Prediction of the Impact of CYP2C19 Polymorphism on Drug-Drug Interaction between Voriconazole and Tacrolimus Using Physiologically-Based Pharmacokinetic Modelling

Abstract Voriconazole increases tacrolimus blood concentration significantly when coadministrated. The recommendation of reducing tacrolimus to 1/3 in voriconazole package insert seems not to be satisfactory in clinical practice. In vitro studies demonstrated that the magnitude of inhibition depends on the concentration of voriconazole, while voriconazole exposure is determined by the genotype status of CYP2C19. CYP2C19 gene polymorphism challenges the management of drug-drug interactions(DDIs) between voriconazole and tacrolimus. This work aimed to predict the impact of CYP2C19 polymorphism on the DDIs by using physiologically based pharmacokinetics (PBPK) models. The precision of the developed voriconazole and tacrolimus models was reasonable by evaluating the pharmacokinetic parameters fold error, such as AUC0-24, Cmax and tmax. Voriconazole increased tacrolimus concentration immediately in all population. The simulated duration of DDIs disappearance after voriconazole withdrawal were 146h, 90h and 66h in poor metabolizers (PMs), intermediate metabolizers (IMs) and extensive metabolizers(EMs), respectively. The developed and optimized PBPK models in this study can be applied to assit the dose adjustment for tacrolimus with and without voriconazole.

The epidemic trend of viral hepatitis in Nanjing from 1989 to 2020 / 预防医学

Objective@#To analyze the epidemic trend of viral hepatitis in Nanjing from 1989 to 2019 and predict the incidence in 2020, so as to provide reference for the prevention and control of viral hepatitis.@*Methods@#The incidence data of viral hepatitis in Nanjing from 1989 to 2019 was retrieved from Nanjng Center for Disease Control and Prevention and National Infectious Disease Reporting System. The epidemic trend was analyzed by estimating the annual percent change ( APC ) and the average annual percent change ( AAPC ). The seasonal incidence of different types of viral hepatitis was analyzed by seasonal index. The autoregressive integrated moving average model ( ARIMA ) was built to predict monthly incidence rate of viral hepatitis in 2020. @*Results@#The annual incidence rate of viral hepatitis was 62.00/100 000 in Nanjing from 1989 to 2019, showing a downward trend ( AAPC=8.4%, P<0.05 ). From 1998 to 2019, the annual incidence rates of hepatitis A, B, C and E were 1.98/100 000, 14.31/100 000, 2.30/100 000 and 2.60/100 000. The incidence of hepatitis A and B showed downward trends ( AAPC=-11.81%, -6.02%, both P<0.05 ); the incidence trend of hepatitis C was not obvious ( P>0.05 ); the incidence of hepatitis E showed an increasing trend ( AAPC=4.82%, P<0.05 ). From 2015 to 2019, the third and fourth quarters were the epidemic seasons of hepatitis A, B and C, while the first and second quarters were the epidemic seasons of hepatitis E. The ARIMA model predicted that the monthly incidence rates of viral hepatitis in 2020 would range from 1.26/100 000 to 3.69/100 000, among which hepatitis B ranged from 1.21/100 000 to 2.58/100 000, hepatitis C from 0.20/100 000 to 0.48/100 000, hepatitis E from 0.09/100 000 to 0.25/100 000. @*Conclusions@#The incidence of viral hepatitis in Nanjing shows a downward trend. Among different types of hepatitis, hepatitis B has a higher incidence. All types of hepatitis have epidemic seasons. It is predicted that the monthly incidence rates of viral hepatitis will be 1.26/100 000 to 3.69/100 000 in 2020.

Effect on Treatment of Addition and Subtraction Therapy of Zhuyu Zhixuetang to Chronic Endometritis with Syndrome of Qi Deficiency and Blood Stasis / 中国实验方剂学杂志

