Objective To establish a safe and effective method for obtaining fetus of rhesus monkey.Methods The pregnant process of rhesus monkey was monitored by B ultrasonography,and the pregnant rhesus monkeys in different stages of pregnant were randomly chosen.Mifepristone was subcutaneously injected to pregnant rhesus while misoprostol was put to their posterior fornix.The efficacy of this method was evaluated by the results of bishop score,the rate of pregnancy termination and the quality of total RNA extracted from brain samples of rhesus fetus.ResultsThe scores of bishop score and rates of fetus delivery of experiment group were both higher than that of control group(P

Objective To investigate the kinetic expression level of chemokines (MCP-1 and MIP-2) in vagina of a murine model of vulvovaginal candidiasis (VVC).Methods The estrogen-treated murine model of VVC was set up.Vaginal specimens were obtained in different duration after inoculation of C.albicans intravaginally.Semi-quantitative RT-PCR was applied to determine MCP-1 and MIP-2 mRNA levels in these tissues.Results Compared with the control mice treated with olive oil,persistent growth of C.albicans was found from the 2nd day to 21st day after inoculation in estrogen-treated mice.Significantly higher levels of MCP-1 mRNA were observed in vaginal tissues in infected estrogen-treated mice than those in other 2 groups,infected but non estrogen-treated mice and estrogen-treated but uninfected mice.The high level of MCP-1 mRNA maintained from the 4th day to 21st day in infected estrogen-treated mice.It was also found that levels of MIP-2 mRNA were significantly higher in the vagina in the 2nd day in 3 groups of mice than those in naive mice,however,no significant difference was shown among 3 groups throughout the study period.Conclusion High level of MCP-1,rather than MIP-2,may be associated with susceptibility to VVC.

BACKGROUND: Frenulum of prepuce of penis contained many nerve terminals is an extremely sensitive region. If the frenulum is injured in circumcision or other operations, the complication, such as postoperative spontaneous pain of penis, sexual disturbance and so on, will occur. But there still is no define explanation for this up to now.OBJECTIVE: To observe the distribution of immunoreactive nerve terminal of calcitonin gene-related peptide (CGRP) in prepuce of penis and frenulum of prepuce of adult SD rats, and look for the source of CGRP immunoreactive nerve terminal in frenulum of prepuce.DESIGN: A single sample trial.SETTING:Department of Anatomy, School of Medicine, Zhejiang University.MATERIALS: The experiment was performed at Department of Anatomy,School of Medicine. Zhejiang University from September 2004 to May 2005. A total of 20 adult male SD rats were selected, and were raised in warm, quiet, photophygous environment for 1 week before the trial so as to make the rats fit for the environment and maintain their basal state.METHODS: The rats were assigned randomly into 2 groups. Ten rats in the first group were treated with the immunohistochemical method to observe the distribution of CGRP immunoreactive nerve terminal in prepuce of penis and frenulum of prepuce of adult rats. Ten rats in the second group were treated with fluorogold (FG) retrograde labeled combined with CGRP immunofluorescence labeled method to look for the source of CGRP immunoreactive nerve terminal in frenulum of prepuce of penis. MAIN OUTCOME MEASURES: ①The morphology and distribution of CGRP immunoreactive nerve terminal in prepuee of penis and frenulum of prepuce of adult SD rats were observed under light microscope. ②The distributive density and difference of CGRP immunoreactive nerve terminal in prepuce of penis and frenulum of prepuce were detected and compared (represented by A). ③Morphology and distribution of FG retrograde labeled -positive, CGRP single-labeled positive and FG/CGRP double-labeled positive neurons in dorsal root ganglion were observe under fluorescence microscope. ④Mean quantity of FG retrograde labeled positive, CGRP single abeled positive and FG/CGRP double-labeled positive neurons in dorsal root ganglion was counted.RESULTS: Totally 20 rats were involved in the analysis of results. ① Amber-coloured CGRP immunoreactive nerve terminal appeared in prepuce of penis and frenulum of prepuce of adult rats. These nerve terminal mainly occurred in basal layer of epidermis and papillary layer of dermis, distributed as twig shape or intestiniform; mostly of them were bundled, different in length, and some of them showed enlarged nodosity. ②The distributive density of CGRP immunoreactive nerve terminal in frenulum of prepuce of penis was significantly larger than that in prepuce of penis (2.15±0.32, 1.02±0.22,t =-2.03,P＜0.01). ③Combined with the FG retrograde labeled method it was found that these nerve terminal was derived from neurons of dorsal root ganglion opposed to the sixth lumbar spinal cord and the neurons of dorsal root ganglion opposed to the first acral spinal cord. FG retrograde labeled positive neurons differed in length. The cell body showed round or orbicular-ovate, without obvious prominence. Bright inaurate fine particle appeared in cytoplasm, no label in nucleus. Most cells arranged in line along nerve tract or diffusedly distributed. Most CGRP single-labeled positive neurons were middle or small cells found by CGRP immunofluorescence labeling. Dyeing was too dark.Reaction product distributed evenly in cytoplasm, which showed bright dark green (FITC labeled color). The same positive section was observed comparatively under different excitation light. It was found that FG/CGRP double-labeled positive cells were middle or small, and its amount accounted for a half of the total number of FG retrograde positive cells.CONCLUSION: CGRP may participate the transmission of sensory information in prepuce of penis and frenulum of prepuce of rats. The CGRP immunoreactive nerve terminal in frenulum of prepuce of penis of rats is sourced from neurons of dorsal root ganglion opposed to the sixth lumbar spinal cord and the neurons of dorsal root ganglion opposed to the first sacral spinal cord.

