Objective To investigate the optimal condition for in vitro cultivation of Trichomonas vaginalis for obtaining a better harvest of T\^vaginalis. Methods An isolate of T\^vaginalis from clinical specimens was cultivated in three different media with initial inoculation of 9\^0?10\+4/ml under pH 5\^6. Results There was distinct difference after 96h incubation in the cumulative harvest of T.vaginalis. The highest harvest was received in cysteine/liver/peptone/maltose medium, followed by the liver/peptone/maltose medium and soybean/liver/peptone/maltose medium. Conclusion The cysteine/liver/peptone/maltose medium may be a suitable environment for in vitro multiplication of T.vaginalis.

Objective To investigate clinical presentations,laboratory examinations,magnetic resonance imaging (MRI) appearances and treatment of hypertrophic cranial pachymeningitis (HCP).Methods The clinical data of 13 patients with HCP receiving comprehensive therapy in Huashan Hospital from January 2007 to January 2013 were analyzed retrospectively.Results The onset of HCP was mostly chronic with an average duration of 26.7 months.The main clinical manifestations of the 13 patients were chronic headaches (12/13) and cranial nerve paralysis (12/13).Inflammation markers and cerebro-spinal fluid (CSF) protein levels increased in patients with HCP and gradually became normal after the treatment.The MRI demonstrated local or diffused thickened dura located in tentorium (10/13),falx cerebrum (5/13),frontal lobe (4/13),temporal lobe (7/13) and parietal lobe (4/13).The signal intensity was isointense on T1-weighted MR images and hypointense on T2-weighted MR images.Enhanced MR images showed conspicuous enhancement of the dural edges.Corticosteroid therapy improved the clinical symptoms in 12 of 13 patients.Conclusions HCP typically causes headache and paralysis of multiple cranial nerves.Enhanced MRI shows characteristic manifestations.At present corticosteroid therapy is the treatment of choice followed by immunosuppressive agent and radiotherapy.

OBJECTIVE: To explore the curative effect of combination therapy with salvianolate injection (SAFI) and aspirin on anti-platelet aggregation and blood coagulation function, provide the experimental evidence for clinical consequence use of combination therapy with traditional Chinese medicine and western medicine. METHODS: The bleeding time was measured by cutting tail evaluating of combination of drugs; platelet maximum aggregation rate induced by ADP, AA, collagen and thrombin in rats was determined by absorbance method using a microplate reader; blood coagulation of the four indicators were measured by the semiautomatic blood agglutination instrument. RESULTS: SAFI had nearly no effects on aspirin group's bleeding time. The effect on blood platelets aggregation compared with control group, SAFI group and aspirin group could significantly reduce platelet aggregation induced by ADP, AA, collagen and thrombin in normal rats(P<0.01). Combinative group could significantly increase aspirin group's anti-platelet aggregation induced by ADP, AA, collagen and thrombin in normal rats(P<0.01, P<0.05). Coagulation function SAFI can reduced aspirin group's TT after the combination therapy. CONCLUSION: SAFI can increase anticoagulation effect of aspirin, and have no effect on aspirin's bleeding risk.

Adult ; Dioxins ; Glutathione Peroxidase ; blood ; Humans ; Lipid Peroxidation ; drug effects ; Male ; Malondialdehyde ; blood ; Middle Aged ; Occupational Exposure ; Oxidative Stress ; drug effects ; Superoxide Dismutase ; blood

OBJECTIVETo gain an insight into the large intragenic hMSH2 and hMLH1 deletions in Chinese hereditary nonpolyposis colorectal cancer (HNPCC) families.

METHODThe large intragenic hMSH2 and hMLH1 deletions in 17 probands of HNPCC families were detected with multiplex ligation-dependent probe Three large intragenic hMSH2 deletions of examplification (MLPA) and GeneMapper techniques.

