Volume 95  ·  Issue 46  ·  2015

In recent years, targeted drugs such as epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) have become one of the important treatments for advanced non-small cell lung cancer (NSCLC). In patients with advanced NSCLC with EGFR gene mutation, a series of international and domestic multicenter clinical studies have confirmed that EGFR targeted therapy can significantly reduce the risk of disease progression or death and improve the quality of life of patients. In lung cancer patients without EGFR gene mutation, clinical research evidence suggests that targeted drugs should not be used. These clinical studies and systematic analyses indicate that EGFR gene mutation detection is a prerequisite for the use of EGFR-TKIs in patients with advanced lung cancer.

To research the suppressive effect of interferon α2b (IFN-α2b) on angiogenesis of JAK2 tyrosine kinase gene point mutation (JAK2V617F) positive myeloproliferative neoplasm (MPN) and its anti-angiogenic mechanisms.

(1) A total of 42 cases of JAK2V617F positive MPN patients in the No.1 Hospital of Baoding were selected between January 2012 and October 2014, including the newly diagnosed group before treatment (n=25) and the IFN-α2b treatment group (n=17). Ten cases of idiopathic immune thrombocytopenic purpura (ITP) patients were enrolled as controls. JAK2V617F/JAK2 ratio was detected by real-time fluorescent quantitative PCR to measure mutation; the expression levels of phosphorylated JAK2 (p-JAK2), vascular endothelial growth factor (VEGF), hypoxia-inducible factor-1α (HIF-1α) and CD105 marked microvascular density (MVD) in bone marrow were detected by immunohistochemistry. The correlations were analyzed among JAK2V617F mutation burden and VEGF, HIF-1α, and MVD. (2) Human erythroleukemia cell line HEL cells were treated with different concentrations of IFN-α2b. The apoptosis was detected by Hoechst staining method. The cell viability was detected by CCK-8 test. Cell migration ability was tested by transwell chambers. The protein expression levels of p-JAK2, VEGF, and HIF-1α were detected by Western blot.

(1)The ratio of JAK2V617F/JAK2 in the IFN-α2b treatment group (22.69%±12.64%) was significantly lower than that in the newly diagnosed group (46.17%±19.32%) (P<0.01). The expression levels of p-JAK2, VEGF, and HIF-1α in the newly diagnosed group (82.41%±11.65%, 64.72%±25.01%, 45.12%±20.28%) were significantly higher than those in the IFN-α2b treatment group (60.93%±20.57%, 36.58%±15.95%, 32.15%±14.27%) and the control group (43.05%±12.59%, 25.69%±13.75%, 25.07%±15.49%) (allP<0.01). MVD in the newly diagnosed group (26.58%±5.93%) was significantly higher than that in the treatment group (15.86%±4.27%) and the control group (10.76%±4.01%) (P<0.01). Spearman correlation analysis showed positive correlation of JAK2V617F mutation with VEGF and MVD (r=0.589, P<0.05;r=0.577, P<0.05). (2)The apoptosis of HEL cells were increased after treated with 30 000 U/L of IFN-α2b in HEL cells after 24 h. The proliferation of HEL cell were inhibited by different concentrations of IFN-α2b in time- and dose-dependent manner. The number of membrane-permeating HEL cells after treated with 5 000 U/L of IFN-α2b in HEL cells after 24 h was significantly lower than that in the untreated cells (52.9±7.5 vs 77.3±6.1) (P<0.05). The expression levels of p-JAK2, VEGF, and HIF-1α were decreased in a dose-dependent manner.

IFN-α2b may exert an anti-angiogenic effect by inhibiting the VEGF, HIF-1α, and MVD expression in MPNvia JAK2 signal pathway.

To assess the value of different kinds of cryptococcal capsular antigen detection methods in diagnosis and efficacy assessment in the patients with cryptococcal meningoencephalitis.

From October 2012 to March 2015, cerebrospinal fluid (CSF) samples were collected from 57 patients withcentral nervous system infection who hospitalized in Sir Run Run Shaw Hospital. The cryptococcal capsular antigen in CSF was detected by Lateral flow assay (LFA), Latex agglutination test(LA) and Enzyme immunoassay (EIA). Follow-up study was achieved in 10 patients by detecting the samples of their CSF using LA and EIA, so that the dynamic changes of the antigen level could be obtained and observed during the treatment.

