lapatinib can not replace trastuzumab as a first-choice agent in neoadjuvant therapy of HER-2 positive breast cancer. Lapatinib plus trastuzumab achieves better pCR than trastuzumab so that it and may become a first-choice of neoadjuvant therapy for HER-2 positive breast cancer regardless of economic affordability for patients.

The patient, an 85-year-old male, was admitted to the hospital on April 2, 2014 due to intermittent epistaxis for more than 5 years, general ecchymosis for more than 4 years, and aggravation for more than 2 months. The patient had persistent epistaxis without obvious trigger since 2008, and finally stopped bleeding after tamponade and compression by otolaryngology. Since then, patients will have epistaxis 1 to 3 times almost every winter, and all of them need tamponade to stop bleeding. Since 2009, patients have intermittently developed post-bump skin ecchymosis, which is more common in the limbs, with different sizes and no obvious pain, and no diagnosis and treatment have been performed. In February 2014, the patient developed large ecchymosis on his left hand, about 4 cm in diameter, after bumping. Later, due to braces falling off and injury, blood blisters with a diameter of about 2 cm appeared on the tongue, and the pain was obvious, which affected eating. In the same month, there were patches of ecchymosis in the left popliteal fossa, posterior femur, hip and waist without obvious trigger. Routine blood test in other hospitals: white blood cell count normal, hemoglobin (HGB) 103 g/L, mean red blood cell volume (MCV) 95.4 fl, platelet 80×10⁹/L; Ferritin normal (345.2 μ g/L); Coagulation function: prothrombin time (PT) 13.5 s increased (↑), activated partial thromboplastin time (aPTT) 45.9 s ↑, thrombin time (TT) 20 s ↑; Liver and kidney function: creatinine 204 μ mol/L↑, total bilirubin (TBil) 33.3 μ mol/L↑, direct bilirubin (DBil) 20.4 μ mol/L↑, and symptomatic treatment for hemostasis. On March 25, 2014, the patient accidentally fell from bed and landed on his left hip. The next day, the left hip mass, swelling and pain appeared, and the skin gradually appeared bruised. Admitted to our hospital as "cause of bleeding tendency to be investigated". Since the onset of the disease, there was no fever, night sweats, fatigue, dizziness, amaurosis, palpitations, decreased activity tolerance, acceptable spirit, sleep and diet, normal defecation, and no obvious change in weight. Past history: "Pulmonary tuberculosis" was found in 1955 and recovered after treatment. Discovery of atrial fibrillation in 1975. In 2009, creatinine was found to be elevated (the details are unknown), and no diagnosis and treatment were performed. In 2011, he went to an outside hospital for "left pleural effusion and left lower lobe nodules". After thoracic puncture and drainage, the effusion absorption disappeared, and benign inflammatory lesions were considered. Discovery of hypertension in 2012. In 1989, he underwent surgery for "intestinal obstruction", the details of which are unknown. Family history: father died of "gastric cancer".

immune thrombocytopenic purpura (ITP) is a common hemorrhagic disorder characterized by the presence of antiplatelet antibodies in circulating peripheral blood, decreased platelet count and increased risk of bleeding. At present, it has been found that the activation and clearance of macrophages mediated by platelet-specific autoantibody Fc segment produced by B cells is one of the main pathogenesis of ITP^[1]。 B-cell activating factor (BAFF) is a newly discovered member of the tumor necrosis factor (TNF) superfamily, which can promote B lymphocyte survival, maintain germinal center response and T cell activation, and play an important role in autoimmune diseases.

Intraductal breast carcinoma (DCIS) is a breast carcinoma in situ that is confined to the breast ductal system and does not invade the basement membrane and surrounding stroma. Some patients can stay "in situ" for a long time, while others can progress to invasive carcinoma^[1]。 Since the 1980s, with the improvement of mammography technology and the popularization of application, the detection rate of DCIS has been greatly increased^[2,3]。 In the United States, for example, the incidence of DCIS increased from 5.8 per 100,000 person-years in 1975 to 32.5 per 100,000 person-years in 2004, accounting for about 20% of new breast cancer cases each year^[4]。 At present, there is a great controversy about the surgical treatment strategy of intraductal breast carcinoma. It is a difficult problem for surgeons to choose the appropriate surgical method that can not only prevent the development of the natural course of the disease, but also avoid the harm to patients caused by overtreatment. Therefore, this article intends to summarize the controversial issues and research progress in the field of surgical treatment.

Childhood obstructive sleep apnea hypopnea syndrome (OSAHS) refers to a series of pathophysiological changes caused by frequent partial or total upper airway obstruction during sleep, which disturbs children's normal sleep ventilation and sleep structure^[1]。 Its common clinical manifestations include snoring during sleep, mouth breathing, apnea, repeated waking-up, sweating, etc., which can also cause a series of complications, such as inattention, poor memory, mental impairment, low weight or obesity, growth retardation and non-specific behavior difficulties, and pulmonary heart disease or even death in severe cases. Among them, growth retardation has attracted the attention of more and more medical scholars. The research progress on the influence of OSAHS on children's height and its mechanism at home and abroad is summarized as follows.

A 22-year-old male was admitted to Tianjin Haihe Hospital on June 8, 2013 due to "fever, headache for half a month and confusion for 3 days". The patient developed fever and headache half a month ago, with a maximum body temperature of 40 ℃, accompanied by nausea and no vomiting. Anti-infective therapy was given in other hospitals, and intracranial pressure was measured by lumbar puncture up to 500 mmH₂O (1 mmH₂O =9.807×10^–3kPa), considered "viral meningitis", and the effect of antiviral and symptomatic treatment (specificity unknown) is not good. Three days ago, he developed disturbance of consciousness, agitation and coma, and successfully disconnected after tracheotomy and mechanical ventilation for three days. After consultation in our hospital, we considered "tuberculous meningitis" and transferred into the hospital.