Objective To investigate the optimized parameters of dye (SYBR Green Ⅰ) realtime fluorescent polymerase chain reaction (RF-PCR) for detecting αβT lymphocyte clones in the peripheral blood and its application in monitoring specific T cell clone in the peripheral blood mononuclear cells (PBMC) from patients with chronic hepatitis B (CHB). Methods The total RNA was extracted from the PBMC of six healthy donors, and was reversely transcripted into cDNA. Then the cDNA was amplified using RF-PCR with the primers specific for T cell receptor β viable region (TCRBV) gene families as upstream primers and the primer for T cell receptor (TCR) β constant region (TCRBC) as downstream primer. The annealing temperature,concentration of primers and the total number of cycles were comparatively analyzed. The optimized PCR was performed to investigate the 24 TCRBV gene families from 12 patients with CHB, and the PCR products were monitored by melting curve analysis, and the clone expansion of peripheral blood T cell was detected by peak-motif of melting curve analysis. Results The optimized annealing temperature, final premier concentration,the number of cycles were 60.6 ℃, 0.5 μmol/L and 40 cycles, respectively. The begin temperature for melting curve analysis was better as 80 ℃ compared to 75 ℃. There was mono-peak on melting peak chart for TCRBV gene families in PBMC from patients with CHB, and PCR products of the single peak were determined as monoclonal T cell by sequencing. Conclusions The optimized reaction parameters of RF-PCR for monitoring 24 TCRBV gene families are determined. The melting peak chart could be used to monitor the clone expansion of the peripheral lymphocytes and to detect the clone-specific T cells in the peripheral blood from patients with CHB.

Objective To summarize the experiences in the diagnosis and treatment of the hepatic hereditary hemorrhagic telangiectasia (HHHT).Methods The clinical data of 15 HHHT patients who were admitted to the Qilu Hospital,People's Hospital of Mengyin,People's Hospital of Liaocheng,Henan Provincial People's Hospital,the Second Hospital of Hebei Medical University,First Affiliated Hospital of Zhejiang University were retrospectively analyzed.The clinical manifestation,features of imaging and laboratory examination were summarized,and the diagnosis,treatment and prognosis of the disease were investigated.Results HHHT patients had nonspecific symptoms in the early stage,and some patients presented with right upper quadrant discomfort,shortness of breath,anemia and liver bruit.The condition of HHHT patients could be worsened by liver cirrhosis or portal hypertension rapidly.The results of color doppler ultrasound and computed tomography showed intrahepatic telangiectasia,arteriovenous fistula and hepatic artery aneurysm in the 15 patients.Digital subtraction angiography was not clear enough for 2 HHHT patients with more than 1 enlarged hepatic arteries,but computed tomographic angiography was feasible.According to the degree and stages of the HHHT,all the 15 patients were divided into asymptomatic HHHT,simple HHHT and complex HHHT.Among the 6 patients who underwent surgical treatment,5 received ligation or banding of the enlarged hepatic arteries with subsequent disappearance of symptoms.Three patients received interventional treatment,and the treatment for 1 patient with complex HHHT was failed,and the patient died 30 months after medical treatment.Six patients were treated by conservative treatment,2 patients of them had no symptoms at the beginning,then they suffered from hepatic dysfunction and ascites at 21 and 35 months,respectively,and 1 of them died 6 months later.Four patients received medical treatment,and the results of color doppler ultrasound and computed tomography showed the pathological changes were aggravated gradually.Conclusions Telangiectasia,intrahepatic arteriovenous fistula and hepatic artery aneurysm are the main imaging characteristics of HHHT,and imaging diagnosis has significant value in the diagnosis of HHHT.HHHT is a progressive disease,early,active and individualized treatment is beneficial to the patients.The outcome of ligation or banding of the hepatic arteries is satisfactory.

Objective To investigate JC virus(JCV) infection in kidney transplant recipients and its influence on graft function and also initially explore JCV infection factors. Methods A total of 49 kidney transplant recipients and 24 health examination persons were enrolled in our study, JCV DNA was measured using nested qualitative polymerase chain reaction assays of urine, while CMV DNA was measured by common qualitative polymerase chain reaction assays of urine. JCV infection factors, such as age, male, immunosuppressive therapy, cytomegalovirus(CMV) infection were analyzed by Binary Logistic Regression, and glomerular filtration rate(GFR) was selected as a index of kidney function and the difference of GFR between JCV-infected and non-infected patients was compared using t test. Results JCV was detected in 42.9% of kidney transplant patients and 4.2% health examination persons. CMV infection and Pred + MMF + CsA triple immunosuppressive regimen were found to be the risk factors of JCV infection. No difference of GFR was observed between JCV infected and non-infected patients (86.470 ± 29.990 and 84.060 ± 33. 729 for each; t =0. 259, P =0.797). Conclusion JCV is frequently detected in kidney transplant recipients. CMV infection and using of Pred + MMF + CsA triple immunosuppressive regimen can significantly increase the risk of JCV infection. While, graft function was not influenced by JCV infection in kidney transplant patients.

Objective: To study serum HCV antibody (anti-HCV) with geographic distribution characteristics in Zhejiang Province. Methods: A stratified random cluster sampling method was used. Serum samples of the surveyed population were collected from selected hospitals, anti-HCV antibodies were examined, then hepatitis C infection rates among different genders, regions and age groups were analyzed. The anti-HCV rate was compared using the χ ² test. Results: The average anti-HCV positive rate in Zhejiang Province was 0.24% [95% confidence interval (CI): 0.16% ~ 0.32%]. The antibody positive rate in the plain area was 0.32% (95% CI: 0.19% ~ 0.45%), which was significantly higher than the coastal islands 0.05%(95% CI: 0.00% ~ 0.12%, χ ² = 7.638, P < 0.05). There was no significant difference between plain area and hilly area 0.22% (95% CI: 0.03% - 0.41%). There was no statistically significant difference in anti-HCV positive rates between males and females (χ ² = 2.238, P = 0.135). The highest positive rate of anti-HCV (0.93%) was in the population aged 56-60 years and the lowest in the population aged less than 20 years. Anti-HCV positive rate of all age groups in 2017 was lower than that of 2006 seroepidemiological study of hepatitis C. Conclusion Zhejiang Province is a region with low anti-HCV positive rate and the disease prevalence further reduced than 10 years ago. The positive rate of anti-HCV in plain areas is higher than islands. Middle-aged and elderly people are the age group with high prevalence, and the anti-HCV positive rate in people under 20 years old is exceptionally low. Gender differences in anti-HCV positive rate have little effect.