Objective:To discuss clinical effect of Zhuyu Zhixuetang to chronic endometritis （CE） with syndrome of Qi deficiency and blood stasis and to study improvement effect to pregnancy outcome. Method:One hundred and forty-four patients were randomly divided into two groups， patients in control group were 72 cases and in observation group were 72 cases. In control group， 66 patients completed the therapy because of 4 falling off or missing visit and 2 eliminated， and in observation group， 65 patients completed the therapy because of 3 falling off or missing visit and 5 eliminated. In two groups. anti-infection treatment was gicen to patients. Patients in control group got Fuke Qianjin Pian， 6 tablets/time， 3 times/day. Patients in observation group got Zhuyu Zhixuetang， 1 dose/day. The treatment was continued for 3 months and the follow up was recorded for 6 months. Before and after treatment， changes of menstrual volume， period and cycle were recorded. And hysteroscopy and color Doppler ultrasound of vagina were made， and endometrial morphology and endometrial receptivity were evaluated ［endometrial thickness， resistance index （RI）， pulsation index （PI） and blood flow index （FI）］ were evaluated， and pathology of endometrial were tested. And scores of syndrome of Qi deficiency and blood stasis were graded， levels of interleukin-1 <italic>β</italic> （IL-1<italic> β</italic>）， IL-6 and tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>） and T-lymphoid subsets （CD3⁺， CD4⁺， CD8⁺） in peripheral blood were measured. Pregnancy and miscarriage were recorded and the safety was evaluated. Result:After treatment， menstrual volume， menstrual period， cycle and complete normal rate of menstruation in observation group were all higher than those in control group （<italic>P</italic><0.05）. Endometrial thickness and FI were more than those in control group （<italic>P</italic><0.01）， RI and PI were lower than RI and PI in control group （<italic>P</italic><0.01）. And compared with control group， levels of IL-1<italic>β</italic>， IL-6， TNF-<italic>α</italic> and CD8⁺were less （<italic>P</italic><0.01）. And levels of CD3⁺， CD4⁺ and CD4⁺/ CD8⁺ were higher than the data in control group （<italic>P</italic><0.01）. During 6 months follow-up， pregnancy rate in observation group was 46.97% （31/66） was higher than 27.69% （18/65） in control group （<italic>χ</italic>²=5.197， <italic>P</italic><0.05）. Total effective rate of endometrial morphology was 96.97% （64/66） higher than 86.15% （56/65） in control group （<italic>χ</italic>²=4.981， <italic>P</italic><0.05）. Total effective rate endometrial pathology was 95.45% （63/66） higher than 84.62% （55/65） in control group （<italic>χ</italic>²=4.304， <italic>P</italic><0.05）. Total effective rate of comprehensive clinical effect was 93.94% （62/66） higher than 81.54% （55/65） in control group （<italic>χ</italic>²=4.696， <italic>P</italic><0.05）. There was no adverse reactions related to traditional Chinese medicine. Conclusion:Zhuyu Zhixuetang can regulate menstruation， relieve clinical symptoms， improve endometrial morphology under hysteroscopy， regulate systemic and local immune inflammatory response， improve CP， thus improve pregnancy outcome， with better comprehensive effect and safety.

MicroRNA-143–5p modulates pulmonary artery smooth muscle cells functions in hypoxic pulmonary hypertension through targeting HIF-1α

This paper explores the potential mechanism of microRNA-143–5p regulation effects on pulmonary arterysmooth muscle cells (PASMCs) functions in hypoxic pulmonary hypertension (HPH) via targeting HIF-1a,which may offer a new idea for HPH therapy. PASMCs were transfected with mimics control/miR-143–5pmimics or inhibitor control/miR-143–5p inhibitor. We used Western blotting and RT-qPCR to detect the proteinand mRNA expressions, CCK-8 assay to detect cellular viability, Annexin V-FITC/PI staining and caspase3/cleaved caspase-3 protein to evaluate cellular apoptosis, transwell migration experiment for cellularmigration measurement and Dual luciferase reporter gene assay to prove the target of miR-143–5p. Cells underhypoxic condition presented the decreased protein and mRNA expressions of a-smooth muscle actin (SM-aactin), Myocardin, smooth muscle myosin heavy chain (SMMHC), and smooth muscle-22a (SM22a),Calponin1 and Hypoxia-inducible factor-1a(HIF-1a), the increased cell viability and miR-143–5p level; Overexpression of miR-143–5p obviously reduced vascular smooth muscle-specific contraction marker proteinlevels and cellular apoptosis, increased cellular migration of PASMCs with hypoxia stimulation; Low-expression of miR-143–5p caused the opposite changes, while co-transfected with Si HIF-1a blocked thebeneficial effects of miR-143–5p inhibition on PASMCs under hypoxia. MicroRNA-143–5p can promote thephenotype conversion, proliferation and migration of pulmonary artery smooth muscle cells under hypoxiccondition through direct targeting of HIF-1a.

