Objective:To express human interferon-?2b gene and to explore the feasibility of expressing human gene in plant cells.Methods:The hIFN-?2b coding sequence was amplified by PCR with specific primers and plasmid pBV889 was used as a template,subcloned into middle vector pMD18-T and binary vector pBI121 to obtain plant expression vector pBIFN. The pBIFN was transformed into Agrobacterium tumefaciens strain LBA4404. Then hIFN-?2b gene was introduced into Ginseng callus cells via Agrobacterium-mediated transformation. The positive cells were screened by G418. The transgenic Ginseng calli were confirmed by PCR,RT-PCR,Western blot and WISH/VSV system.Results:Stable integration of the hIFN-?2b gene in the Ginseng callus cells′ genome was confirmed by PCR analysis. RT-PCR analysis showed that there were transcription products. Western blot implied that the given protein was hIFN-?2b. WISH/VSV system assay showed that the expressed hIFN-?2b possessed relatively lower bioactivity.Conclusion:HIFN-?2b has been expressed in transgenic Ginseng calli, which facilitates further investigation of improving the curative effect of orally administered hIFN-?2b.

Objective To observe the clinical efficacy and safety of low-dose tacrolimus (TAC) combined with tripterygium(TW) in treatment of steroid-resistant nephrotic syndrome (SRNS). Methods The patients, who were diagnosed with mesangial proliferative glomerulonephritis (MesPGN) and focal segmental glomerulosclerosis (FSGS) by biopsy and failed to respond to a 3 month treatment with prednisone (1 mg/[kg • d], maximum 60 mg/d, were randomly divided into 2 groups (TAC+TW group and TW group). Initially TAC+TW group took TAC 0. 05 mg/(kg • d) 2 h after meal at a 12 h interval; the plasma TAC level was examined after 3 days and was kept at 1. 5-4 ng/ml; meanwhile, TW was given at 60 mg/d before meal. TW group only took TW(60 mg/d before meal). The efficacy, adverse reactions and plasma TAC levels were observed in each group. Results (1) Totally 20 patients met the recruitment criteria, including 11 in the TAC+TW group and 9 in the TW group. The age, sex, time of onset, blood pressure, 24 h urine protein, serum albumin, creatinine, cholesterol, triglyceride, fasting blood glucose, renal pathology types and period of prednisone use were similar between the two groups. (2) In TAC+ TW group the urine protein began to decrease one month later; 12 months later,8 cases had complete remission (72. 7%), 2 had partial remission (18. 2%),and 1 was ineffective (9. 1%), with a total effective rate of 90. 9%. I n TW group the urine protein also began to decrease one month later; 12 months later only 2 cases had complete remission (22. 2%), 4 had partial remission (44. 5%), and 3 were ineffective (33. 3%), with a total effective rate of 66. 7%. (3)In TAC + TW group the plasma protein was significantly higher than that before treatment; the plasma protein recovered to normal level after 6 month treatment. However, there was no significant increase in TW group. The pre- and post-treatment serum creatinine levels were similar between the two groups. (4) The incidence rates of adverse reactions were not significantly different between the two groups. Conclusion Low dose TAC combined with TW can effectively decrease proteinuria in patients with SRNS, generating a higher rate of clinical remission; meanwhile, the patients have a good tolerance and less adverse reactions.

Tegument protein VP22 is encoded by Pseudorabies Virus (PRV) UL49. To identify the nuclear localization signals of UL49, it is necessary to determine the transport mechanism and biological functions of the VP22 protein. In this study, we identified two nuclear localization signals from UL49, NLS1 (5RKTRVA ADETASGARRR21) and NLS2 (241PGRKGKV247). The functional nuclear localization signal (NLS) of UL49 was identified by constructing truncated or site-specific UL49 mutants. The deletion of both NLS1 and NLS2 abrogated UL49 nuclear accumulation, whereas the deletion of NLS1 or NLS2 did not. Therefore, both NLS1 and NLS2 are critical for the nuclear localization of UL49. And our resuts showed that NLS2 is more important in this regard.
Animals ; COS Cells ; Cell Nucleus ; metabolism ; virology ; Cercopithecus aethiops ; Herpesvirus 1, Suid ; chemistry ; genetics ; metabolism ; Humans ; Nuclear Localization Signals ; Protein Transport ; Pseudorabies ; metabolism ; virology ; Viral Structural Proteins ; chemistry ; genetics ; metabolism

Objective To investigate pathogene city, replication and expression of hepatitis G virus (HGV) in rhesus monkey infected with HGV RNA genome or HGV RNA positive sera. Methods A full length cDNA clone of HGV was constructed. Rhesus monkey BY1 was inoculated intrahepatically with genomic RNA from this HGV clone resulted in viral replication. HGV RNA positive sera from BY1 were intravenously inoculated into rhesus monkeys BM1, and sera from BM1 were intravenously inoculated into BB1 in series. Sera were collected weekly or bi weekly and liver biopsies were performed regularly. RT PCR, in situ hybridization and immunological, serological, histological assays were carried out to study the infectivity and pathogenecity of HGV. Results The serological and pathological results indicated that all of the 3 rhesus monkeys developed HGV viremia and had slightly elevated alanine transaminase levels (up to 418 IU/ L) during the period of experiment. HGV RNA became positive at the 3 rd , 8 th and 3 rd week post inoculation in the animals BY1, BM1 and BB1 respectively, and existed up to 21 weeks. The histology, immunohistochemnistry, and in situ hybridization in the liver tissues of the inoculated animals also showed that there was a mild hepatitis with HGV E2 expression in cytoplasm of hepatocytes. RT PCR and quantitative PCR showed that HGV could replicate in liver.Conclusions The genomic RNA from HGV full length cDNA is infective to the rhesus monkeys resulting in mild hepatitis. Infection and the transmission of the HGV in the rhesus monkey provide an appropriate animal model for the study of HGV.