The sensitivity of LFA and LA were both 95%, meanwhile the specificity were both 100%; the sensitivity and specificity of EIA were both 100%. The level of cryptococcal capsular polysaccharide antigen decreased gradually with patients recovering.

All the three different methods could beof great importance for the early diagnosis of cryptococcal meningoencephalitis. LFA should be recommended for screening the disease. Follow-up study by detecting the antigen level of CSF is valued in assessing the treatment efficacy of cryptococcal diseases.

To investigate the expression of miRNA-4497 in human chorionic villi from early recurrent miscarriage and the influence on apoptosis.

The expression of miRNA-4497 in chorionic villi tissues collected from 20 early recurrent miscarriage cases and 20 controls were analyzed using quantitative real-time polymerase chain reaction (qRT-PCR). Human BeWo trophoblastic strains transfected with miRNA-4497 mimic were collected before transfection and 24 h later to measure apoptosis by flow cytometry.

miRNA-4497 was up-regulated in early recurrent miscarriage (24.5%±5.5% vs 1.1%±0.1%, P<0.05), and there was significantly increase in apoptosis after miRNA-4497 mimic transfection (12.2%±2.2% vs 7.3%±1.2%,P<0.05).

Up-regulation of miR-4497 may play an important role in the pathogenesis of recurrent miscarriage. And the pathophysiological mechanism involved should be further studied.

To explore the reliability of the Geriatric Nutritional Risk Index (GNRI) as a mortality predictor in elderly patients undergoing hemodialysis (HD).

A total of 125 maintenance HD patients aged >60 years old who had received dialysis for over 6 months before entry was retrospectively examined. The values of GNRI were calculated, and death was taken as the end point. The patients were divided into 4 groups according to the quartiles of GNRI values. All-cause and cardiovascular mortality were calculated using Kaplan-Meier and cox proportional-hazards analyses, and ROC curve was adopted for analyzing the predicting value of GNRI on mortality.

The GNRI of the four groups were ≤92.06, 92.07-96.15, 96.16-101.25, ≥101.26, respectively. Kaplan-Meier survival rate was significantly different among 4 groups for all-cause and cardiovascular mortality. Cox regression model analysis demonstrated that the GNRI was a predictor for all cause (HR=0.940, P=0.001, 95%CI: 0.907-0.974) and cardiovascular mortality (HR=0.906, P<0.001, 95%CI: 0.863-0.951). The area under ROC curve was 0.667 (P=0.001) for all-cause mortality and 0.717 (P=0.001) for cardiovascular mortality.

The GNRI is a reliable predictor for all-cause mortality in maintainance HD patients, but more multi-center studies with larger samples are still needed.

To study the correlationship between the BRCA1 gene polymorphisms, especially in 2731 loci (rs799917), and sporadic breast cancer in the Han nationality and the Mongol nationality of the Inner Mongolia region.

Using the prospective study method, 103 cases of patients with sporadic breast cancer (case group) and 103 cases of normal physical examination people (control group) were enrolled. PCR and direct sequencing method were used for analyzing the correlationship of 2731 loci polymorphisms of BRCA1 and sporadic breast cancer in our zone.

In the case group, the age stratification, pathologic stage, immunohistochemistry and the distribution of lymph node metastasis had no significant difference in two ethnic group (P> 0.05). The age stratification of control group also had no significant difference in two ethnic group (P>0. 05). There was no statistically significant difference in age stratification of the case group and the control group (P>0.05). In the Inner Mongolia region, BRCA1 gene 2731 loci genotypes check out three genotypes: namely TT, CT and CC. The frequencies of genotype TT, CT, CC in the case group were 13.1%, 26.2%, 60.7% ( the Han nationality) and 16.7%, 28.6%, 54.7% (the Mongol nationality), respectively. Meanwhile the frequencies of allele T and allele C were 71.8% and 28.2%. In the control group, the frequencies of genotype TT, CT, CC were 18.0%, 31.1%, 50.9% ( the Han nationality) and 23.8%, 38.1%, 38.1% ( the Mongol nationality), respectively, and the frequencies of allele T and allele C were 62.9% and 37.1%. BRCA1 gene 2 731 loci gene polymorphism had no significant difference in two groups (χ²=3.438, P=0.752), but T allele frequency distribution in the case group was significantly increased (χ²=4.185, P=0.041).