Effects of Rab7 gene up-regulation on renal fibrosis induced by unilateral ureteral obstruction

Rab7, an important member of the Rab family, is closely related to autophagy, endocytosis, apoptosis, and tumor suppression but few studies have described its association with renal fibrosis. In the early stage, our group studied the effects of Rab7 on production and degradation of extracellular matrix in hypoxic renal tubular epithelial cells. Because cell culture in vitro is different from the environment in vivo, it is urgent to understand the effects in vivo. In our current study, we established a renal fibrosis model in Rab7-knock-in mice (prepared by CRISPR/Cas9 technology) and wild type (WT) C57BL/6 mice using unilateral ureteral obstruction (UUO). Seven and 14 days after UUO, the expression of the Rab7 protein in WT mice, as well as the autophagic activity, renal function, and the degree of renal fibrosis in WT and Rab7-knock-in mice were examined by blood biochemical assay, hematoxylin-eosin and Masson staining, immunohistochemistry, and western blotting. We found that the Rab7 expression in WT mice increased over time. Furthermore, the autophagic activity constantly increased in both groups, although it was higher in the Rab7-knock-in mice than in the WT mice at the same time point. Seven days after UUO, the degree of renal fibrosis was milder in the Rab7-knock-in mice than in the WT mice, but it became more severe 14 days after surgery. Similar results were found for renal function. Therefore, Rab7 suppressed renal fibrosis in mice initially, but eventually it aggravated fibrosis with the activation of autophagy.

Unsatisfying antiviral therapeutic effect in patients with mother-tochild transmissed chronic hepatitis B virus infection: a prospective multi-center clinical study / 中华医学杂志(英文版)

Background@#Few data are available regarding the progression of liver disease and therapeutic efficacy in chronic hepatitis B virus (HBV) carriers infected by mother-to-child transmission (MTCT). This study aimed to investigate these two aspects by comparing the adult chronic HBV carriers in MTCT group with those in horizontal transmission group.@*Methods@#The 683 adult chronic HBV patients qualified for liver biopsy including 191 with MTCT and 492 with horizontal transmission entered the multi-center prospective study from October 2013 to May 2016. Biopsy results from 217 patients at baseline and 78 weeks post antiviral therapy were collected.@*Results@#Patients infected by MTCT were more likely to have e antigen positive (68.6% vs. 58.2%, χ2 = -2.491, P = 0.012) than those with horizontal transmission. However, in patients with MTCT, levels of alkaline phosphatase (ALP) (P = 0.031), Fibroscan (P = 0.013), N-terminal propeptide of Type III procollagen (PIIINP) (P = 0.014), and Laminin (LN) (P = 0.006) were high, in contrast to the patients with horizontal transmission for whom the levels of albumin (ALB) (P = 0.041), matrix metalloproteinase-3 (MMP-3) (P = 0.001) were high. The 47.2% of patients with MTCT and 36.8% of those with horizontal transmission had significant liver fibrosis (P = 0.013). Following antiviral therapy for 78 weeks, 21.2% and 38.0% patients with MTCT and horizontal transmission acquired hepatitis B e antigen (HBeAg) clearance, respectively (P = 0.043), and the virological response rates were 54.7% and 74.1% in the MTCT and horizontal groups, respectively (P = 0.005). MTCT was a risk factor for HBeAg clearance and virological response.@*Conclusion@#Adult patients with MTCT were more prone to severe liver diseases, and the therapeutic efficacy was relatively poor, which underlined the importance of earlier, long-term treatment and interrupting perinatal transmission.@*Trial Registration@#NCT01962155; https://clinicaltrials.gov.