Objective: To investigate the effect of human chorionic gonadotrophin (hCG) on the gene expression of migration inhibitory factor (MIF) in human peripheral blood mononuclear cell (PBMC). Methods: The healthy human PBMC was cultured with hCG at 37 ℃, 5%CO_2 for 2 hours. The mRNA of harvested cells was isolated. The MIF mRNA was detected by real-time RT-PCR. Results: In a certain range of doses, the mRNA expression of MIF significantly increased following the increase of hCG in a dose depandent manner, and it reached to a peak 1-2 hours after culture, then returned to the minimum level after 8 hours. Conclusion: In a certain range of doses, hCG can increase the mRNA expression of MIF. This effect is correlated with reacting time. It is suggested that hCG may involve in immune response by up-regulating the production of cytokines by PBMC.

Objective Lung cancer has been the most common cause of cancer death worldwide.More than three quarters is diagnosed at advanced stage.Nearly one half has distant metastases outside the chest cavity.It has been recognized that oligometastatic state exists in natural history of malignant tumors,which has significantly better prognosis than those with polymetastases.The eighth edition of TNM staging system defined NSCLC with a single metastasis as a new separated stage(M1 b).Long-term survival of oligometastatic NSCLC patients treated with local control methods has been reported in literatures.But whether surgical resection of liver metastasis has benefit or not remains unknown.We performed a systematic review of surgical therapy for NSCLC with limited liver metastases.

Objective To study port-externalized tissue expansion technique and to provide a safe,simple and effective skin tissue expansion for extensive skin lesion reconstruction.Methods A lot of 247 patients with extensive skin lesion were chosen for the study.400-1000 ml tissue expanders were used for overexpansion and repeated expansion.Expanders were inserted into subcutaneous pockets through mini-invasive incisions and intermittent injection was carried out 4 days after first stage operation.After expansion finished,expanded skin flaps were transplanted for skin lesion reconstruction.During inflation period,complications and expander failures were observed.Results In 741 expanders of 247 patients,complication rate was 16.5 ％ with total 122 cases of complications.Expansion success rate was 98.9 ％ with 8 expander failures.Expanders were inflated successfully between 64-113 days,with average 92.7 days.The inflated saline volume was 2.38 times of marked expander capacity.In this group of 733 expanded skin flaps,721 flaps were well survived with 98.4％ success rate.276 expanded flaps of 116 patients were followed up for one to five years post-operation.The width of incision scar was between 0.3-1.2 mm.Conclusions The port-externalized overexpansion with giant expanders is a safe,simple and feasible technique,which is worthy of further clinical application.

