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BACKGROUND Heterotopic pregnancy (HP) refers to the coexistence of ectopic pregnancy and intrauterine pregnancy. Salpingectomy is proposed as a pretreatment before in vitro fertilization and embryo transfer (IVF-ET) to reduce the risk of HP. HP after IVF-ET occurs in women who had already underwent bilateral salpingectomy, even though it is extremely rare. CASE SUMMARY A case of a 29-year-old woman with recurrent interstitial HP after IVF-ET following salpingectomy is presented. The main symptom was a sudden and worsening pelvic pain. Physical examinations revealed signs of peritoneal bleeding and irritation with stable vital signs. Transvaginal ultrasound showed a live intrauterine pregnancy and another live embryo with cardiac activity in the left cornu extending beyond the lateral edge of the uterus. Her hemoglobin concentration was 8.0 g/dL, and serum human chorionic gonadotropin value was 171116.9 mIU/mL. With the diagnosis of ruptured HP with internal bleeding, an emergency laparoscopic resection of left cornu was performed. The interstitial pregnancy was removed with caution to protect the intrauterine pregnancy. After the surgical treatment, the intrauterine pregnancy continued with no complications. A healthy baby was delivered by caesarean section at 39 wk. Outcomes of another three cases are further summarized. CONCLUSION Post-salpingectomy HP is a rare but challenging condition. Surgical treatment is preferred in the case with a viable intrauterine pregnancy.

Gastroesophageal reflux disease (GERD) often occurs in patients with obstructive sleep apnea (OSA). Although continuous positive airway pressure (CPAP) is considered to be the preferred treatment for OSA, the effect of CPAP therapy on reflux events remains controversial. In this study, we utilized meta-analysis to investigate whether or not CPAP treatment reduces the incidence of reflux. Two independent reviewers obtained the data sources from the database of PubMed, Elsevier, Cochrane library, and CNKI using search terms, and then filtered the target articles based on the inclusion and exclusion criteria. RevMan (version 5.3) and STATA (version 12.0) were used for data synthesis. The effect of CPAP treatment on GERD was studied by calculating the weighted mean difference (WMD) and standard deviation (SD) before and after CPAP treatment. Ten studies involving a total of 272 participants were included in this study. The results showed that the total of WMD before and after CPAP was − 17.68 (95% CI − 30.67 to − 4.69) for percentage time pH < 4, − 24.66 (95% CI − 36.15 to − 13.18) for the longest reflux duration, − 27.53 (95% CI − 49.53 to − 5.52) for number of reflux events, − 49.76 (95% CI − 60.18 to − 39.35) for DeMeester score, − 1.85 (95% CI − 3.00 to − 0.71) for reflux diseases questionnaire (RDQ) score, and − 8.95 (95% CI − 16.00 to − 1.89) for reflux symptom index (RSI). The subgroup analysis demonstrated that the improvement of reflux symptoms was more obvious with the extension of treatment time. This meta-analysis showed that CPAP treatment significantly reduces the incidence of reflux events in patients with OSA.

Introduction: To investigate the effects of paroxetine (PAR) on motor and cognitive function recovery in patients with non-depressed ischemic stroke (nD-AIS).Methods: One hundred sixty-seven patients hospitalized for non-depressed acute ischemic stroke were selected and divided into treatment (T) and control (C) groups using a random number table. All patients received conventional secondary ischemic stroke prevention and rehabilitation training; patients in Group T additionally received treatment with PAR (10 mg/day during week 1 and 20 mg/day thereafter) for 3 months. The follow-up observation lasted 6 months. The Fugl–Meyer motor scale (FMMS), Montreal cognitive assessment (MoCA), and Hamilton depression scale (HAMD) were used on D0, D15, D90, and D180 (T0, 1, 2, and 3, respectively; D180 = 90 days after treatment cessation) after study initiation, and scores were compared between the groups.Results: The FMMS and MoCA scores differed significantly between Groups T and C at T2 and T3 (p < .05); by contrast, these scores did not differ significantly between the groups at T1 (p > .05). Furthermore, the HAMD scores differed significantly between the two groups at T3 (p < .05), but not at T1 and T2 (p > .05).Conclusions: PAR treatment may improve motor and cognitive function recovery in patients with nD-AIS. Moreover, PAR may reduce the occurrence of depression after stroke.

a b s t r a c t To obtain the bioelectrodes with better stability, a novel construction of metal dry bioelectrode was designed to meet the specific application requirements of electrode performance for bioelec- tric impedance technique. The laser micromachining process was then employed to fabricate the microstructure arrays on the surface of dry bioelectrodes. Based on the material removal process of laser micromachining, the forming mechanism of surface microstructure arrays on the metal dry bio- electrode was further discussed. Using the two-electrode measurement method, the impedance values of electrode-skin for three types of bioelectrodes were recorded by measuring the left forearm position. Comparing to the conventional silver/silver chloride (Ag/AgCl) ECG electrode and metal planar bioelec- trode, the metal dry bioelectrode with surface microstructure arrays presented less than 50 � variations of contact impedance and better stability because of significant improvement of contact interface between electrode and skin, suggesting that it is an outstanding candidate for long-term clinical applications.

