The model of vaginal candidiasis in Kunming mice was constructed in order to search for the optima construction conditions and provide an economic animal model of Candida albicans (C. albicans) vaginitis. Estrogen benzoate (E2) was given to mice at different concentrations ranging from 0.0 to 0.05 mg/mouse (4 levels) beginning 72 h prior to vaginal inoculation, then mice were inoculated intravaginally with various concentrations of stationary-phase C. albicans blastoconidia (ATCC90028) (5 levels) in 20 microL of phosphate-buffered saline (PBS) in each E2 level. General state, scores of genital pathology, the hyphae and vaginal fungal burden (CFU) in vaginal lavage fluid, the hydrops rate of uterus and vaginal tissues for pathological section in mice were observed and obtained at day 2, 4, 7, 14 and 21 after inoculation. The results showed the infection rate in mice was related to the dosage of E2 and concentration of C. albicans blastoconidia. Additionally there was better cross-effect between the two treated factors. The infection rate was about 80% on the day 4, and could reach 100% on the day 7 until the end of experiment after inoculated intravaginally in groups of E2I3, E2 0.025 mg/mouse injected hypodermically and inoculated intravaginally with 5 x 10(4) C. albicans blastoconidia, and large amount of hyphae and blastoconidia could be observe in superficial layer tissue and canal of vaginal by PAS. From the results in our experiment it was concluded that E2I3 was the optima construction condition in kunming mice.

Objective To investigate the protective effects of the extract of ginkgo biloba(EGb)on the spiralganglion neuron(SGNs)in cochlea tissues on the hearing loss induced by noise in rats.Methods Thirty-six healthy animals were randomly divided into three groups:the normal control group(n=12).the noise exposured group(n =12)and the EGb treamment group(n=12).The control group received no noise and no medications.The other two groups were exposed to the noise of 110 dB SPL for consecutively 10 days,6 hours per day.The treatment group rats were injected with 10 ml/d EGb while the other two groups with 0.9%saline of the same amount.The experiment lasted for ten days.The rats were measured by auditory brainsterm response(ABR)before and after niose exposure.The ultrastructural changes of SGNs were detected by tranismision electron microscpoe(TEM) and the contents of malondiadehyde(MDA) and activities of superoxide dismutase(SOD)were also measured.Results Hearing were signifcantlly decreased in the experimental group.Nevertheless,EGb relatively reduced the contents of MDA while increased the activities of SOD.Conclusion EGb seems to be able to moderately pretect SGNs and to play a preventive and remedial role in noise-induced hearing loss.

Objective To evaluate and compare the results of different operative methods for different types of femoral neck fracture. Methods To assess the treatment effect of two methods. Some eases of femoral neck fracture of basal-neck type and via-neck type were treated with closed reduction internal fixation with two compression spiral screws(14 eases). Other eases(32 eases) of femoral neck fracture of sub-head type and head-neck type were treated with closed reduction internal fixation with two cannulated compression screws and bone graft flaped with quadrate muscle of thigh. Results 41 cases were followed up. The average healing time was 3 years and 10 months.The bone healing rate was 97.5% ,femoral head necrosis occurred in 2.4% ,excellent and good rate was 92.6%.Conclusion Cases of femoral neck fracture of sub-head type and head-neck type were treated with closed reduction internal fixation with two cannulated compression screws and bone graft flaped with quadrate muscle of thigh. This method has the advantage of reliable fixation, simple operation, less injury and little hemorrhage. It can improve the rate of fracture union and reduce the rate of avascular necrosis of femoral head. It is the optimal method for the treatment of femoral neck fractures.

