Objective To establish a method for the rapid detection of hepatitis E virus (HEV)from serum samples based on fluorescence quantitative PCR.Methods (1 )One-hundred HEV sequences including our country popular three major genotypes were obtained from the GeneBank with the Vector NTI software.The proper sequence was selected to design and synthesize the primers of the fluorescence quantitation and the Taqman probe.(2)The amplification region PCR fragment was transcribed in vitro to synthesize cRNA standard,at the same time the trace serum virus lysate was introduced into a universal real-time TaqMan PCR assay.(3)10 clinical serum samples were collected from the patients with clinical hepatitis E and detected by using the established method for further verifying this method.Results This detection technique could effectively detect the serum samples in the pa-tients with genotype I and genotype IV hepatitis E positive,while the serum detection in the patients with other virus infectious dis-eases had the negative results,which verified that this RT-PCR detection technique had higher specificity and good reliability.The detection results from 10 clinical serum samples further verified that this method was rapid,convenient and sensitive with good re-peatability.Conclusion A fluorescence quantitative RT-PCR detection technique suitable for detecting main genotypes of HEV in China population is established,which can meet the demand of early and rapid diagnosis for HEV.

Objective To primarily study the possible biological significance of yon Willebrand factor (vWF) expression at the fracture site in the mechanism of fracture healing. Methods A total of 28 male SD rats were selected to set up femoral fracture models. Then, tissue samples from fracture site were randomly taken at days 2, 3, 5, 7, 10, 14 and 21 after establishment of the models, with 4 rats at each time point, vWF expression at fracture site and the relationship of vWF expression with osteoblasts and chondroblasts were observed by using computer image analysis system. The biological significance of vWF expression at fracture site in mechanism of fracture healing were analysed based on the results of vWF expression of in vitro cultured rat osteoblasts. Results The expression of vWF at every time point was positive and distributed disproportionally irregular at bone trabecula and soft tissue of the fracture site. vWF expressed positively in the extracellular matrix around the chondroblasts. Conclusions During the course of fracture healing, there may be vWF-platelet activation pathway that can regulate differentiation and proliferation of local osteoblasts and chondrocytes and activate platelets secreting growth factors such as PDGF, IGF-1, EGF and TGF-a, as creates a sound microenvironment for fracture healing.

OBJECTIVETo explore the expression of Ghrelin and high mobility group protein B1 (HMGB1) in the serum and the intestinal tissue of sepsis model rats, and to evaluate the effect of electro-acupuncture (EA) at Zusanli (ST36) on the expression of HMGB1 and Ghrelin.

METHODSForty-eight male Wistar rats were randomly divided into four groups, i.e., the sham-operation (sham), the cecal ligation and puncture group (CLP), the CLP + EA at Zusanli (ST36) group (EA), and the CLP + Ghrelin receptor blocking agent + EA group (GHSRA), 12 in each group. A sepsis rat model was prepared by CLP. The incision of the abdominal wall was immediately sutured along the ventral midline for rats in the Sham group. In the EA group EA at Zusanli (ST36) was performed 20 min after CLP surgery with the constant voltage (2 - 100 Hz, 2 mA) for 30 min. In the GHSRA group, Ghrelin receptor blocking agent, [D-Arg1, D-Phe5, D-Trp79, Leu11]-substance P (700 nmol/kg), was administered through intravenous injection immediately after CLP, and 20 min later, EA at Zusanli (ST36) was performed in the same way as for rats in the EA group. Blood samples were withdrawn 12 h after CLP. The serum levels of Ghrelin and HMGB1 were detected using ELISA. Ghrelin expressions and the number of Ghrelin immunopositive cell in the jejunum were determined by immunohistochemistry. HMGB1 contents of the jejunum tissue were detected by Western blotting.

RESULTSCompared with the Sham group, the number of serum immunopositive cells and the expression of HMGB1 in the jejunum tissue significantly increased and levels of Ghrelin and the expression rate of immunopositive cells significantly decreased in the CLP group (P < 0.05). Compared with the CLP group, the number of serum immunopositive cells and the expression of HMGB1 in the jejunum tissue significantly decreased, but levels of Ghrelin and the expression rate of immunopositive cells significantly increased in the EA group (P < 0.05). Compared with the EA group, the number of serum immunopositive cells and the expression of HMGB1 in the jejunum tissue significantly increased in the GHSRA group (P < 0.05), but there was no statistical difference in levels of Ghrelin between the two groups (P > 0.05). The serum level of HMGB1 was negatively correlated with Ghrelin in the Sham group, the CLP group, and the EA group (r = -0. 528, P < 0.01).

CONCLUSIONSEA at Zusanli (ST36) could inhibit the expression of HMGB1 in the jejunum of septic rats, and promote the expression of Ghrelin. The expression of HMGB1 was inhibited by Ghrelin receptor blocking agent, which suggested that the anti-inflammation of EA at Zusanli (ST36) might be associated with Ghrelin.