Unsatisfying antiviral therapeutic effect in patients with mother-to-child transmissed chronic hepatitis B virus infection: a prospective multi-center clinical study / 中华医学杂志(英文版)

BACKGROUND@#Few data are available regarding the progression of liver disease and therapeutic efficacy in chronic hepatitis B virus (HBV) carriers infected by mother-to-child transmission (MTCT). This study aimed to investigate these two aspects by comparing the adult chronic HBV carriers in MTCT group with those in horizontal transmission group.@*METHODS@#The 683 adult chronic HBV patients qualified for liver biopsy including 191 with MTCT and 492 with horizontal transmission entered the multi-center prospective study from October 2013 to May 2016. Biopsy results from 217 patients at baseline and 78 weeks post antiviral therapy were collected.@*RESULTS@#Patients infected by MTCT were more likely to have e antigen positive (68.6% vs. 58.2%, χ = -2.491, P = 0.012) than those with horizontal transmission. However, in patients with MTCT, levels of alkaline phosphatase (ALP) (P = 0.031), Fibroscan (P = 0.013), N-terminal propeptide of Type III procollagen (PIIINP) (P = 0.014), and Laminin (LN) (P = 0.006) were high, in contrast to the patients with horizontal transmission for whom the levels of albumin (ALB) (P = 0.041), matrix metalloproteinase-3 (MMP-3) (P = 0.001) were high. The 47.2% of patients with MTCT and 36.8% of those with horizontal transmission had significant liver fibrosis (P = 0.013). Following antiviral therapy for 78 weeks, 21.2% and 38.0% patients with MTCT and horizontal transmission acquired hepatitis B e antigen (HBeAg) clearance, respectively (P = 0.043), and the virological response rates were 54.7% and 74.1% in the MTCT and horizontal groups, respectively (P = 0.005). MTCT was a risk factor for HBeAg clearance and virological response.@*CONCLUSION@#Adult patients with MTCT were more prone to severe liver diseases, and the therapeutic efficacy was relatively poor, which underlined the importance of earlier, long-term treatment and interrupting perinatal transmission.@*TRIAL REGISTRATION@#NCT01962155; https://clinicaltrials.gov.

Study on Clinical Consistency Evaluation in TCM Practice Guidelines for Prevention and Treatment of Biqiu (Allergic Rhinitis) / 中国中医药信息杂志

Zhangjiagang Hospital of Traditional Chinese Medicine, as a collaborative unit of the project, participated in the clinical consistency evaluation in project of TCM Practice guidelines for Prevention and Treatment of Biqiu (allergic rhinitis) organized by the State Administration of Traditional Chinese Medicine. The evaluation results showed that the guideline met the clinical practice requirements. This article summarized the clinical evaluation experience from the three aspects: The diagnosis in TCM and Western medicine is clear and definite, but the English translation needs to be considered; The definition of remission phase of Biqiu (allergic rhinitis) is difficult, to interfere by identifying the constitution is innovative; The advantages of "preventive treatment of diseases" in TCM should be fully played in role, improving the satisfaction of patients.

Anterior and posterior cruciate ligament reconstruction under arthroscopy:tendon autografts versus allografts / 中国组织工程研究

BACKGROUND: Most domestic studies have shown that there are similar curative effects of allogeneic and autogenous tendons in anterior or posterior cruciate ligament reconstruction. Both of them are good grafts that can achieve satisfactory outcomes. OBJECTIVE: To compare the clinical efficacies of allogeneic and autogenous tendons in anterior and posterior cruciate ligaments reconstruction under arthroscopy. METHODS: A total of 100 patients with knee cruciate ligament rupture undergoing anterior and posterior cruciate ligament reconstruction under arthroscopy were enrolled in this study, and randomly divided into two groups (n=50 per group): autogenous tendon group and allogeneic tendon group. The joint stability and mobility of the two groups were compared before operation and at discharge. The muscle strength recovery, Werner patellofemoral pain score and the Lysholm score of the two groups were evaluated and compared at 1, 3, 6, 9 months after discharge. RESULTS AND CONCLUSION: (1) Joint stability: The Lachman test positive rates and axial shift test positive rates in the two groups after operation were significantly better than the baseline (P < 0.05). At discharge, there were no significant differences in Lachman test positive rates and axial shift test positive rates between the two groups. (2) Joint range of motion: There were no significant differences in the joint ranges of extension and flexion between the two groups after operation. (3) Follow-up visit: Muscle strength, the Werner patellar pain scores and the Lysholm scores in the two groups were significantly improved at discharge (P < 0.05), but there were no significant difference between the two groups at 1, 3, 6, 9 months after discharge. In summary, autologous hamstring tendon and allogeneic tendon have the same clinical therapeutic effects in anterior and posterior cruciate ligament reconstruction under arthroscopy, both of which can achieve good short-term outcomes.

