Objective To explore the infections of human cytomegalovirus (HCMV) in chronic hepatitis C patients and the hepatic impairment in chronic hepatitis C patients co-infected with HCMV.Methods HCMV-DNA was determined by fluorescence quantitative-PCR (FQ-PCR) in 95 patients with chronic hepatitis C (observation group) and 95 healthy controls(control group) and HCMV active infections were analyzed.HCV-RNA was determined by FQ-PCR in observation group,and the difference of HCMV-DNA positive rate between high HCV-RNA(＞ 104 copies/ml) and low HCV-RNA(≤ 104 copies/ml) was analyzed.Alanine aminotransferase (ALT),aspartate aminotransferase (AST) were determined by rate method in two groups and the hepatic impairment was analyzed.Results Twenty-five cases with positive HCMV-DNA in observation group,the positive rate was 26.3％(25/95).Five cases with positive HCMV-DNA in control group,the positive rate was 5.3％(5/95).There was significant difference between two groups for HCMV-DNA (x2 =14.29,P ＜0.01).Twenty-one cases with positive HCMV-DNA in 43 cases of high HCV-RNA patients,the positive rate was 48.8％(21/43).Four cases with positive HCMV-DNA in 52 cases of low HCV-RNA patients,the positive rate was 7.7％(4/52).There was significant difference between the two (x2 =19.90,P ＜ 0.01).ALT,AST in observation group was higher than that in control group (P ＜ 0.01).ALT,AST in chronic hepatitis C patients positive for HCMV-DNA was higher than that in chronic hepatitis C patients negative for HCMV-DNA significantly (P ＜ 0.01).Conclusions HCMV in chronic hepatitis C patients becomes active again and co-infects easily.When chronic hepatitis C patients co-infect HCMV actively,hepatic is further injured.

Objective To study the impact on pregnant outcome of reducing the number of embryos transferred from three to two in women at age less than 35 who received frozen-thawed embryo transfer (FET). Methods The analysis was performed on 90 FET cycles (77 infertile couples,less than 35 years old) with slow-freezing/rapid-thawing method, including 48 cycles with two embryos transferred and 42 cycles with three embryos transferred. The embryo survival rate, high quality embryo rate, clinical pregnancy rate, implantation rate and multiple pregnancies rate were analyzed. Results No significant differences in embryo survival rate (88.9% versus 88.1%), high quality embryo rate (89.6% versus 81.0%), clinical pregnancy rate (37.5% versus 42.9%), implantation rate (26.0% versus 18.3%) and multiple pregnancy rate (38.9% versus 16.7%) were observed between two and three embryos transferred group (all P > 0.05). However, there were 2 triple pregnancies in three embryos transferred group while none in two embryos transferred group. Conclusion Reducing the number of high quality embryos transferred from three to two in women at age of less than 35 years old who received FET,could decrease the incidence of triple pregnancy and keep the similar clinical pregnancy rate.

BACKGROUND: Cognitive impairments are often accompanied in patients with silent cerebral infarction (SCI) and depression, and there is still no reliable objective assessing method for the evaluation.OBJECTIVE: The purpose of this study was to probe into the value of auditory P300 in detecting the cognitive impairment in SCI patients with depression.DESIGN: A golden standard control study.SETTING: Department of Neurology, Xi 'an Central Hospital; Department of Neurology, the Second Hospital of Xi' an Jiao Tong University.PARTICIPANTS: Thirty-eight SCI patients were selected from the Outpatient Department of Neurology, the Second Hospital of Xi' an Jiao Tong University between January 2002 to May 2003, including 10 got dizziness,11 got poor concentration, 8 got headache, 5 got numbness of extremities and 4 got syncope. Among them, 26 were detected by CT and 12 by MRI.The patients'aged 45 to 63 years with mean age of (57.3±5.1) years, including 20 males and 18 females.METHODS: The existence of depressive symptoms was measured with the 17-item Hamilton depression scale (HAMD). In this scale, the total score ranged from 0 to 52, with higher score indicating more severe depressive symptoms, and the patient was considered as having depression if the HAMD score was ≥ 9. HAMD tests were administered by two psychiatrists,who were experienced in psychiatric disorders in stroke and other somatic diseases, and the two psychiatrists had a better coincidence (Kappa=0.86 -0.91). The auditory event related potential was detected with MYTO serial cerebra induced potential meter, while lying in a bed placed in a sound-attenuated room. Auditory stimuli were delivered binaurally through headphones. The subject was required to distinguish between the tones by noting the occurrence of the target and not to respond to the standard. All patients underwent a routine experimental test at first, and then a normal test was performed to obtain a better coincidence. Mini-mental state examination (MMSE) was conducted for the 38 patients by specialized training neurologist to assess the cognitive impairment, in which the total score ranged from 0 to 30, with lower score indicating greater impairment. The criteria of intellectual deterioration was defined on the basis of educational level:MMSE score of 17 for illiterate, 20 for primary school and 24 for middle school and above.MAIN OUTCOME MEASURES: The score of MMSE, latent period of P300 wave and amplitude of P300 wave were mainly observed.RESULTS: All the 38 cases were involved in the final statistical analyhave depressive symptoms (47%, depression group) and 20 had no depresindexes and score of MMSE between the SCI patients with and without depression: As compared with the SCI patients without depression, the P300 latent period was obviously prolonged, the P300 amplitude and MMSE score were markedly decreased in the SCI patients with depression [(383.3±31.1), (347.2±37.3) ms; (5.6±0.7), (9.9±1.6) μV; (15.2±1.1),and P300 indexes: In the SCI patients with depression, the P300 latent period had significant negative correlation with the MMSE score (r=-0.549, P ＜ 0.05), the P300 amplitude had significant positive correlation with the MMSE score (r =0.525, P ＜ 0.05). However, there was no correlation between the latent period and amplitude of P300 and the MMSE score in the SCI patients without depression (r=-0.349 to 0.065, P ＞ 0.05).CONCLUSION: Depression and cognitive impairments may have the same pathophysiological basis in SCI patients. P300 can be used as a referential and objective method to analyze the cognitive function in SCI patients with depression.