There is no obvious correlation between the BRCA1 gene 2731 loci and sporadic breast cancer in the Han nationality and the Mongol nationality of the Inner Mongolia region. C allele of BRCA1 gene 2731 loci may be one of the protective gene of sporadic breast cancer.

To investigate the clinical presentation, histopathological features, progression, and treatment of lymphomatoid papulosis (LyP).

A retrospective review was performed on clinicopathological data of 22 patients diagnosed with LyP from June 2010 to March 2015 in Institute of Dermatology, Chinese Academy of Medical Sciences and Peking Union Medical College.

The mean age of the 22 LyP patients was 39 years (range: 7-83 years). The male-to-female ratio was 1∶1. The areas predominantly affected were the trunks, followed by limbs and face. Most lesions presented as recurrent eruption of erythema, papule, nodules, ulcer, necrosis and crusting. Some of them leaved hyperpigmentation or atrophic scars on healing. Histopathologically, LyP were devided into types A, B, C, D and E, with 12, 1, 1, 3, and 4 cases in each type, respectively, and one case of mixed type B and C. One of the patients was also diagnosed with primary cutaneous anaplastic large-cell lymphoma besides the diagnosis of LyP. Among the 15 patients with follow-up information available, 11 patients were treated with regimens including oral corticosteroids, methotrexate, tripchlorolide, intramuscular injection of interferon, phototherapy, and topical corticosteroids. The mean follow-up time was 22(1-54)months.All the patients were alive at the end of the follow-up period.

LyP is a low-grade malignant T-cell lymphoma with a benign clinical course but histologically malignant features. Multi-agent chemotherapy is unnecessary. Patients with LyP are likely to have an favorable prognosis.

To explore the effect of three-dimensional (3D) and two-dimensional (2D) acellular fibrotic lung matrices on fibroblast-to-myofibroblast differentiation.

Twenty adult SD rats were randomly divided into control group and idiopathic pulmonary fibrosis (IPF) group. The pulmonary fibrosis was induced by Bleomycin. Normal and fibrotic decellularized lungs were made, then sections with 500 μm (3D) and 10 μm (2D) thick were cut by a standard Vibratome. The lung fibroblasts were isolated from the lungs of SD rats at postnatal day 1, which were seeded dropwise into different slices (2D/3D normal and fibrotic scaffolds), then cultured for 72 hours in vitro. The protein expression of alpha smooth muscle actin (α-SMA) was measured by immunofluorescence staining and western blot.

There was no spatial structure both in 2D normal scaffold and fibrotic scaffold. The spatial structure was different between 3D normal scaffold and fibrotic scaffold. The relative expression amount of α-SMA in 2D normal and fibrotic scaffold group and none scaffold group were (4.64±0.36, 4.86±0.37, 4.67±0.35, respectively), there were no difference among them ( P>0.05). In 3D fibrotic scaffold group, the relative expression amount of α-SMA was significantly increased compared with those in 3D normal scaffold group and 2D fibrotic scaffold group (6.29±0.85 vs 4.85±0.56, 4.86±0.37, bothP<0.05).

3D acellular fibrotic lung scaffold can promote fibroblast-to-myofibroblast differentiation.

To explore the association between acute inflammatory reaction and hemodynamic changes of acute pulmonary embolism rabbit with hyperhomocysteinemia.

A total of 30 Japanese white rabbits were randomly divided into homocysteine group and control group according to the random number table, which were treated with 2% methionine and normal saline, respectively. Rabbit model of acute pulmonary embolism was established by autologous blood clots reinfusion after 8 weeks, then the level of serum tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β), right ventricular systolic pressure (RVSP), right atria systolic pressure (RASP), pulmonary artery pressure (PAP) were checked before and after thrombosis at different time points. At last, area of embolism, degree of lung congestion and incidence of lung infarction were evaluated after the rabbits were sacrificed.