RESULTSon 8, exon 1-6, and exon 1-7 were found in three families respectively, and no hMLH1 deletion was found. The deletions accounted for 19% of the total hMSH2 and hMLHI germline pathogenic mutations.

CONCLUSIONSThe incidence of large intragenic mismatch repair (MMR) genes deletions is relatively higher in Chinese families, and hMSH2 deletions may be more common. It is necessary to detect the large intragenic MMR genes deletions in the molecular detection of HNPCC.

Adaptor Proteins, Signal Transducing ; genetics ; Colorectal Neoplasms, Hereditary Nonpolyposis ; genetics ; Female ; Gene Deletion ; Humans ; Male ; MutL Protein Homolog 1 ; MutS Homolog 2 Protein ; genetics ; Nuclear Proteins ; genetics ; Nucleic Acid Amplification Techniques ; methods ; Pedigree

Objective:To establish a real-time fluorescent quantitative PCR for the detection of torque teno virus types 7 (TTV7), 8 (TTV8) and 10 (TTV10) and analyze its performance in clinical sample detection.Methods:Specific primers were designed based on the gene sequences of TTV7, TTV8 and TTV10 in GenBank. Recombinant plasmids of pMD19-T-TTV7, pMD19-T-TTV8 and pMD19-T-TTV10 were constructed and used as positive standard control to establish a real-time fluorescent quantitative PCR based on FAM-Eclipse probe method. The specificity and sensitivity of the established method were evaluated. Moreover, it was validated in terms of clinical sample detection.Results:The standard curve equations of the real-time fluorescent quantitative PCR for detecting TTV7, TTV8 and TTV10 were y=-0.340 2 x+ 114.780 0 ( R2=0.998 8), y=-0.351 1 x+ 114.940 0 ( R2=0.995 3) and y=-0.348 9 x+ 115.020 0 ( R2=0.991 7), respectively, and there was no cross-reaction with other viruses. The detection sensitivity of the established method for TTV7, TTV8 and TTV10 were 108 copies/μl, 84 copies/μl and 98 copies/μl, and the positive detection rates in clinical pediatric serum samples were 10.9%, 2.1% and 4.3%, respectively. Conclusions:The established real-time fluorescent quantitative PCR for detection of TTV7, TTV8 and TTV10 was featured by strong specificity and high sensitivity, which could be used for rapid TTV detection in clinical serum samples.

Objective To investigate the effect of silenced RBM8A gene on the biological behavior (proliferation, migration, and apoptosis) of human endometrial cancer HEC-1A cells and its possible mechanism. Methods The hairpin shRNA targeted by the RBM8A gene was designed, and the best shRNA silencing fragment was screened. The recombinant lentiviral interference vector carrying the target gene was constructed and used to infect HEC-1A cells. Cells with stable knockdown of RBM8A gene were screened by puromycin as the experimental group (shRBM8A), while the shRNA of nonsense sequence was designed as the control group (shControl). CCK-8 method was used to detect cell proliferation, and flow cytometry was used to detect cell apoptosis. Transwell assay was used to detect cell migration and invasion. Western blot was used to analyze the expression of apoptosis-related proteins and EMT signal transduction pathway related proteins. Results In comparison with the shControl group, after RBM8A knockdown, HEC-1A cell proliferation was reduced, apoptosis was increased, migration and invasion ability were significantly inhibited (P < 0.05), the expression of apoptosis-related proteins cleaved caspase 9 and caspase 3 increased, EMT-related protein E-cadherin expression increased, and Vimentin expression decreased. Conclusion RBM8A gene silencing can inhibit the proliferation, migration, and invasion and promote the apoptosis of endometrial cancer cells. The inhibition of EMT signal transduction pathway may be its mechanism.