Objective To establish a molecular inversion probe ( MIP) method for detection of single base drug-resistance mutation in Hepatitis B virus ( HBV) gene.Methods The HBV wild type and YVDD mutant strain were isolated by Daping Hospital of the Third Military Medical University.The MIP was designed and applied to detect the HBV drug-resistance YVDD mutation in one case of wild type and one case of YVDD mutant HBV strain isolated previously.The results of MIP method were compared with that of sequencing to evaluate the detection accuracy.Results Thermal cycling single-base extension and connection reaction performed by Taq DNA Ligase and Ampligase DNA Ligase could ensure the specificity of the detection.The optimum probe concentration of MIP was 1 nmol/L.Through detection of the target gene with different DNA concentrations , the detection sensitivity of MIP was determined as 1 nmol/L.The results of MIP were consistent with that of sequencing method in detection of the clinical samples.Conclusion MIP is successfully used to detect single-base drug-resistance mutation in HBV gene.

Objective To explore the effects of luteolin on cognition function in pentylenetetrazol (PTZ)-induced epileptic rats and related mechanism.Methods Fifty male SD rats were randomly divided into a normal control group(n=8), a model group(n=12), and groups of 25, 50 mg/kg luteolin(both ofn=11), as well as 100 mg/kg luteolin group(n=8). Those rats were given different doses of luteolin (25, 50 and 100 mg/kg, daily, intragastric administration) for 36 consecutive days. Similarly, rats of the normal control group and the model group were given 0.5% sodium carboxymethyl cellulose suspension liquid via intragastric administration. Thirty minutes later, a model of epilepsy was induced using PTZ (40 mg/kg, daily) via intraperitoneal injection except the control group. Learning and memory of rats were evaluated by Morris water maze and novel objective recognition trials(including escape latency and recognition index). The levels of CaM and CaMPK were determined by ELISA methods, and expression of Ras proteins in the hippocampus were detected by Western Blot.Results Compared with the model group, luteolin treatment groups significantly shorten the escape latency(28.51 ± 3.84 s, 19.77 ± 5.41 s, 14.86 ± 2.76 svs. 37.08 ± 5.18 s) in the Morris water maze, and increased recognition index(18.77% ± 2.02%, 25.06% ± 4.32%, 31.92% ± 2.65%vs. 13.87% ± 2.14%) in the novel objection trial(P<0.05 orP<0.01). Meanwhile, CaM(140.33 ± 13.52 ng/L, 124.26 ± 9.97 ng/L, 113.52 ± 11.57 ng/Lvs. 158.36 ± 10.68 ng/L) and CaMPK(8.25 ± 1.37 ng/ml, 7.69 ± 0.84 ng/ml, 6.74 ± 0.93 ng/mlvs. 9.87 ± 1.02 ng/ml) were significantly decreased(P<0.05 orP<0.01). What’s more, the expression of Ras proteins(0.99 ± 0.08, 0.76 ± 0.07, 0.52 ± 0.07vs. 1.58 ± 0.12) was obviously decreased compared with the model group(P<0.05 orP<0.01).Conclusion Luteolin could effectively improve the cognition dysfunction of epileptic rats, and the mechanism might be relevant to regulate the CaM-CaMPK signaling pathway via down-regulation of CaM, CaMPK, as well as Ras protein.

OBJECTIVE To explore a more effective method of detecting sleep respiratory events in children. METHODS Thirty-eight children were tested with HypnoPTT and 31 with polysomnography (control). The test parameters and operative methods were compared. RESULTS In addition to the parameters comm.on to both HypnoPTT and polysomnography, inspiratory flow limitation and spontaneous microarousal can be measured using HypnoPTT and fewer electrodes were needed. CONCLUSION HypnoPTT is a convenient method characterized by less sleep disturbance and credible results, rendering it is especially suitable for pediatric cases. Pulse transit time is a useful parameter for diagnosing the sleep respiratory disease.