Analysis on relationship between influenza A (H1 N1) and meteorological conditions and its prediction model / 预防医学

Objective TostudytheinfluenceofmeteorologicalconditionschangingontheactivityintensityofinfluenzaA (H1 N1 )and to establish the prediction model of H1 N1 positive rate associated with meteorological factors.Methods TheinfluenzaA(H1N1)datafromAprilof2009toJanuaryof2011insentinelhospitalanddailymeteorologicaldatain the same period were collected,which were analyzed by Spearman correlation analysis.The prediction model was establishedusingChi-squaredautomaticinteractiondetector(CHAID).Results Weeklyaverageatmospheric pressure(r =0.50),highest atmospheric pressure (r =0.51 ),lowest atmospheric pressure (r =0.50),average temperature (r=-0.40),highest atmospheric temperature(r=-0.41),lowest atmospheric temperature(r=-0.39), precipitation(r=-0.23 )and average wind speed (r=-0.22 )were positively correlated with the activity intensity of H1N1(all P<0.05).Factors that affected H1N1 positive rate were lowest atmospheric pressure,average wind speed and precipitation(P<0.05 ).The prediction model of H1 N1 positive rate showed that the correct rate of prediction was 66.67%.Conclusion Lowestatmosphericpressure,averagewindspeedandprecipitationarecloselyassociatedwith the activity intensity of influenza A (H1 N1 ).CHAID method can be used to predict the H1 N1 epidemics.

The application of China Infectious Diseases Automated-alert and Response System in Zhejiang province, 2012 / 中华流行病学杂志

<p><b>OBJECTIVE</b>To analyze and evaluate the application of China Infectious Diseases Automated-alert and Response System(CIDARS)in Zhejiang province.</p><p><b>METHODS</b>Data through the monitoring program in 2012 was analyzed descriptively and compared with the incidence data in the same period as well information related to public health emergency events.</p><p><b>RESULTS</b>A total of 14 292 signals were generated on 28 kinds of infectious diseases in the system, in Zhejiang province. 100% of the signals had been responded and the median time to response was 0.81 hours. 123 signals (0.86%)were preliminarily verified as suspected outbreaks and 33 outbreaks were finally confirmed by further field investigation, with a positive ratio of 0.23% . Information related to regional distribution showed significant differences which reflecting a positive correlation between the numbers of diseases and the time of early-warning(r = 0. 97, P < 0.01). Distribution of information related to different types of diseases was also significantly different, showing a positive correlation between the prevalent strength of the disease and the amount of information in a specific area(r = 0.80, P < 0.01).</p><p><b>CONCLUSION</b>CIDARS had a good performance which could be used to assist the local public health institutions on early detection of possible outbreaks at the early stage. However, the effectiveness was different for different regions and diseases.</p>

Behavioral pattern in Chinese school-aged children with cleft lip and palate

OBJECTIVE@#To obtain descriptive information of behavioral pattern in Chinese school-aged children with cleft lip and palate.@*METHODS@#A total of 93 cleft lip and palate patients between the age of 6-11 year-old and treated at West China Stomatology Hospital were selected. And another 100 unaffected controls, matched for age and gender, were recruited randomly from a common primary school in Chengdu. Chart review of medical records was used to obtain psychosocial checklists. Scores were compared with published norms and controls to evaluate the risk of problems, separately for three diagnostic groups.@*RESULTS@#The patients group had lower scores of social and academic competencies, especially those with facial deformity or speech problem. No difference was found in the aspect of activity competency. All patients showed elevations in behavior problems. But the type of behavior problems varied in different genders.@*CONCLUSIONS@#Chinese school-aged children with cleft lip and palate are at raised risk for social and academic difficulties. Specific pattern of behavior problems displays differently depending on gender of the patient.