Objectives To investigate the association between toll-like receptor 9 ( TLR9) single nucleotide polymorphisms (SNPs) and human papillomavirus (HPV) infection among Chinese Han women with cervical cancer. Methods TLR9 –1486 and 2848 SNPs were investigated in patients with cervical cancer and controls using polymerase chain reaction (PCR)-restriction fragment length polymorphism. HPV16 E6 and E7 infections were assessed using PCR. Results Of 120 patients with cervical cancer and 100 controls, there was a significant association between TLR9 2848 SNP and cervical cancer risk, but there was no such association with TLR9 –1486 SNP. Frequency of the TLR9 2848 GA genotype was significantly higher in patients with cervical cancer than in controls. There was no statistically significant between-group difference in presence of HPV16 infection. Presence of HPV infection with TLR9 2848 (rs352140) GA/AA genotype increased the risk of cervical cancer 13.8-fold compared with the GG genotype. Conclusions The TLR9 2848 G/A polymorphism in Chinese Han women was associated with increased risk of cervical cancer in the presence of HPV16 infection. Further studies are necessary to uncover the functional aspect of this TLR9 2848 polymorphism.

To study the alternating current (AC) impedance properties of Ag/AgCl electrocardiograph (ECG) electrodes, the electrode pair was gel-to-gel connected, and then the electrical potential was recorded after a safe stimulating current passes through the electrode pair, so the AC impedance data of ECG electrodes were obtained. Varying the frequency and value of stimulating current, the detailed comparison and analysis of AC impedance properties of the electrodes were performed, and the stability was further characterized by using the continuous measurement within 24 h. The experimental results show that the AC impedance values of electrodes decreased, and then slightly increased with the increase of frequency of stimulating current. The minimum AC impedance value was obtained when the frequency was changed to 10 kHz. When the stimulating current increased, the AC impedance values of electrodes showed a slight decrease, but did not change significantly. Besides, the continuous measurement results show that the impedance value presented a significant increase in the initial 30 min, and then was stabilized in the following measuring process.

Background Hemoglobin is a precursor of antibacterial peptides. Our aim was to identify an antibacterial peptide in human endometrium. We tested the antimicrobial activities of hemoglobin and a derived peptide in vitro and in vivo in rats. Methods Samples (n = 3) were scraped from the surface of endometrium. Acid-soluble proteins underwent electrophoresis followed by gel overlay assay and reversed-phase high-performance liquid chromatography. Antibacterial activities were determined by agar radial diffusion assay. Purified peptides were further characterized by electrophoresis, mass spectrometry, N-terminal amino acid (AA) sequencing and protein structure analysis. A rat model was used to test the inhibitory activity of human hemoglobin on vaginal infection with Escherichia coli, using one experimental group (intravaginal hemoglobin, n = 9) and three control groups (n = 14). Vaginal histology was studied. Results The purified peptide exhibited potent antibacterial activities against E. coli ML-35P. The N-terminal AA sequence was F L S F P T T K T Y, identical to AA 32-41 of the human hemoglobin alpha chain, and it had the same mass (m/z = 6776.8) as the alpha chain 32-93 AA fragment, with at least three alpha-helices. Histology indicated that the hemoglobin group changed significantly compared with the matrix control group (no treatment after infection): the surface layer of stratified squamous epithelium was smoother, inflammatory cell infiltration was relieved in the lamina propria and congestion pattern was decreased. Conclusions These results suggest that erythrocytes from endometrium are another source of the antimicrobial molecules. Hemoglobin and its derived peptides may play a role in the host defense against pathogens in human vagina.