Objective To explore the inheritance characteristics of SCN1A gene in familial severe myoclome epilepsy in infancy.Methods The clinical information and blood of the patients and their relatives who had febrile seizure(FS)or epilepsy history were collected.Blood genome DNA were extracted.All exons of SeN1A gene were PCR amplified and screened with denaturing high Performance liquid chromatography(DHPLC)technology,and sequence analysis was performed.Results Fourteen SME patients had FS or epilepsy family history.Five were found positive history in first class relatives and 2 of them had inherited mutations of SCN1A(C.4284+2T>C and e.1216G>T):Other9 were found positive history in second class relatives and 2 of them had de novo mutations of SCN1A.Condusions SCN1A is the pathogenic gene for SME.The same muatation of SCN1A gene can be related to different clinical phenotypes.SME patients whose first class relatives with FS or epilepsy history should be taken as the focus of SCN1A inherited mutation screening.

Objective To prepare specific mouse monoclonal antibodies against Homo sapiens dynein,axonemal, heavy chain 2 (DNAH2). Methods Firstly, recombinant plasmid encoding His tagged immunogen, targeting N-terminal sequence of DNAH2 protein (1-300 aa), in E. coli was constructed. IPTG was used to induce the expression of His-immunogen, which was then purified and immunized in BALB/c mice. Hybridoma cells were obtained through the fusion between myeloma cells and splenocytes isolated from BALB/c mice. Finally, ELISA and Western blot assays were performed to screen the positive hybridoma. Results IPTG was used efficiently to induce the expression of DNAH2 immunogen in E. coli. DNAH2 protein bands were detected in screened positive hybridoma. Conclusion Mouse monoclonal anti-DNAH2 antibody is prepared successfully.

Objective To establish a homothermal and fast detecting method on pathogenic bacteria by combining recombinase-aid amplification (RAA) with molecular beacon.Methods The establishment of the methodology.Staphylococcus aureus specific primers were designed from the relative region of the staphylococcal protein A (SPA).Asymmetry amplification was optimized by adjusting the primer concentration ratios.The results of amplification and hybridization were visualized and analyzed by agarose gel electrophoresis and fluorescence detection.The sensitivity was identified by detecting dilute positive plasmids.And the specificity was determined using RAA method by detecting 72 pathogenic bacteria,including Staphylococcus aureus and other Staphylococcus spp.from the Department of Clinical Laboratory of Daping Hospital in December 2016.Besides,the Kappa analysis and the clinical diagnosis efficiency were investigated by analyzing 39 extra strains in the laboratory in December 2016.Results When the concentration ratio of restrictive and non-restrictive primer was 1:20,the yield efficiency of single-stranded DNA (ssDNA) reached the peak.And as for the hybridization efficiency,the asymmetry amplification was higher than symmetry amplification.Twenty copies/μl was proposed as the limits of detection by testing dilute plasmids.And the RAA hybridization method could distinguish Staphylococcus aureus with other Staphylococcus spp.Comparing with traditional detection methods with a Kappa index of 0.860,this method shows a good consistency.By analyzing the 111 bacteria,the sensitivity of the method is 92.5％ (37/40),the specificity is 97.2％ (69/71),the positive predictive value is 94.9％ (37/39),the negative predictive value is 95.8％ (69/72),the positive likelihood radio is 33.04,the negative likelihood radio is 0.077,the Youden index is 0.897 and the Kappa index is 0.902.Conclusion Through the combination of asymmetry recombinase-aid amplification optimization and molecular beacon probe,a new method of detecting bacteria DNA with RAA hybridization technique is established,providing the foundation for its clinical application.

Objective To understand the variation in response of Oncomelania hupensis to niclosamide. Methods Snails were collected from 37 sampling areas distributed in 10 provinces (municipalities) using random environmental sampling methods in accordance with the different types and categories of snail habitats. In laboratory the snails were immersed in solutions of niclosamide for 24 and 48 hours at 25℃. Results 1.0 mg/L niclosamide showed 100% killing effect on snails in 24 hours. The LC 50 concentrations for snails immersed for 24 hours ranged from 0.0320 to 0.1689 mg/L with a mean value of 0.0920 mg/L. 0.5 mg/L niclosamide showed 100% killing effect on snails in 48 hours. The LC 50 values for snails immersed for 48 hours ranged between 0.0299 and 0.1114 mg/L with a mean of 0.0627 mg/L. There is a significant difference in snail sensitivity to niclosamide between sampling areas. Conclusion The sensitivity to niclosamide varied in snails from different sampling fields, but the chemical in a concentration of 1.0 mg/L showed 100% effect of killing snails, which is consistent to the manual of schistosomiasis control.