Academic and scientific research are important indices which refer to the medical postgraduates' training quality.But research methods and statistical knowledge are important influencing factor for them. Understanding the demands of statistical knowledge for medical postgraduates can provide reference for adjusting the setting of teaching contents and satisfy the needs of the students.The results have showed that two kinds of medical postgraduates' knowledge of statistical knowledge is weak,while postgraduates with different types have different requirements for statistical knowledge.Therefore more attention should be paid to the needs of the learners of postgraduates with different types to provide more pertinent and effective teaching support service.

Objective　To analyze the influence of multiple variable factors on the occurance rate of pneumothorax associated with transthoracic needle aspiration biopsy of the lung. Methods　Fluoroscopically guided lung biopsies were performed in 46 patients. Variable factors were analyzed including lesion size, location, number of puncture, presence of emphysema and patients position after needle biopsy of the lung. Results　Pneumothorax occurred at 9 (19.6%) of 46 patients and that occurred at 4 (44.4%) of 9 emphysematous patients. Among them 2 necessitated chest drainage tube placement. The pneumothorax occurance rate was 30% (3/10) for lesions of diameter 3cm or less in size. In the dependent group, pneumothorax occurred in 4 of 20 patients (20%). In the nondependent group, pneumothorax occurred in 5 of 26 patients (19.2%). Conclusion　The correlation showed that increasing frequency of pneumothorax with decreasing size of lesions. An increased rate of pneumothorax was correlated with presence of emphysema. Patients with emphysema are more likely to have a symptomatic pneumothorax. No significant differences were found in the incidence of pneumothorax between patients placed with the puncture site dependent after biopsy and those placed with the puncture site nondependent.

BACKGROUND:Proto-oncogene c-Src plays an important role in regulating cardiovascular diseases such as hypertension. At present, there were no studies concerning exercise intervention effects on c-Src expression in aortic endothelial cels so as to regulate hypertension. OBJECTIVE: To observe the effects of aerobic exercise on c-Src mRNA expression and c-Src activity in the aorta blood vessel endothelial cels of spontaneous hypertensive rats. METHODS: A total of 8 male Wistar rats were considered as normal control group. Sixteen spontaneous hypertensive rats were randomly assigned to 8 rats as spontaneous hypertension group and 8 rats as spontaneous hypertension exercise group. Rats in the spontaneous hypertension exercise group carried on 90 minutes unloaded aerobic swimming every day, 6 days a week, for 8 weeks. The rats in the normal control group and spontaneous hypertension group did not swim. Blood pressure of rats was measured once a week. 8 weeks later, the c-Src mRNA expression and c-Src activity were determined in aortic vascular endothelial cels of rats in each group. RESULTS AND CONCLUSION: Compared with spontaneous hypertension group, blood pressure was lower, but c-Src mRNA expression and c-Src activity were significantly higher in the spontaneous hypertension exercise group. The c-Src activity and c-Src mRNA expression were higher in the spontaneous hypertension exercise group than normal control group and spontaneous hypertension group (P < 0.01). Results indicated that aerobic exercise can promote the increase in c-Src activity and c-Src mRNA expression in aortic endothelial cels of spontaneous hypertensive rats.

As an important ABC transporter, breast cancer re-sistance protein ( BCRP) plays an important role in tumor multi-drug resistance. Many laboratories are focusing on BCRP to re-verse multidrug resistance. We summarize in the paper the re-search progress on the regulation of BCRP expression, subcellu-lar localization, ATP-dependence, inhibition or modulation of its transport activity and potential clinical treatment strategies in or-der to provide theoretical support and some new research ideas for the reverse of multidrug resistance in clinic.

Budd-Chiari syndrome (BCS)is a rare liver disease,and there are mutual influences between BCS and pregnancy.The rela-tionship between BCS and pregnancy is systematically discussed,and the points we should pay attention to in the diagnosis and treatment of this disease are pointed out as follows:First,pregnancy is a risk factor for BCS;pregnant women with the clinical manifestations of portal hy-pertension and (or)inferior vena cava hypertension should be highly suspected of having BCS.Second,pregnancy is risky in BCS patients;pregnancy is allowable in BCS patients who have a stable condition after treatment,but they should be kept under rigorous surveillance dur-ing pregnancy and after delivery to avoid thrombosis recurrence.Third,BCS may lead to infertility in women;patients with unexplained in-fertility should be evaluated by abdominal ultrasound and computed tomography to clarify the diagnosis of BCS.

CD40 Ligand ; pharmacology ; HL-60 Cells ; drug effects ; Humans ; fas Receptor ; metabolism