The monthly mean items of energy budget and air temperature, specific humidity and wind vector at 10m over China land and sea areas in 1998 were calculated based on Goddard Earth Observing Data Assimilation System, NASA, United States. The results of model calculation show that the largest surface net radiation of short wave appears in summer (July) in central areas of Xinjiang and Xizang, and reaches a value in high value central area of 275W/m2, and it appears in spring (April) in Yellow Sea and East China Sea with a its value about 250W/m2. The largest surface net radiation of long wave appears in summer (July) in northwestern China, with a value in central area of 125W/m2, and it appears in winter (January) in near China sea areas, with a value of 75–100W/m2. The latent heat flux in near China sea areas in winter (January) is higher than that in other months, its value in central area occurs in southwestern, northern, and northeastern China, and reaches 125W/m2. The calculated results of monthly mean energy budget illustrate that the net flux is transferred from ocean to atmosphere in winter (January) in Yellow Sea and East China Sea, and conversely in summer (July). The net flux value is positive in summer in Xinjiang and central area of Xizang Plateau. The comprehensive analysis for distributions of temperature, humidity and wind vector shows that the water vapour is transported distinctly in summer from South China Sea to eastern China area and from Bay of Bengal to India subcontinent. The water vapour in southwestern area of Xizang comes from its southwestern direction.

LRP2 (also called megalin) plays a potential key role in the pathogenesis of Alzheimer's disease (AD). Recently, one genome-wide association study has revealed that the rs3755166 (G/A) polymorphism located in the LRP2 promoter is associated with development of AD in Caucasians, while there are no studies on the association LRP2 of with AD risk in Asians.To evaluate the relationship between the rs3755166 polymorphism of the LRP2 gene and AD in the ethnic Chinese Han, we conducted a case-control study (n=361, age50) to determine the prevalence of one common single-nucleotide polymorphism (SNP) of LRP2 (rs3755166) in patients with AD in Chinese population of Mainland China, and clarified whether this polymorphism is a risk factor for AD.The prevalence of the minor allele (A) in the rs3755166 polymorphism was significantly different in AD patients and control subjects (P0.05). The rs3755166 polymorphism was associated with AD in the ethnic Chinese Han (OR=1.378, 95% CI: 1.017-1.867, P=0.039), and the results were not influenced by age, gender, or APOE status (P=0.441, P=0.94, P=0.432, respectively).Our data revealed the allele (A) of the rs3755166 polymorphism within LRP2 gene may contribute to AD risk in the Chinese Han Population.

To investigate the effect of Helicobacter Pylori lipopolysaccharide (Hp-LPS) on expression of Gli and Ptch-1 proteins in sonic hedgehog (Shh) signaling pathway of gastric mucosa GES-1 cells.The LPS was extracted from Hp by hot phenol water method, and then the concentration of LPS was detected by the kinetic turbidimetric assay. GES-1 cells were stimulated by different concentrations of Hp-LPS (0, 1, 10, 20, 30 and 40 μg/ml). The inhibition rates of cell growth were measured by MTT assay after treated with Hp-LPS for 24 h. The expression of Gli and Ptch-1 proteins were determined by Western Blot.MTT assay showed that the inhibition rates of GES-1 cell growth after treatment by different concentrations of Hp-LPS (1, 10, 20, 30 and 40μg/ml) were 25.8% ± 2.7%, 34.2% ± 3.1 %, 46.3% 3.4%, 60.8% ± 2.1% and 82.9% ± 2.8% respectively (r=0.985, P0.001). Western blot showed that the expressions of Gli and Ptch-1 proteins were decreased after Hp-LPS treatment (0, 1, 10, 20, 30 and 40 μg/ml): the relative expression values of Gli were 1.286 ± 0.180, 0.963 ± 0.067, 0.850 ± 0.085, 0.566 ± 0.058, 0.549 ± 0.056 and 0.377 ± 0.047, respectively (r=-0.945, P0.001); those of Ptch-1 were 1.688 ± 0.088, 1.466 ± 0.061, 1.170 ± 0.065, 1.042 ± 0.064, 0.648 ± 0.057 and 0.482 ± 0.074, respectively (r=-0.985, P0.001).Hp-LPS can decrease the related protein expression of Shh signaling pathway, which indicates that Hp may interfere with the function of Shh signaling pathway in gastric mucosa via the effect of its LPS.

Serotonin selective reuptake inhibitors (SSRIs) have been widely used as first-line drugs in the treatment of a range of depressive and anxiety disorders. Recently, clinical studies found that this class of agents also shows significant efficacy in promoting neurogenesis, neuroplasticity and neurorecovery following stroke. Here, we attempt to elucidate molecular mechanism and biological implication underlying the SSRI-mediated neurorecovery. In the procedure, a comprehensive protein-drug interactome (PDI) was constructed for various SSRIs and their major metabolites as well as a group of control drugs across a large panel of human neuroproteins via a high-throughput molecular docking approach. The obtained PDI was then analyzed at systematic level to extract unexpected targets for SSRIs/metabolites. Biological network analysis and gene ontology (GO) enrichment solidified that the inferred targets have high potential to be directly or indirectly involved in diverse neural events, and further molecular dynamics (MD) simulation and post molecular mechanics-Poisson Boltzmann/surface area (MM-PB/SA) characterization revealed a stable complex architecture and high-affinity interaction between the targets and SSRIs/metabolites. Specifically, two human proteins, i.e. neurogenic locus notch homolog protein 1 (NOTCH 1) and Rho-associated protein kinase 1 (ROCK 1), were suggested as promising regulators in the SSRI-mediated neurorecovery, which can be targeted efficiently by fluoxetine and paroxetine, respectively, as well as other SSRIs and metabolites.