Evaluation on the performance of China Infectious Disease Automated-alert and Response System(CIDARS)in Zhejiang province / 中华流行病学杂志

Objective To evaluate the performance of China Infectious Disease Automatedalert and Response System(CIDARS). Methods A retrospective analysis was conducted on data related to the warning signals, the outcome of signal verification, the field investigation of CIDARS,and the emergent events reported through Public Health Emergency Events Surveillance System from July 1,2008 to June 30, 2010 in Zhejiang province. The performance of CIDARS was qualitatively evaluated by indicators on its sensitivity and rote of false alarm. Results In total, 26 446 signals were generated by the system which involving 17 diseases, with an average of 2.83 signals per country per week. Among all the signals, 99.95% of them were responded. 0.90% of the signals were judged as suspected events via the preliminary verification, and 30 outbreaks were finally confirmed by field investigation. The sensitivity of the system was 69.77% with the false alarm rate as 1.39%. Conclusion The system seemed to have worked on the outbreak early warning of infectious diseases and could directly reflect the anomaly event emerged from the infectious disease reporting system.However, more efforts should be paid to the following areas as how to decrease the false positive signals, select suitable thresholds and increase the quality of data in order to enhance the accuracy of the system.

Pulmonary fibrosis induces erectile dysfunction in rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the impact of pulmonary fibrosis on erectile function in rats and its mechanism.</p><p><b>METHODS</b>Forty 12-week-old healthy male SD rats were randomly divided into Groups A (4-week pulmonary fibrosis), B (6-week pulmonary fibrosis), C (4-week control, and D (6-week control). The models of pulmonary fibrosis were established by injection of bleomycin at 5 mg/kg in the trachea, while the controls were injected with normal saline only. At 4 and 6 weeks, all the rats were subjected to determination of the serum testosterone (T) level, arterial blood gas analysis, measurement of intracavernous pressure/mean arterial pressure (ICP/MAP), and examination of NOS activity and cGMP content. The mRNA expressions of eNOS, iNOS and nNOS in the corpus cavernosum penis were detected by real-time PCR, and that of eNOS analyzed by Western blot.</p><p><b>RESULTS</b>The 3 V and 5 V of the ICP/mapx100 in Group C were 16.37 +/- 2.19 and 27.19 +/- 3.18, significantly lower than 30.78 +/- 2.66 and 50.09 +/- 6.97 in Group A (P < 0.05); those in Group D were 10.17 +/- 1.31 and 17.40 +/- 1.74, significantly lower than 31.45 +/- 3.07 and 51.23 +/- 7.23 in Group B (P < 0.05), and so were they in Group D than in C (P < 0.05). PaO2 was significantly lower in Group C than in A ([75.50 +/- 13.87] mmHg vs [103.80 +/- 6.88] mmHg, P < 0.05) , and so was it in Group D than in B ( [83.60 +/- 5.50] mmHg vs [102.70 +/- 5.77] mmHg, P < 0.05). Group C showed a significantly increased serum T level as compared with A ([391.1 +/- 264.7] ng/dl vs [175.9 +/- 53.0] ng/dl, P < 0.05), so did Group D ([745.4 +/- 408.8] ng/dl) versus Group B ([177.8 +/- 52.3] ng/dl) and C (P < 0.05). NOS activity and cGMP content in the corpus cavernosum significantly decreased in Group C ([1.50 +/- 0.14] U/mg prot and [35.69 +/- 3.64] pmol/mg) compared with A ([2.66 +/- 0.39] U/mg prot and [51.10 +/- 7.22] pmol/mg) (P < 0.05), so did they in D ([1.40 +/- 0.20] U/mg prot and [34.55 +/- 4.30] pmol/mg) versus B ([2.75 +/- 0.36] U/mg prot and [52.15 +/- 6.86] pmol/mg) (P < 0.05), but neither showed any significant difference between Groups D and C (P > 0.05). The expression of the eNOS protein was significantly lower in Group C than in A (0.79 +/- 0.01 vs 0.87 +/- 0.01, P < 0.05), so was it in D than in B and C (0.71 +/- 0.02 vs 0.88 +/- 0.01 and 0.79 +/- 0.01, P < 0.05). The expression of eNOS mRNA was significantly higher in Group C than in A (4.46 +/- 0.92 vs 2.61 +/- 0.68, P < 0.05), but did not show any significant difference between D and B (2.79 +/- 0.60 vs 2.69 +/- 0.65, P > 0.05), nor did the expressions of nNOS mRNA and iNOS mRNA between the pulmonary fibrosis groups and the controls (P > 0.05).</p><p><b>CONCLUSION</b>Pulmonary fibrosis may induce erectile dysfunction by suppressing the expression of the eNOS protein and reducing NOS activity and cGMP content in the corpus cavernosum penis of rats.</p>