Both homocysteine group and control group serum TNF-α, IL-1β increased at 30, 60, 120 min after embolism, and serum TNF-α, IL-1β of homocysteine group [(12.4±1.0), (15.7±1.1), (23.2±1.4) and (7.6±1.0), (10.2±2.1), (18.8±1.3) pg/ml] increased more obviously than control group [(10.2±1.0), (11.7±1.4), (12.1±1.2) and (6.4±1.1), (8.5±1.9), (10.0±2.1) pg/ml] (allP<0.05). All of RVSP, RASP, and PAP increased in each group after embolism, while homocysteine group RVSP [(52.8±3.3), (54.7±4.5), (46.4±4.4) mmHg], RASP [(3.9±1.2), (4.5±2.0), (4.5±1.9) mmHg] and PAP[(52.9±3.3), (55.1±2.9), (47.5±3.6) mmHg] increased more obviously compared with control group [(39.4±2.5), (39.4±3.3), (34.5±3.9) mmHg, (2.3±1.1), (3.3±1.1), (3.3±1.2) mmHg and (37.7±2.7), (40.2±2.9), (33.7±4.2) mmHg] (allP<0.05).

Inflammatory response of acute pulmonary embolism rabbit increases in hyperhomocysteinemia and the degree of embolism increases as well, which may lead to increase of hemodynamic instability.

To explore the expression of pulmonary surfactant in rats with paraquat-induced acute lung injury.

A total of 36 SD rats were randomly divided into the control group (n=6) and paraquat group at 6, 12, 24, 48, 72 h five time points, each time points 6 rats. All of the rats were injected with paraquat (80 mg/kg) for once. The specimens were collected at 0 h (instant) after paraquat intoxication in the control group, and at the corresponding point in time in the paraquat each time point groups. The pathology changes of lung tissue were observed by HE staining, thiobarbituric acid method was used to detect malondialdehyde (MDA) level, and Western blot method was used to analyze the relative expression of surfactant protein (SP)-A and SP-D in bronchoalveolar lavage fluid (BALF) with time.

As compared to control, the levels of MDA in BALF were significantly higher at 6, 24, 48 h after paraquat intoxication ((4.19±0.12), (3.33±0.08), (3.52±0.08) vs (2.82±0.15) nmol/mg, all P<0.01), no significantly different at 12, 72 h ((2.76±0.13), (2.79±0.10) nmol/mg, bothP>0.05); the relative expression of SP-A in BALF were significantly higher at 6 h after paraquat intoxication (1.32±0.19 vs 1.00±0.19,P<0.01), lower at 24, 48 h (0.43±0.07, 0.67±0.08, bothP<0.01), and no significantly different at 12, 72 h (0.98±0.15, 0.79±0.18, bothP>0.05); the relative expression of SP-D in BALF were significantly higher at 6, 12 h after paraquat intoxication (1.54±0.33, 1.64±0.37 vs 1.00±0.23, bothP<0.01), no significantly different at 24, 48 h (1.07±0.19, 0.97±0.15, bothP>0.05), and reached the peak at 72 h (2.15±0.26,P<0.01); the expression of MDA, SP-A and SP-D were characterized by the state of fluctuation with increased first, reduced after, and finally increased.

The expression of SP-A, SP-D increase at the early stage of the paraquat-induced acute lung injury in rat, which can reflect the degree of lung injury.

To investigate the expressions of Notch3, Notch4, Frizzled2 and Tead1 at protein levels in the cortex of frontal and parietal lobes after focal cerebral ischemia-reperfusion (I/R) in rats.

The focal cerebral I/R model was established by intraluminal thread occlusion of the middle cerebral artery (MCAO). The animals were divided into sham operation group (sham group) and I/R group. The infarct area of the brain was measured by 2, 3, 5-triphenyl tetrazolium chloride (TTC) staining technique after 1 week. Protein expression levels of Notch3 and Frizzled2 in the cortex of frontal and parietal lobes were detected by immunohistochemical staining, and protein expression levels of Notch4 and Tead1 were detected by Western blot analysis.