Background C-src is an evolutionarily conserved proto-oncogene that regulates cell proliferation, differentiation and apoptosis. In our previous studies, we have reported that another proto-oncogene, c-erbB 2 , plays an important role in primordial follicle activation and development. We also found that c-src was expressed in mammalian ovaries, but its functions in primordial follicle activation remain unclear. The objective of this study is to investigate the role and mechanism of c-src during the growth of primordial follicles. Methods Ovaries from 2-day-old rats were cultured in vitro for 8 days. Three c-src-targeting and one negative control siRNA were designed and used in the present study. PCR, Western blotting and primordial follicle development were assessed for the silencing efficiency of the lentivirus c-src siRNA and its effect on primordial follicle onset. The expression of c-src mRNA and protein in primordial follicle growth were examined using the PCR method and immunohistochemical staining. Furthermore, the MAPK inhibitor PD98059, the PKC inhibitor Calphostin and the PI3K inhibitor LY294002 were used to explore the possible signaling pathways of c-src in primordial folliculogenesis. Results The results showed that Src protein was distributed in the ooplasmic membrane and the granulosa cell membrane in the primordial follicles, and c-src expression level increased with the growth of primordial follicle. The c-src -targeting lentivirus siRNAs had a silencing effect on c-src mRNA and protein expression. Eight days after transfection of rat ovaries with c-src siRNA, the GFP fluorescence in frozen ovarian sections was clearly discernible under a fluorescence microscope, and its relative expression level was 5-fold higher than that in the control group. Furthermore, the c-src-targeting lentivirus siRNAs lowered its relative expression level 1.96 times. We also found that the development of cultured primordial follicles was completely arrested after c-src siRNA knockdown of c-src expression. Furthermore, our studies demonstrated that folliculogenesis onset was inhibited by Calphostin, PD98059 or LY294002 treatment,but none of them down-regulated c-src expression. In contrast, the expression levels of p-PKC, p-ERK1/2 and p-PI3K in the follicles were clearly decreased by c-src siRNA transfection. Correspondingly, both Calphostin and LY294002 treatment resulted in a decrease in the p-PKC level in follicles, but no change was observed in the PD98059 group. Finally, LY294002 treatment decreased the p-PI3K expression level in the follicles, but no changes were observed in the PD98059 and Calphostin groups. Conclusions C-src plays an important role in regulating primordial follicle activation and growth via the PI3K-PKC- ERK1/2 pathway.

Japanese encephalitis virus (JEV), a mosquito-borne zoonotic pathogen, is one of the major causes of viral encephalitis worldwide. Previous phylogenetic studies based on the envelope protein indicated that there are four genotypes, and surveillance data suggest that genotype I is gradually replacing genotype III as the dominant strain. Here we report an evolutionary analysis based on 98 full-length genome sequences of JEV, including 67 new samples isolated from humans, pigs, mosquitoes, midges. and bats in affected areas. To investigate the relationships between the genotypes and the significance of genotype I in recent epidemics, we estimated evolutionary rates, ages of common ancestors, and population demographics. Our results indicate that the genotypes diverged in the order IV, III, II, and I and that the genetic diversity of genotype III has decreased rapidly while that of genotype I has increased gradually, consistent with its emergence as the dominant genotype.

PURPOSE To assess genetic variations of GAB2 as a risk factor for developing Alzheimer disease (AD). DESIGN AND METHODS A case-control study (n=310; age>50 y) was conducted to determine the prevalence of 5 single nucleotide polymorphisms (SNPs) of GAB2 (rs2373115, rs1385600, rs4945261, rs7101429, and rs7115850) in patients with AD in Chinese population of mainland China, and was investigated whether these polymorphisms are risk factors for AD. RESULTS Our results supported a possible implication of 3 tested SNPs of GAB2 (rs4945261, rs7101429, and rs7115850) in AD in the ethnic Chinese Han, of which the maximal significance of association was at SNP rs7101429 C allele (P=4.0×10; odds ratio=2.0; 95% confidence interval, 1.4-2.8), and this observed association was not affected by APOEe4 genotype. In the haplotypes analysis, the minor alleles of the 3 tested SNPs were composed of a TCG haplotype, which had a significant difference in haplotype distribution between the 2 groups (P=3.4×10; odds ratio=8.32; 95% confidence interval, 4.57-15.14). CONCLUSIONS Our findings implicate an association between genetic variations of GAB2 and AD in Han Chinese, and the minor alleles of the 3 tested SNPs (rs4945261, rs7101429, and rs7115850) might increase the risk of AD.