An analysis on the relationship between susceptibility to cirrhosis and polymorphisms at C-1350T and G-944C sites of CIITA gene promoter IV in chronic HBV carriers / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the relationship between the susceptibility to cirrhosis and the single nucleotide polymorphisms (SNPs) at C-1350T and G-944C loci of class II transactivator (CIITA) gene promoter IV in chronic HBV carriers.</p><p><b>METHODS</b>C-1350T and G-944C loci of CIITA gene promoter IV were analyzed by sequence-specific primer PCR (PCR-SSP) in 544 chronic HBV carriers and 125 non-HBV infected healthy blood donors.</p><p><b>RESULTS</b>Among the chronic viral hepatitis B patients, there were significantly decreased frequencies of CC and TG haplotypes, and significantly increased frequency of CG haplotype among patients with liver cirrhosis (CG vs. CC: chi2=8.274, df=1, P < 0.01; CG vs. TG: chi2 = 15.027, df =1, P <0.01). There were no significant differences in the frequencies of CC and TG haplotypes between chronic hepatitis B and liver cirrhosis patients (chi2 = 1.231, df =1, P < 0.05). There were significantly increased frequencies of CC/CC (Group 1) genotype and genotypes contained CG haplotype (Group 3), and significantly decreased frequencies chi2= 7.176, df = 1, P < 0.01; Group1 vs Group 4, chi2 = 19.818, df = 1, P < 0.01; Group 3 vs Group 2, chi2 = 11.423, df = 1, P < 0.01; Group 3 vs Group 4, chi2 = 34.226, df = 1, P < 0.01; Group 1 vs Group 3, chi2 = 0.009, df = 1; Group 2 vs Group 4, chi2 = 2.176, df = 1).</p><p><b>CONCLUSION</b>Polymorphisms at -1350 and -944 loci of CIITA gene promoter IV are associated with susceptibility to liver cirrhosis in chronic HBV carriers. The chronic HBV carriers bearing CC/CC genotype or genotypes containing CG haplotype progress into liver cirrhosis with more probability.</p>

Construction of eukaryotic expression vectors containing different haplotype cDNA of human CIITA gene / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To construct eukaryotic expression vectors containing three different haplotype cDNAs of human CIITA gene.</p><p><b>METHOD</b>cDNA fragments of three different CIITA haplotypes were obtained by inducing one or two single nucleotide mutations of wild type recombinant plasmid EBS-NPL-CIITA cDNA, which correspond to two non-homonymy single nucleotide polymorphism (SNP) sites in the coding region of human CIITA gene, using overlap extension PCR site-directed mutagenesis technology. The above-mentioned three haplotype cDNAs were respectively cloned to EBS-NPL-CIITA linearized vectors. Positive clones were identified by colonial PCR and restriction endonuclease digestion and were sent to be sequenced. Then eukaryotic expression vectors containing four different haplotypes and an empty vector EBS-NPL were transfected into HepG2 cells respectively. HLA-DR was detected by indirect cell immunofluorescence technique.</p><p><b>RESULTS</b>The cDNA fragments of three different human CIITA haplotypes were successfully constructed, and the eukaryotic expression vectors containing three different haplotype cDNAs of human CIITA gene were obtained. No expression of HLA-DR was observed in the original HepG2 cells and empty vector transfected HepG2 cells and the expression of HLA-DR emerged in the HepG2 cells transfected with four eukaryotic expression vectors.</p><p><b>CONCLUSION</b>The eukaryotic expression vectors containing three different haplotype cDNAs of human CIITA gene were successfully constructed, and they are essential for our further study of the functional differences of them.</p>

First record of Bursaphelenchus rainulfi on pine trees from eastern China and its phylogenetic relationship with intro-genus species / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Bursaphelenchus rainulfi isolated from dead pine trees in Zhejiang, China, is described and illustrated. It also provided some molecular characters of the Chinese population, including the PCR-RFLP and sequences of ITS region and D2-D3 expansion region of the large subunit (LSU) rRNA gene. Both the morphological characters and ITS-RFLP patterns match with the original description. The phylogenetic trees based on the 13 sequences of D2-D3 expansion region of the LSU rRNA gene and ITS region of Bursaphelenchus species were constructed, respectively, with the results showing the similar clades. The phylogenetic relationship based on the molecular data is similar to that with morphological characters. This is the first report of the species on pine wood in eastern China.