Objective To study the effect of glycyrrhizin(GL) on the gene expression of high mobility group protein 1 (HMGB1) in hippocampus and serum.To evaluate the effect on the expression of neuron-specific nuclear-binding protein (Neu-N) in the hippocampus CA1,CA3 regions in the chronic stage of an immature rat epilepsy model.Methods Fifty-two 21 day-old SD rats were randomly divided into control group,model group Ⅰ and model group Ⅱ according to the random table method.Model group Ⅰ was induced epilepsy by kainic acid (KA),and the model group Ⅱ was pretreated with GL by intraperitoneal injection at 30 min before KA injection.According to the different observation time points,each group was divided into 4 subgroups:3 h,12 h,24 h and 7 d.Model group Ⅱ was divided into 3 subgroups:10 mg/kg,50 mg/kg,100 mg/kg,according to the different doses of GL.There were 3 animals in each subgroup.Score was performed according to the Racine score,and quantitative real-time polymerase chain reaction and Western blot were applied to detect the mRNA and protein expression of HMGB1 in the acute phase.Enzyme-linked immunosorbent assay(ELISA) was applied to measure the expression of HMGB1 in blood;immunohistochemical was applied to measure the expression of Neu-N in hippocampus in the chronic phase(7 d).Results Compared with model group Ⅰ,seizure onset time was obviously prolonged in model group Ⅱ [(24.08 ± 1.98) min vs.(33.39 ± 2.66) min],and the difference was statistically significant (t =9.231,P ＜0.05);Comparing KA model group Ⅰ with control group,the gene expression of HMGB1 significantly increased,and reached a peak at the time of 12 h (H =10.532,P ＜ 0.05),but the protein expression of HMGB1 was changed obviously and there was no significant difference (H =5.227,P ＞0.05).The expression of HMGB1 in the serum also significantly increased,especially at 12 h (H =6.897,P ＜0.05).At the time of 12 h after KA injection,the gene expression of HMGB1 in the hippocampus was significantly decreased in model group Ⅱ compared with model group Ⅰ (H =10.721,P ＜0.05) (especially in the 100 mg/kg model group).Also,the expression of HMGB1 in the scrum was obviously decreased (H =6.967,P ＜ 0.05) (especially in the 100 mg/kg model group).At the time of 7 d after KA injection,hippocampal neuron loss in model group.Ⅰ was significantly reduced compared with control group (P ＜ 0.05),and hippocampal neuron loss in model group Ⅱ was evidently decreased compared with model group Ⅰ (P ＜ 0.05),(especially in the 100 mg/kg model group in CA1,50 mg/kg model group in CA3).Conclusions In the immature rat temporal lobe epilepsy model,GL may have neuroprotective by inhibiting the synthesis and release of HMGB1,inhibiting inflammation further to restrain the loss of neurons in the chronic phase.

Linoleic acid (C18∶2n-6) and ?-linolenic acid (C18∶3n-3) are found widely in fungi, plants and some lower animals. However, they can not be synthesized in mammals due to lack of △12 and ?-3 fatty acid desaturases. To enable endogenous production of essential fatty acids in mammalian cells, here the stable expression of a Caenorhabditis elegans gene FAT-2 encoding △12 fatty acid desaturase in CHO cells was reported. First, the FAT-2 coding sequence was cloned by RT-PCR. To facilitate high level synthesis of heterogeneous protein, the codon usage of the fatty acid desaturase genes was optimized according to the codon preference of mouse by site-directed mutagenesis, 2 synonymous mutations were introduced into FAT-2 gene by overlapping PCR. The codon-modified gene was finally fused to pBudCE4.1 vector (Invitrogen) under the control of CMV promoter. The expression vector pBudCE-FAT2 was linearized with NheⅠ, and then transfected CHO cells, the cells were under Zeocin selection for nine days and then propagated, then the transfected cells were harvested. The genome and total RNA were isolated for PCR and Norhern blot ananlysis. The results revealed that FAT-2 gene has been integrated into the genome of CHO cells and expressed properly. Fatty acids of total cellular lipids were analyzed by gas chromatography. The results indicate that the expression and function of △-12 fatty acid desaturase resulted in accumulation of linoleic acid. The levels of linoleic acid in transgenic cells were 2.4-fold higher than those in wild-type cells. The moderate linoleic acid in CHO cells was derived from cell culture media uptaken by cell membrane. The results demonstrate that a heterogenous desaturase gene can function well in mammalian cells and prove that transgenic approach is an efficient strategy for changing fatty acid composition of mammals.

Objective To investigate the stimulating effect of exogenous hydrogen sulfide (H2S) on angiogenesis in glioblastoma (GBM). Methods Twenty adult Sprague-Dawley (SD) rats were randomly divided into two groups, glioma group (C6 glioma cell intracerebral implantation, n=10) and glioma-H2S group (C6 glioma cell intracerebral implantation and sodium hydrosulfide (NaHS) intraperitoneal injection, n=10). The tumor-bearing rat model was established by intracerebral injection of rat C6 glioma cells. After one week, normal saline was injected in glioma group and NaHS was injected in glio-ma-H2S group. Food and water were freely available during all phases of the experiment. After three weeks, rats were decapi-tated and brains were removed. HE staining was performed to show tumor structure and intratumoral angiogenesis. The immu-nohistochemical analysis was used to detect the expressions of CD34 and MMP-2, respectively. The microvessel density (MVD) in GBM was also measured. Results HE staining showed that the implanted tumors were predominantly spheroid with clear border and no capsule could be detected. The neovascular proliferations were observed in tumors. There were high-er expressions of CD34 and MMP-2 in glioma-H2S group. The value of MVD was significantly higher in glioma-H2S group than that of glioma group (P＜0.01). Conclusion Exogenous H2S serves as a stimulator of angiogenesis in the development of rat GBM, which may be related with the increased MMP-2 expression promoted by H2S.