To study the expression and possible roles of proto-oncogene c-erbB2 during the initiation growth of primordial follicles.Ovaries were collected from 2-day-old SD rats and cultured in the Waymouth culture system. In-situ hybridization, RT-PCR and immunohistochemistry were performed to assess the expressions of c-erbB2 mRNA and protein during the initiation growth of primordial follicles and after the effect of EGF. Western blot was used to observe the PCNA, p-ERK1/2 contents and correlation analysis was used to study the correlation relationship between contents of p-ERK1/2 and expressions of c-erbB2 mRNA at the same time of the primordial follicles growth.PCNA protein levels appeared to be more intense during the initiation growth of primordial follicles, EGF could promote the proliferation and differentiation of the primordial follicles. c-erbB2 mRNA existed in the oocytes endochylema and ErbB2 existed in the oocytes membrane, the expressions of c-erbB2 mRNA and ErbB2 appeared to be more intense when primordial follicles were cultured for 8 d than cultured for 0 d in the Waymouth culture system and were further increased with 50 ng/ml EGF for 4 d and 8 d. The same results were observed by RT-PCR, too. p-ERK1/2 protein levels were consistent with the changes of c-erbB2 mRNA and protein. Furthermore, Spearman rank correlation analysis showed there was a significant positive correlation relationship between the changes of p-ERK1/2 and the changes of c-erbB2 mRNA during the primordial follicles growth and after the effect of EGF (rs = 0.900, P0.05).It was suggested that proto-oncogene c-erbB2 may be play an important role during the initiation growth of primordial follicles with EGF, and it is indirectly suggested that c-erbB2 promotes the development of the primordial follicles via ERK-MAPK signal transduction.

The growth factor receptor-bound protein 2 (Grb2)-associated binder (Gab) proteins are intracellular scaffolding/docking molecules, and participate in multiple signaling pathways, usually acting as the downstream effector of protein-tyrosine kinases (PTKs)-triggered signal transduction pathway. When phosphorylated by PTKs, Gab proteins can recruit several signaling molecules (p85, SHP2, and Crk), and subsequently activate multiple transmitting signals that are critical for cell growth, survival, differentiation and apoptosis. Recently, it has been reported that Gab2 polymorphism is associated with the increase in the risk of Alzheimer's disease (AD) and is involved in the pathogenesis of AD. This review mainly focuses on the structure and function of Gab2 protein and its role in the pathogenesis of AD.

For constructing the high-throughput platform of sequencing the JEV whole genome, the two systems of multiplex primers for genotype I and III should be designed and used for detected the whole genome of genotype I and III JEV in the research.The two systems of JEV genotype-specific primers was designed based on the reference sequence of all the available genotype I and III JEV genome sequence on GenBank, then, they were used to amply and sequence the 121 JEV strains isolated in China contains 63 GIII JEV and 58 GI JEV.The self-designed genotype-specific primers for genotype I and genotype III JEV were 16 pairs and 17 pairs respectively, which were used for detecting the whole genome of 121 JEV. The average quality value for GI JEV is 40.037. The average quality value for GIII JEV is 40.857.The two systems of JEV genotype-specific primers could sequenced the genotype I and III JEV qualified and specific. It is the basis of the high throughput platform of sequencing the JEV whole genome.

To sequence and analyze the whole genome of Japanese encephalitis virus (JEV) isolated from mosquitoes in Liaoning province in 2008.Using RT-PCR to amplify fragments with genome sequencing primer. The full-length genome was obtained by sequencing and splicing. The differentiation analysis for nucleotides, deduced amino acid sequence and phylogenetic tree was performed by the software of Clustal X (1.83), ATGC (V4), DNAStar, GENEDOC (3.2) and Mega (4.0).The whole genome of strain LN0828 possesses 10 965 nucleotides. An open reading frame from 97 to 10 392 including 10 296 nucleotides is capable of coding for a 3432 amino acid polyprotein. Comparison of strain LN0828 genomic sequence with those of 32 JEV isolates in GenBank showed that nucleotide sequence divergence ranges from 1.6% to 16.4%, which resulted in amino acid sequence divergence from 0.3% to 5.1%. In comparison with live attenuated vaccine stain SA14-14-2 in open reading frame, strain LN0828 has a total of 1186 nucleotide substitutions, 86 amino acid divergences. Based on phylogenetic analysis, the strain LN0828 belongs to the genotype I JEV.The whole genome of strain LN0828 is close to those of isolates from Liaoning in 2002 and 2007, which were grouped into genotype I JEV.