Objective:To establish the quality standard for Chidan Ganxian granules. Methods:TLC was used to identify Paeoni-ae Radix Rubra, Rhizoma Polygoni Cuspidati, Radix Salviae Miltiorrhizae, Herba Saururi and Radix et Rhizoma Rhei in Chidan Ganx-ian granules, and the content of paeoniflorin was determined by HPLC. The stationary phase was an Apollo C18 column ( 150 mm × 4. 6 mm, 5 μm), the mobile phase was acetonitrile-water(12. 5∶87. 5), the flow rate was 1. 0 ml·min-1, the detection wavelength was at 230 nm, and the temperature was 30℃. Results:The characteristic spots of the granules were the same as those of the standard samples without any interference from the negative control. Paeoniflorin had a good linear relationship within the range of 0. 120-1. 436 μg(r=0. 999 4), and the average recovery was 99. 8% with RSD of 1. 80%(n=6). Conclusion:The method is simple, ac-curate, reliable and reproducible, which can be used in the quality control of Chidan Ganxian granules.

Objective To analyze the types and risk factors of community-acquired infections (CAI)in diabetic patients by system analysis method of evidence-based medicine.Methods China National Knowledge Infrastructure (CNKI),Wanfang database,VIP database were searched by computer,domestic published researches on CAI and related risk factors in dia-betic patients were aggregated,Meta-analysis was conducted by stata 1 1 .0 software.Results A total of 1 2 literatures were included in the study .The average rate of CAI in diabetic patients was 39.55% (22.12%-55.86%).The major infec-tions were respiratory system infection(40.74%),urinary tract infection(27.35%),tuberculosis(10.80%),skin and soft tissue infection(9.19%),and hepatobiliary system infection (5.57%).Stratified analysis on risk factors revealed that OR and OR95%CI of chronic complication,age,disease course,glycemic control,gender,type of diabetes,subtype of ketoac-idosis was 1.63(1.45,1.82),1.30(1.19,1.42),1.47(1.35,1.61),0.68(0.61,0.76),0.69(0.64,0.75),1.37 (1.13,1.66 )and 0.87(0.62,1.23),respectively.There was no publication bias and combined results were stable. Conclusion The main CAI in diabetic patients are respiratory system infection,urinary tract infection,tuberculo-sis,skin and soft tissue infection,and so on ;several factors,such as female,older age,long-term disease course, poor glycemic control,and complication,can contribute to the increase of CAI in diabetic patients.

Objective To construct the DNA vaccine pIRES2-MLAA34-HSP70,and to detect its immune effect.Methods The acute monocytic leukemia associated antigen gene MLAA-34 and heat-shock protein (HSP)70 gene were extracted by using RT-PCR.The specific overlapping primer was designed,and the fusion gene MLAA34-HSP70 was amplified by using SOE-PCR technique.Then the DNA vaccine pIRES2-MLAA34-HSP70 was constructed,and BALB/c mice were immunized with this DNA vaccine.The splenic lymphocyte killing activity was detected by using MTT,levels of IL-2,IL-4 and IFN-γ were also detected by using ELISA.Results The MLAA34-HSP70 gene (2 956 bp) and the DNA vaccine pIRES2-MLAA34-HSP70 was amplified and constructed successfully.The killing efficiency of DNA vaccine pIRES2-MLAA34-HSP70 in U937 cells was significantly higher than that in other experimental groups and control group (P＜0.01),and levels of IL-4,IL-2 and IFN-γin DNA vaccine pIRES2-MLAA34-HSP70 group were significantly higher than those in the other experimental groups and control group (P＜0.01).Conclnsion The DNA vaccine pIRES2-MLAA34-HSP70 is constructed successfully.It is shown that the DNA vaccine induces strong humoral immunity,which could enhance immune responses to tumor cells and specificlly kill MLAA34 positive cells.

Hemagglutinin (HA) contains a head domain with a high degree of variability and a relatively conserved stem region. HA is the major viral antigen on the surface of the influenza virus. To define the biologic activities of chimeric HA bearing different head domains and stem regions or their potential use, a HA chimeric gene containing the head domain of the H7 subtype virus and stem region of the H3 subtype virus was modified and expressed using a baculovirus expression vector. Then, the secreted protein was purified and its biologic activities characterized. Approximately 1.4 mg/mL cH7/3 HA could be obtained, and its molecular weight was ≈ 70 kD. The trimer form of cH7/3 protein had hemagglutination activity and could be recognized by specific antibodies. The method described here can be used for further studies on the screening of HA stem-reactive antibodies or the development of vaccines with conserved epitopes.
Antibodies, Viral ; immunology ; Baculoviridae ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Hemagglutination ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; immunology ; Humans ; Influenza Vaccines ; genetics ; immunology ; Influenza, Human ; prevention & control ; virology