Study of the quasispecies dynamics of serum hepatitis B virus in a patient with acute exacerbations of chronic hepatitis B / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To investigate the quasispecies dynamics of hepatitis B virus (HBV) during the course of exacerbation and resolution of chronic hepatitis B in a patients.</p><p><b>METHODS</b>Five serum samples were collected from a patient with two episodes of exacerbation and resolution of chronic hepatitis B. A method of PCR-TA cloning-conformation sensitive gel electrophoresis (CSGE)-sequencing was performed to study the dynamic changes of HBV quasispecies in basal core promoter (BCP), precore and core regions of HBV genome.</p><p><b>RESULTS</b>Quasispecies complexity was 10 and 12 at the points of exacerbations, and 14 and 17 at the points of resolutions (t = 3.133, P < 0.05). Ratio of dominant quasispecies in HBV population was high (42.4% and 51.5%) during exacerbations and low (30.3% and 21.2%) during resolutions (t = 3.295, P < 0.05). All dominant quasispecies, except the one during the second resolution, carried core P5T, L60V, S155T, and precore G1896A mutations.</p><p><b>CONCLUSION</b>The composition of HBV quasispecies changes due to the change of host immune status, and immune pressure might lead to the selection of immune escape mutants.</p>

A multi-center clinical study of N-acetylcysteine on chronic hepatitis B / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To evaluate the effectiveness and safety of N-acetylcysteine (NAC) in treating chronic hepatitis B patients.</p><p><b>METHODS</b>144 patients with chronic hepatitis B (total bilirubin, TBil>170 mmol/L) from several centers were chosen for a randomized and double blind clinical trial. The patients were divided into a NAC group and a placebo group and all of them were treated with an injection containing the same standardized therapeutic drugs. A daily dose of 8 microgram NAC was added to the injection of the NAC group. The trial lasted 45 days. Hepatic function and other biochemistry parameters were checked at the experimental day 0 and days 15, 30, 45.</p><p><b>RESULTS</b>Each group consisted of 72 patients of similar demology and disease characteristics. During the trial, 28 cases of the 144 patients dropped out. In the NAC group, at day 0 and day 30, the TBil were 401.7 vs. 149.2 and 160.1+/-160.6. In the placebo group, the TBil on the corresponding days were 384.1+/-134.0 and 216.3+/-199.9. Its decrease in the NAC group was 62% and 42% in the placebo group. At day 0 and day 45 of treatment, the effective PTa increase rate was 72% in the NAC group and 54% in the placebo group. The total effective rate (TBil + PTa) was 90% in the NAC group and 69% in the placebo group. The parameters of the two groups showed a remarkable difference. The rate of side effects was 14% in the NAC and 5% in the placebo groups.</p><p><b>CONCLUSION</b>NAC can decrease the level of serum TBil, increase the PTa and reduce the time of hospitalization. NAC showed no serious adverse effects during the period of our treatment. We find that NCA is effective and secure in treating chronic hepatitis B patients.</p>

Betaine improves LA-PCR amplification / 生物工程学报

PCR is a powerful tool for the amplification of genetic sequences. It has been widely applied in molecular biology. It is generally used to amplify short segments (several hundreds basepairs to several kilobasepairs). It is difficult to amplify a long DNA segment. Based on the sequenced genes, it is known that most intact genes are very long. And intact gene is very important for the gene to express specially and effectively. Long PCR is a very useful tool to amplify intact genes for constructing special expression vectors. We have tried several chemicals to optimize long PCR system and found betaine was the best. Betaine, as an amino acid analogue with small tetraalkylammonium ions, could remarkably improve the amplification of long targets from the plant genome. The suitable concentration of betaine was between 1.0 mol/L and 2.5mol/L. We could effectively amplify a 9 kb DNA segment from maize genome DNA and a 16 kb DNA segment from plasmid. It was shown that different primers and different targets (different GC content) needed different concentrations of betaine. Betaine can reduce or eliminate non-special amplification. In the meantime we tried other additive chemicals, such as DMSO, glycerin, formamide. They were no notable results in long PCR.