To analyze the molecular characteristics of Japanese encephalitis virus (JEV) isolated in Northwestern Yunnan province, and to clarify the differences between the strains isolated in Northwestern and other parts of Yunnan province.PrM, E and 3' untranslated region nucleotide acid sequences of the isolates were amplified by RT-PCR and then sequenced. Sequence alignment and phylogenetic analysis were performed by using Clustal 1.8X, DNASTAR, GENEDOC and Mega 3.1 programs.12 of the 13 isolates of JEV obtained in Northwestern Yunnan were identified as genotype I, only one strain was genotype III of JEV. The 12 strains of genotype I were clustered in different branches with other isolates obtained in other parts of Yunnan province. Data from sequence analysis on E gene found that the nucleotide identity was 0.2%-13.9% between the Northwestern isolates and other Yunnan strains. There were two kinds of nucleotides deletion patterns at 3' untranslated region with three and one deletions was found after termination codon in genotype I and III isolates, respectively.There were two genotypes of I and III in 13 strains of JEV in this study and genotype I isolates were predominant (12/13). There were no apparent differences in E gene sequence between isolates obtained in the Northwestern and other parts of Yunnan. Three deletions were found in 3' untranslated region in genotypes I isolates and one deletion was in genotypes III.

To sequence and analyze the whole genome of Japanese encephalitis virus (JEV) strain named 47 which was isolated from patient's cerebrospinal fluid sample in Heilongjiang province in 1950.RNA was extracted from the recovery strain 47 and amplified with self-designed JEV genome sequencing primers. The differentiation analysis for nucleotides and coding amino acids and phylogenetic analysis were performed by the software of DNAStar, Modeltest, and Phylip.The whole genome of strain 47 has 10,977 nucleotides. An open reading frame from 95 to 10,391 including 10,296 nucleotides is capable of coding a 3432 amino acid polyprotein. The nucleotide difference between strain 47 and 5 vaccine strains is 2.4%-4.4%, the amino acid difference between strain 47 and 5 vaccine strains is 0.3%-1.1%. The best evolution model for the whole genome is GTR + I + G. Based on the phylogenetic analysis, strain 47 belongs to the genotype III JEV.Strain 47 is highly conserved on whole genome nucleotide and amino acid sequence. And it is belongs to the genotype III JEV.

ARLTS1 has been identified in chromosome 13q14 as a tumor suppressor gene of the adenosine diphosphate-ribosylation factor family with pro-apoptotic characteristics. The ARLTS1 mutation Trp149Stop and Cys148Arg have been shown to be associated with familial cancers, but limited information is available regarding the impact of ARLTS1 variants on familial ovarian cancer (OC). The aim of this study was to evaluate the ARLTS1 genetic variants associated with familial OC risk in China. We genotyped 85 OC patients with family ovarian/breast history, 80 sporadic OC patients, and 120 controls from general population by denaturing high-performance liquid chromatography screening analysis followed by direct sequencing of the conspicuous polymerase chain reaction products. ARLTS1 Cys148Arg revealed a significant association with an increased risk of familial OC compared with both sporadic cases and controls in a dose-dependent manner (P = 0.0031 and 0.012, respectively). In the clinical-pathological study, our results support previous data in demonstrating that familial OC was associated with younger age at diagnosis (49.7 years vs 53.3 years; P = 0.014), higher proportion of tumors of advanced stages (81.2% vs 67.5%; P = 0.033), and higher rates of serous adenocarcinomas (76.4% vs 53.8%; P = 0.028) compared with sporadic OC cases. To investigate the association between genetic variants of ARLTS1 and the clinical-pathological characteristics of familial OC, we identified a significantly higher proportion of serous adenocarcinoma (55/67, 82.1%) and higher rates of advanced stage tumors (88.1% vs 55.6%; P = 0.004) in ARLTS1 Cys148Arg carriers. We showed a significantly increased risk of familial OC for ARLTS1 Cys148Arg variant, which indicate that ARLTS1 may play a role in familial OC.