Measurement of platelet-associated imunoglobulin (PAIg) has frequently been applied for the diagnosis of idiopathic thrombocytopenic purpura (ITP) and other immune thrombocytopenias. In the present study, a flow cytometry (FCM) analysis has been used to detect and characterize PAIg in 47 patients with ITP and Evans' syndrome, 13 patients with non-immune thrombocytopenia, 10 patients with autoimmune hemolytic anemia (AIHA) whose platelet counts were in normal range, and 31 healthy volunteers. With FCM measurement, mean fluorescence intensity (MFI) of platelets from patients with ITP and Evans' syndrome (2.26 +/- 2.29) was significantly higher than those from non-immune thrombocytopenia (0.33 +/- 0.39), AIHA (0.17 +/- 0.07) and control subjects (0.25 +/- 0.15) (P < 0.01). Meanwhile, the percentage of positive platelets of patients with ITP and Evans' syndrome [(44.1 +/- 29.0)%] was also higher than those of non-immune thrombocytopenia [(17.5 +/- 9.4)%], AIHA [(10.7 +/- 7.5)%] and control subjects [(16.6 +/- 8.4)%] (P < 0.01). In addition, some peak shape abnormality appeared (double peaks and peak tail) in the histogram of fluorescence intensity (log) of 11 patients (23.4%) with ITP and Evans' syndrome either alone or accompanied with quantitative alteration of MFI and/or positive platelet percentage. In seven cases, the peak shape abnormality was the unique characteristic that could be detected and have never been seen in normal platelets. This phenotypic alteration perhaps reflects the existence of different platelet populations and could be of diagnostic value. Totally, the positive result of FCM measurement in patients with ITP and Evans' syndrome was 87.2%, slightly higher than 83.0% positive rate with ELISA method, without statistical difference. The correspondent rate of the results of these two analytical settings was 85.1%. This study shows that FCM assay is a rapid and sensitive method for the measurement of PAIg and seems to be suitable as a novel routine diagnostic technique of immune thrombocytopenia.

Desmoglein,which is a kind of Ca2+ dependent desmosomal cadherins protein,is a major part of the desmosomes.The desmoglein that distributes in the epithelium,myocardium and other tissues plays a very important role in the cell junction.In recent years,the detection of the abnormal expression and function of the desmoglein was proved in many diseases,such as skin,mucous membrane and tumor related diseases.Drugs may have an important effect in the treatment of diseases by interfering with the expression and function of desmoglein.In this paper,the distribution of several subtypes of the family of desmosomes and their functions in the related diseases are reviewed,which may also provide some new clues for the new drug research on the target of desmogleins.

Objective To investigate the protective effects of Kudiezi (KDZ) Injection on cerebral ischemia reperfusion injury in rats and to explore its protective mechanism.Methods The rat model of middle cerebral artery occlusion (MCAO) was established by modified suture method,and cerebral blood flow was monitored using laser Doppler flowmetry (LDF).Male SD rats were randomly divided into control group,model group,Kudiezi Injection high and low dose groups.After ischemia-reperfusion for 24 h,the neurological scores were evaluated.After anesthesia,the blood samples and brain tissues were collected,and the expression of inflammatory was detected by Elisa.Western blotting was used to detect the expression of TLR-4 and NF-κB protein.Results Behavioral scores showed that neural function defect was serious in model group compared with control group (P ＜ 0.01).In model group,cerebral index and cerebral infarction area were significantly higher than those of the control group;After KDZ intervention,the symptoms of neurological deficit was alleviated (P ＜ 0.01),the cerebral index and cerebral infarction area of mode were decreased,and the neuronal necrosis was reduced.Kudiezi Injection could significantly reduce the cerebral homogenate and serum levels of TNF-α (P ＜ 0.05) and increase IL-10 level (P ＜ 0.05).Westem blotting showed that Kudiezi Injection could reduce the expression of TLR-4 and NF-κB protein (P＜0.05).Conclusion Kudiezi Injection has protective effect on cerebral ischemia reperfusion rats.After ischemia-reperfusion,Kudiezi Injection could reduce the levels of TNF-α and raise IL-10.Its mechanism may be associated to the down-regulation of TLR-4/NF-κB signaling pathway.