The infarct area of rats in I/R group was very obvious. The protein expressions of Notch3 and Frizzled2 in I/R group were higher than those in sham group (P<0.05,P<0.01). The protein expression of Notch4 in I/R group was significantly higher than that in sham group (P<0.01), but the protein expression level of Tead1 in I/R group was significantly lower than that in sham group(P<0.01).

Notch signal transduction pathway was activated after focal cerebral ischemia/reperfusion in MCAO rats.

To investigate the association of thyroid autoimmunity (TAI) with pregnancy outcomes in euthyroid women undergoing in vitro fertilization (IVF).

The meta-analysis was a systematic review of cohort studies published before May 10th, 2014. Data were retrieved from Medline, Embase and Cochrane library, and the keywords were"thyroid autoimmunity"and"pregnancy outcome", with restriction to artificial reproductive technique. RevMan 5.2 software was used for data analysis.

In all, 11 cohort studies met the inclusion criteria and were included in the meta-analysis. The results showed that TAI was not associated with clinical pregnancy rate (RR=0.99, 95%CI: 0.89-1.11, P=0.92), but significantly associated with miscarriage risk (RR=1.54, 95%CI: 1.24-1.92, P<0.001). Live birth/ongoing pregnancy rate decreased in the TAI group compared with the control group, but there was no statistically significant difference (RR=0.89, 95%CI: 0.79-1.00, P=0.06).

As for euthyroid women who are receiving IVF, TAI contributes to increased risk of miscarriage. Therefore, we recommend universal screening for thyroid disease among women undergoing IVF.

To verify the feasibility of human papilloma virus(HPV) subtypes detection by AUTRAX automatic nucleic acid extraction workstation.

A total of 183 HPV test samples (2 562 types) were collected during August 2014 in Peking Union Medical College Hospital. Nucleic acid determination kit of high-risk HPV types (16, 18, 35, 39, 58, 31, 33, 68, 56, 45, 59, 51, 52) and 6 , 11 type (real-time PCR) were applied for detection. Each sample was divided into two parts. One part was treated with manual extraction, which entailed manually preparing PCR reaction system and added the sample to the PCR plate. Another was treated with the AUTRAX automatic nucleic acid extraction workstation, which automatically prepared the reaction system and added to the PCR board. These two parts proceeded to the real-time PCR detection. The result of manual extraction was set as the golden standard and the Kappa consistency analysis was conducted. Meanwhile, precision and pollution prevention of the AUTRAX were verified.

The sensitivity of AUTRAX automatic nucleic acids extraction workstation was 97.12% (101/104). The specificity was 99.51% (2 446/2 458). The accuracy was 99.42% (2 547/2 562). The result of Kappa consistency analysis showed that the two parts were the same (Kappa=0.926). Coefficient of variation (CV) of each HPV types was less than 5%.No pollution phenomenon was found.

AUTRAX automatic nucleic acids extraction workstation can be used for the HPV subtypes detection in clinical settings.

Penicillium marneffei (PSM) is an invasive deep fungal disease caused by infection with Penicillium marneffei (PM)^[1]。 At present, the exact route of infection is still unknown, and it often causes disseminated infection in the whole body, even invades the main airway and bronchi, causing the destruction, collapse and obstruction of airway, resulting in fatal asphyxia^[2]。 Due to the rare reports of PM invasion of airway, routine imaging examination is difficult to detect, and diagnosis is difficult. Misdiagnosis and misdiagnosis often occur, and eventually serious complications occur. From January 2013 to June 2015, three patients with PM diagnosed and treated in the Department of Respiratory Medicine, the First Affiliated Hospital of Guangxi Medical University were diagnosed with disseminated PSM with tracheal-bronchial-pulmonary involvement due to repeated lung shadow treatment. The etiological basis was finally obtained by fiberoptic bronchoscopy (fiberoptic bronchoscopy). Satisfactory results were achieved after aerosol inhalation and combined intravenous application of amphotericin B.