To investigate the susceptibility of cervical carcinoma cells (HPV16+) to CTL lysis affected by rSIFN-co, consensus Interferon (Infergen), IFNalpha-2b and DDP.After CaSki cervical cancer cells were induced by rSIFN-co, Infergen, IFNalpha-2b and DDP at the concentration of 0.156 microg/mL, 0.625 microg/mL, 2.500 microg/mL for 72 hours, CaSki cells which had been induced were effected by CTL, the cytotoxicity was determined and calculated by MTT assay. The expression intensity of adhesion molecules on Caski cell such as CD54 and CD40 was also determined by flow cytometry.The susceptibility of CaSki cell to CTL lysis under the stimulation of rSIFN-co was better than Infergen, IFNalpha-2b and DDP induced. The expression of CD54 and CD40 on cervical cancer cell was also increased. And this effect had positive correlation to the drug concentration.rSIFN-co can increase the expression of CD54 and CD40 on the cervical cancer cell surface, and increase the susceptibility of CaSki cell to specific effective cell lysis in a dose-dependent manner. The effect of rSIFN-co is better than same type interferon, general I type interferon and chemotherapeutic drug induced.

To explore the effect of metastasis suppressor gene KAI1 on the proliferation and invasive ability of cervical cancer cell line CaSki.pCMV-KAI1 cDNA plasmid was transferred into cervical carcinoma cell line CaSki by liposome, which had low level of endogenous KAI1 expression. The expressions of KAI1 protein and mRNA were determined by immunohistochemistry and real-time fluorescence quantitative PCR (RT-PCR), the proliferation of KAI1-transfected CaSki cells was investigated by MTT assay and the invasive ability of these cells was evaluated by in vitro invasion assays.After the transfection of pCMV-KAI1 cDNA, the level of KAI1 mRNA and protein expression in CaSki cell were increased (P0.05), while the cell proliferation was suppresssed, and the migrative ability of passing through the membrane filte also decreased evidently (P0.05).The KAI1 metastasis suppressor gene suppressed the ability of proliferation and invasion of cervical cancer cell CaSki in vitro.

To isolate human hemoglobin and its fragrments, compare their antimicrobial activity in vitro and pilot study of their antimicrobial activity in vivo.The alpha and beta chains of hemoglobin were separated by cation exchange chromatography and gel chromatography; The alpha and beta chains were cleaved by cyanogens bromide respectively. The cleaved fragments were purified by reverse phase high performance liquid chromatography. Antimicrobial activity of hemoglobin and its fragments was determined by agrose radial diffusion assay. After establishment of E. coli vaginal infection model, the rats were randomized into the experimental group (hemoglobin group) and the control group. The histologically pathological section was observed.Hemoglobin, alpha/beta chain and their fragments had similar antibacterial activities in vitro, which were mainly against Gram-negative bacteria E. coli; except alpha1-32 had a comparatively lower activity. Antimicrobial activity in vivo: a comparison of the hemoglobin group with the matrix control group (no treatment after infection), the surface layer of vaginal stratified squamous epithelium was smoother, inflammatory cells were significantly reduced in the lamina propria and congestion was obviously decreased.Human hemoglobin and its fragments had antibacterial activity in vitro, hemoglobin might relieve the inflammation of E. coil vaginal infection in rats moreover.

To investigate the expression of metastasis suppressor gene KAI1 in cervical carcinoma and the impact of human papillomavirus 16 E6, E7, 18 E6/E7 infection on the expression of KAI1.The expressions of KAI1 protein in the formalin-fixed, paraffin-embedded specimens of 20 normal cervical epthelium, 15 cervical in situ carcinoma and 70 primary invasive cervical carcinoma were detected by immunohistochemistry SP. Polymerase chain reaction (PCR) tests were also undertaken to detect the HPV16 E6, E7 and HPV18 E6/E7 DNA.The expression of KAI1 protein was down-regulated in the invasive carcinoma and in situ carcinoma compared with the controls (P0.05). No significant difference was found in the expression of KAI1 protein between invasive carcinoma and in situ carcinoma. The infections of HPV16 E6, E7 and HPV18 E6/E7 were found in 67.1%, 54.3% and 12.9% of the invasive carcinoma, respectively. However, there was no correlation between the expression of KAI1 and the infections of HPV16 E6, E7 and HPV18 E6/E7.The expression of KAI1 protein is down-regulated in cervical carcinoma, which is not associated with the infection of HPV16 E6, E7 and 18 E6/E7.

To explore the possibility of using lymphokine activated killer (LAK) from cord blood as an adoptive immunotherapy for ovarian cancer.The nude mouse models with human ovarian cancer were treated with the LAK from cord blood and compared with those treated with the LAK from the pheripheral blood.The LAK from the cord blood inhibited the growth of human ovarian cancer. The average size of the tumor was 6.0 mm3 23 days after the cord blood LAK treatment, with an average weight of (0.674 +/- 0.45 g).Cord blood can be a source of adoptive cellular immunotherapy for the treatment of ovarian cancer.