Objective To establish an accurate method for estimating the indoor gamma dose rate from decorative stones.Methods Combining a room model with decorating conditions,the gamma dose rates and dose rate conversion factors (DCF) at 1 m above the floor in the room center were calculated with the Monte Carlo simulations,and the calculation results were verified through experiments.Based on the results,the limit of radionuclide contents in stone materials was further discussed.Results The DCF increases with the increase of area or thickness of decorative stones in the same room.The increase of DCF with the thickness of decorative stone is approximately linear.The DCF also increases with the area of decorative stones,but the increasing trend slows down.For the same decorative stones,the smaller the room,the larger the increase of gamma dose rate.Experimental results were consistent with the simulation results within ± 20％.Conclusions The increase of indoor gamma dose rate depends not only on the radionuclide contents,but also on the area and thickness of the decorative stones as well as the room size.The method used in this study can be used to estimate,more accurately than ever,the additional external exposure to residents due to decorative stones,and it provides a theoretical basis for revising the limit standard on radionuclide contents in decorative materials.

Objective To detect anti-HCV in serum of hepatic disease patients by performing the confirmatory test, and further to confirm HCV infection. Methods Two recombinant immunoblot assays (CWT and CHIRON RIBA HCV 3.0 Strip Immunoblot Assay) were used respectively to detect anti-HCV in 477 human serum samples, which comprised 350 HCV-infected patients' specimens, 7 none-A none-E hepatitis specimens, 30 HBV-infected patients' specimens, 30 hepatitis E virus infected patients'specimens, and 60 specimens drawn from blood donors. The latter three groups served as controls. Results A total of 120 control non-HCV-infected patients' specimens were negative when tested by both assays. Among 350 HCV-infected patients, 341 were positive and 9 were indeterminated by CWT assay; 343 were positive and 7 were indeterminated by CHIRON RIBA HCV 3. 0 SIA. Seven none-A none-E hepatitis specimens tested by both assays turned out to be 2 positive, 4 negative and 1 indeterminate. The consistency rate of these two assays was 99. 16% (Kappa=0.98). Conclusion CWT assay is highly coherent with CHIRON RIBA HCV 3.0 SIA assay in the methodology of anti-HCV antibody detection, which can be applied in the determination of HCV infection among none-A none-E hepatitis patients.

Objective To investigate the effect of the Ras-related tumor suppressor gene aplasia Ras homolog member Ⅰ (ARHI) on angiogenesis in hepatocellular carcinoma (HCC).Methods We generated stable cell lines overexpressing ARHI in Hep3B cells,which lack endogenous ARHI.Cell proliferation was assessed by the MTT assay.The effects of ARHI overexpression on tumor growth and angiogenesis were assessed.Because of the key role of mammalian target of rapamycin (mTOR)signaling in HCC progression,we also tested whether ARHI overexpression affected the mTOR pathway.Results Ectopic expression of ARHI significantly diminished cell proliferation in Hep3B cells (P＜0.01).ARHI overexpression significantly retarded Hep3B xenograft growth by 76.4 ％ in vivo,and caused a marked reduction in tumor angiogenesis assessed by CD31-stained microvessel count.Western blot analysis of the xenografts showed that ARHI overexpression substantially reduced the phosphorylation of two mTOR substrates,S6K1 and 4E-BP1,indicative of an inactivation of the mTOR pathway.Accompanying with the mTOR inactivation,the angiogenic factors,hypoxia-inducible factor 1 alpha and vascular endothelial growth factor,were significantly downregulated.Conclusion These data highlighted an important role for ARHI in controlling HCC growth and angiogenesis,therefore offering a possible therapeutic strategy against this malignancy.

Objective To evaluate the feasibility of MRI-guided vacuum-assisted breast biopsy in domestic clinical prac tice.Methods A total of 45 patients with 45 suspicious breast lesions were examined with MRI-guided vacuum assisted bi opsy.The complications were observed.And the followed-up was performed.Results In 45 suspected breast lesions,43 lesions (43/45,95.56％) were successfully sampled under MRI-guided vacuum-assisted biopsy and the pathologic diagnosis was obtained.For another 2 lesions (2/45,4.44％),the pathology diagnosis were not obtained.The time to perform MRI-guided biopsy was (45.87±8.41)min for one lesion.The 5 malignant lesions (5/45,11.11％) confirmed by biopsy pathology were consistent with postoperative pathological results.For the 38 benign lesions (38/45,84.44％) confirmed by biopsy pathology,there was no malignant performance found in MRI follow-up for 6 months.Because of successful MR imaging-guided core biopsy findings,the clinic treatments were changed in 14 patients (14/45,31.11％).Five cases (5/45,11.11％) with malignant tumors were diagnosed early.The surgical biopsy was avoided in 9 benign cases (9/45,20.00 ％) who suspected as malignant.In all 45 patients,no complications occurred,such as hematoma and infection.Con clusion MRI-Guided vacuum-assisted breast biopsy is sufficient and safe for obtaining histologic proof of lesions,and plays an important role in guiding clinic treatment of some lesions visible only by MRI.