A 38-year-old female was admitted to the hospital on 28 April 2014 due to "intermittent upper abdominal discomfort for 2 years and abdominal mass found for 2 months". Two years ago, the patient experienced paroxysmal upper abdominal discomfort with abdominal pain and hiccups, which occurred 15 to 16 times a month. It appeared 10 to 15 minutes after eating, and relieved spontaneously after about 1 to 2 hours, without headache, palpitations and hyperhidrosis. He had a history of constipation for 2 years, and the patient was not diagnosed and treated. The patient had no abdominal pain, anorexia, nausea, vomiting, no melena and weight loss, no erythema migrans, no chest stiffness, shortness of breath, good spirit and appetite, dry stool for the past 4 to 5 years, and was given glycerol preparations to assist in defecation. On 9 March 2014, the tumor marker CA199 was 55.08 U/ml (normal<37.00 U/ml). Abdominal B-ultrasound: About 116 mm ×65 mm mixed echoes were seen in the right abdomen, with clear boundaries, and most of them were solid echoes, showing multiple anechoic areas. Abdominal CT: A mixed density mass shadow was seen in the abdominal cavity, about 96 mm ×66 mm in size, compressing the inferior vena cava, and the gastric wall in the gastric antrum was slightly thickened. Heterogeneous enhancement of the mass after enhancement. Consider abdominal space-occupying lesions, pancreatic space-occupying lesions, excluding stromal tumors (Figure 1)。 Admitted to the pancreatic group of basic surgery, and completed the relevant examinations: complete blood cell analysis: red blood cell count 4.15×10¹²/L, hemoglobin 99 g/L, hematocrit (HCT) 0.322. Urinary routine: 100 red blood cells/HP, gastroscopy: chronic superficial gastritis, external pressure bulge of descending duodenum (Figure 2)。 Helicobacter pylori (HP) negative (–). Endoscopic ultrasound: cystic mass of external pressure in the descending duodenum with normal pancreatic echo (Figure 3)。 Pancreatic thin scan enhanced CT: A massive soft tissue shadow, about 81 mm ×129 mm, was seen in the right upper abdomen, with uneven density, and multiple patchy cystic necrosis was seen inside. Considering the right upper abdominal mass, gastrointestinal stromal tumor is more likely (Figure 4)。 The tumor was mainly supplied by the branch of the abdominal aorta. According to the blood supply of the abdominal aorta and the location of the tumor, paraganglioma was not excluded. It was transferred to the urology department on March 26. Qualitative examination: urinary catecholamines (CA): norepinephrine (NE) 132.3 μ g/24 h, epinephrine (E) 6.3 μ g/24 h, dopamine (DA) 7 975.8 μ g/24 h (normal reference range: 120.9~330.6 μ g/24 h), all of which were significantly increased; Localization examination: Interadrenal medullary iodobenzguanidine imaging (MIBG) suggested an area equivalent to the right side of the middle abdomen with increased radioactivity, considered paraganglioma, with internal necrosis. Octreotide imaging showed high expression of somatostatin receptor in the right side of the middle abdomen, which considered the possibility of neuroendocrine tumor. Abdominal ultrasound on 31 Mar 2014: A 12.8 cm ×10.0 cm ×7.2 cm hypoechoic mass was seen on the right side of the abdominal aorta, in front of the renal vessels and the inferior vena cava in the right abdomen.

The patient was a 66-year-old male. He was admitted to hospital on March 3, 2015 due to "low back pain and discomfort for 10 days, headache, fever with gibberish for 5 days, and coma for 1 day". Lower back pain, accompanied by fatigue, no fever, no painful urination and urgency occurred without obvious trigger 10 days ago, which was not paid attention to. Fever occurred about 5 days ago, body temperature 37.7~38.6 ℃, accompanied by headache, bilateral temporal distension and pain, no cough and sputum, no vomiting, no convulsions, occasional gibberish, can quickly return to normal by itself, visited the local hospital, blood routine showed white blood cells 8.4×10⁹/L, the percentage of neutrophils was 0.92, C-reactive protein: 160 mg/L, the head CT showed "lacunar infarction in left parieto-occipital lobe", and the abdomen CT showed "liver abscess". There was no significant improvement after anti-inflammatory administration of "aloxicillin sodium". 1 d ago, the patient had headache, gibberish, irritability was worse than before, and then he became comatose, so he was transferred to the emergency department of our hospital. Since the onset of the disease, urine and defecation as usual, loss of appetite, and general sleep. He had a history of diabetes for 2 years and was not treated with regular medication. Denial of medical history such as hypertension. Denial of biliary stones or history of surgery. A family genetic history is denied. History of allergies denied.