To isolate and purify antimicrobial peptides from human uterus mucus.Acid-soluble extract was obtained from the specimens of endometrium mucus from 3 hysteromyomas patients during total hysterectomy. Acidic urea-polyacrylamide gel electrophoresis was used to analyze the acidic extract. The corresponding antimicrobial band HUP was further isolated and purified by electrophoretic elution and reversed-phase high-performance liquid chromatography (RP-HPLC). The antimicrobial activity of the fractions was analyzed by agarose radial diffusion assay. The molecular weight was determined by Tricine-SDS-PAGE. According to the results of the N-terminal sequencing and Mass Spectrometry analysis, the amino acid sequences of the purified molecules were deduced. The deduced amino acid sequence of the antibacterial fragment was further analyzed by ExPASy and OMIGA softwares.An antibacterial peptide named HUP-39 was purified from the human uterine mucus with a molecular weight of 6.777 Ku. N-terminal sequencing and Mass Spectrometry analysis suggested that this antimicrobial peptide should be hHEM-alpha 33-95 amino acid fragment. ExPASy and OMIGA analysis showed that it contained 3alpha-helical transmembrane domains and its pI was 8.38. The fragment was mainly against Escherichia coli ML-35p, E. coli ATCC 25 922, and the clinically isolated strain E. coli 54 080.An antimicrobial peptide has been isolated and purified from human uterine mucus, a hHEM-alpha 33-95 amino acid fragment. The antimicrobial molecule in the uterine mucus originates not only from epithelial cells and leucocytes, but also from erythrocytes.

To isolate and purify antibacterial polypeptides from human cervical mucus.Human cervical mucus was collected from human healthy subjects and acid-soluble extract was obtained by solving the mucus with 5% acetic acid in the presence of protease inhibitors. The antibacterial components were identified by Agar radial diffusion assay and gel overlay technique. For further purification, Preparative acid-urea gel electrophoresis and Reverse Phase HPLC were performed. The N-terminal sequencing and degenerate PCR-directered cDNA cloning were performed. The E. coli-based recombinant product was prepared and its antibacterial property was determined by minimal inhibitory concentration and minimal bactericidal concentration.A purified antibacterial polypeptide was obtained. Agar radial diffusion assay showed that the purified polypeptide had antibacterial activities against E. coli ML-35P and Pseudomanas aeruginosa ATCC 27853. The N-terminal amino acid sequence and its full length of cDNA were identical to High Mobility Group Chromosal protein N2 (HMGN2). The purified recombinant HMGN2 was obtained. Its MIC against E. coli ML-35p and P. aeruginosa ATCC27853 were 10.42 microg/ml +/- 3.13 microg/ml and 27.78 microg/ml +/- 8.33 microg/ml respectively, which were equal to human neutrophil defensins HNP, and the MBC were 20.83 microg/ml +/- 6.25 microg/ml and 55.56 microg/ml +/- 16.67 microg/ml respectively.HMGN2 may be another antibacterial effecter in the defense mechanisms of human cervical mucus.

To identify a new antibactrial polypeptide HCP-1 isolated from human cervical mucus.HCP-1 was isolated and purified from human cervical mucus, and N-terminal sequence of HCP-1 was determined. A degenerate primer was designed according to CodeHop methods, and an "anchor-oliga-dT primer" was used for the synthesis of cDNA. Using the degenerate primer and anchor-primer, cDNAs were amplified by PCR. The PCR products were cloned, sequenced, and analyzed by biological software.N-terminat sequence of HCP-1 was PKRKAEGDAK. The full length of HCP-1 cDNA was isolated and of which the sequence was the same as HMG-17. OMIGA software analysis indicated that this molecule contained an alpha-helix region.The new antibacterial polypeptide isolated from human cervical mucus is HMG-17. It may play a role in the human cervical mucus and the alpha-helix domain may be its antibacterial activity region.

A subtracted cDNA library specific to osmotic stress of Haloxylon ammodendron (Mey.) Bge seedlings was constructed by suppression subtractive hybridization (SSH) and T/A cloning. SSH was performed between two groups of H. ammodendron seedlings, one was cultivated in Hoagland (H) solution as a driver and the other group was treated with osmotic stress of the Hoagland solution by the addition of 400 mM mannitol (M), as a tester. The library consisted of about 400 recombinant clones, with the average size being of 500 bp, ranging from 300 bp to 1500 bp. Using a PCR-select differential screening kit, 100 recombinant clones were randomly chosen from the subtracted cDNA library and hybridized with forward, reverse subtracted and unsubtracted probes for two rounds. As a result, 21 positive clones specific to osmotic stress were obtained and some of them were verified by Northern blot analysis. The sequencing analysis of 6 positive clones and the following homology comparison to GenBank [blastx] non-redundant databases characterized that two sequences obtained in this experiment may contribute to novel drought-related genes.