Diabetes is a chronic syndrome caused by hyperglycemia, which affects the whole body and involves the nervous system, which is called diabetic neuropathy. Neuropathy may affect all somatic nerves, among which peripheral neuropathy, which is mainly distal limb paresthesia, is the most common. Neuralgia can occur in 6% of patients with diabetic peripheral neuropathy (DPN)^[1]painful diabetic neuropathy (PDN) is characterized by cautery-like, knife-like, insect bite-like, tear-like, and needle-like pain at night. The etiology and pathogenesis of PDN are still unknown, and it is generally believed that its occurrence is related to a combination of factors. In recent years, the role of inflammation in PDN has attracted more and more attention. The pro-inflammatory cytokine tumor necrosis factor α (TNF-α) is highly expressed in the serum of patients with type 1 diabetes and type 2 diabetes, and the expression of TNF-α in DPN patients is significantly increased compared with that of patients with simple diabetes. The higher the level of TNF-α in patients with DPN, the more likely it is to complicate with painful neuropathy, and the higher the level of TNF-α in patients' serum, the more severe the pain response. TNF-α may be involved in the occurrence of diabetic painful neuropathy, leading to nerve ischemia, local nerve fiber demyelination, pain and heat hyperesthesia, etc. PDN care is complicated, and repeated hospitalization brings huge economic losses, which reduces the quality of life of patients, and can lead to non-traumatic amputation, high disability rate and mortality rate. At present, the only drugs approved by the US Food and Drug Regulatory Authority and the European Medicines Agency for the treatment of diabetic painful neuropathy are duloxetine and pregabalin^[2]。 Through the study of TNF-α, it is possible to find new potential therapeutic targets for diabetic painful neuropathy.

Aplastic anemia (AA) includes congenital and acquired, the former being Fanconi anemia; The latter is an acquired myelohematopoietic failure, characterized by myelohematopoietic failure, pancytopenia, and high mortality, but the exact etiology and mechanism remain unclarified. Recent studies have shown that its pathogenesis is related to the hyperfunction of autoimmune T cells. Autoreactive pathogenic T cells expand and activate, polarize to helper T cells (Th) 1 response and secrete a large number of inflammatory cytokines, directly or indirectly damage hematopoietic stem progenitor cells and bone marrow microenvironment. It is a class of T lymphocyte-mediated autoimmune diseases targeting hematopoietic system^[1,2]。

Lymphoproliferative diseases such as acute and chronic lymphocytic leukemia, lymphoma, multiple myeloma, etc. are difficult to diagnose. Among them, non-Hodgkin's lymphoma is the most complicated and confusing type of disease in pathological classification, and non-Hodgkin's lymphoma accounts for 89% of malignant lymphomas in China^[1]。 Techniques for diagnosis, efficacy and prognosis of non-Hodgkin's lymphoma mainly include conventional methods such as morphology, immunohistochemistry and cytogenetics. Immunophenotype analysis, including flow cytometry and immunohistochemistry, is the best auxiliary tool for the diagnosis of lymphoma. It plays a decisive role in the analysis of lymphoma subtypes, and also plays an auxiliary role in the judgment of lymphocyte clonal proliferation^[2]。 However, many lymphomas lack clonal proliferation markers, so immunophenotyping methods relying on these markers have certain limitations. It has been reported that approximately 5% to 10% of lymphoma cases are still difficult to obtain a final diagnosis after multiple phenotypic analyses^[3]At this time, it is necessary to rely on a molecular diagnostic technology, that is, the clonality analysis of receptor gene rearrangement to make a judgment^[4]。 Receptor gene rearrangement clonality analysis has been used in the diagnosis of lymphoma for nearly 30 years. Southern blot hybridization was initially used, but this method has strict requirements for specimen quality, large amount of tissue and complicated operation, so its clinical application is limited. Since 1990s, PCR has been used in the clonality analysis of recipient gene rearrangement and has gradually replaced Southern blot hybridization^[5,6]。 In recent years, researchers have improved the detection technology of recipient gene rearrangement mainly focusing on improving primers and establishing simpler and more accurate product analysis methods.