Animals ; Disease Models, Animal ; Electroacupuncture ; Ghrelin ; metabolism ; HMGB1 Protein ; metabolism ; Jejunum ; metabolism ; Male ; Rats ; Rats, Wistar ; Sepsis ; metabolism

The postpartum depression outcome and the effect of psychological intervention were studied in order to reduce the occurrence and development of the postpartum depression. A survey of 4000 women within 4-6 weeks postpartum in 80 communities in Shenzhen, China was performed using random cluster sampling method. By employing Edinburgh Postnatal Depression Scale (EPDS) as a screening tool, the positive women (defined as EPDS ≥10) were randomly divided into intervention group and control group at a ratio of 1:2. The women in the intervention group were treated by means of mailing postpartum depression prevention and treatment knowledge manual, face-to-face counseling, and telephone psychological counseling interventions aiming at individual risk factors, while those in the control group were treated with conventional methods. EPDS scores were assessed in these two groups again at 6th month postpartum. Totally, 3907 valid questionnaires were obtained. All the 771 positive women were divided into two groups: 257 in the intervention group, and 514 in the control group. At 6th month postpartum, the EPDS scores in the intervention group were decreased significantly, from baseline stage (12.84±3.02) to end stage (3.05±2.93), while EPDS scores in the control group were reduced from 12.44±2.78 to 6.94±4.02. There were significant differences in the EPDS scores at end stage between the two groups (t=13.059, P<0.001). Psychological intervention can reduce postpartum depression, with better maternal compliance. It is feasible and necessary to establish postpartum depression screening and psychological intervention model in community-hospital and include the postpartum depression screening, intervention, and follow-up into the conventional healthcare.

OBJECTIVETo evaluate the accuracy of the Helicobacter pylori (H. pylori) stool antigen (HpSA) test and ImmunoCard STAT HpSA in the primary diagnosis of H. pylori infection.

METHODSWe searched Medline (1966-2007.4), EMbase (1985-2007.4), Chinese Journals Full-text Database (CJFD) (1994-2007) etc. to identify Clinical Trials of ImmunoCard STAT HpSA for the primary diagnosis of H. pylori infection. Meta-analysis was conducted using the method recommended by The Cochrane Collaboration Center.

RESULTSEleven trials were included with pooled sensitivity, pooled specificity as 0.93 (95% CI: 0.91-0.94), 0.93 (95% CI: 0.90- 0.95), respectively. Pooled positive likelihood ratio and pooled negative likelihood ratio were 12.01 (95% CI: 8.90-16.19), 0.08 (95% CI: 0.07-0.11), respectively with the pooled diagnostic odds ratio as 160.14(95% CI :100.43-255.34). The area under the summary receiver operating characteristic (SROC) was 0.974 +/- 0.005.

CONCLUSIONImmunoCard STAT HpSA appeared to be an accurate non-invasive method for the initial diagnosis of H. pylori infection.

Antigens, Bacterial ; immunology ; Feces ; microbiology ; Helicobacter Infections ; diagnosis ; immunology ; Helicobacter pylori ; immunology ; isolation & purification ; pathogenicity ; Reagent Kits, Diagnostic ; Sensitivity and Specificity

Objective To study the mechanism and protection of ulinastatin on organ functions in patients with severe disease.Methods Sixty patients in the intensive care unit(ICU)from October 2005 to July 2007 were randomly divided into a control group and an ulinastatin treatment group(each 30 cases).The patients in the control group received the conventional therapy,and the cases in the other treatment group accepted ulinastatin and conventional therapy.According to the disease situations,ulinastatin was administered 200-400 kU once,2-4 times a day,sequentially for 5-7 days.On the day of admission and 3, 5,and 7 days after admission in ICU respectively,blood samples were obtained for measuring alanine aminotransferase(ALT),aspartate aminotransferase(AST),creatinine(Cr),blood urea nitrogen(BUN), activated partial thromboplastin time(APTT),fibrinogen(FIB)and oxygenation index(PaO_2/FiO_2); whether breathing machine or hematodialysis was used and the end results were recorded.Results The rate of usage of breathing machine(23.3%),the incidences of hepatosis(3.3%)and renal dysfunction(10.0%) and fatality(3.3%)in ulinastatin treatment group were obviously lower than those of the control group (63.3%,23.3%,46.7%,10.0%,P0.05).Only one patient received bematodialysis in control group.Conclusion Ulinastatin can protect liver,renal and lung functions markedly and lower the incidence of multiple organ dysfunction syndrome and mortality in patients with severe disease.

CD4+ T cells mainly interact with Sphingosine 1-phosphate (S1P) to regulate immune function through Sphingosine 1-phosphate receptor 1 (S1P1). This study was aimed to investigate the effects of recombinant human granulocyte-colony-stimulating factor (rhG-CSF) mobilization on S1P1 expression in CD4+ T cells of donor's peripheral blood. The CD4+T cells of peripheral blood were isolated by magnetic beads from 17 allo-hematopoietic stem cell transplantation (allo-HSCT) donors before and at fourth day of mobilization with rhG-CSF. The S1P1 expression was detected by real time quantitative PCR in the RNA extracted from CD4+ T cells collected before and after rhG-CSF mobilization. The results showed that the expression of S1P1 was found in CD4+ cells before and after rhG-CSF mobilization, but the expression level of SIP1 in CD4+ cells after rhG-CSF mobilization was significantly lower than that before rhG-CSF mobilization (p<0.01). It is concluded that the mobilization with rhG-CSF obviously down-regulates the expression of S1P1 in CD4+ T cells of donor's peripheral blood.
CD4-Positive T-Lymphocytes ; drug effects ; metabolism ; Granulocyte Colony-Stimulating Factor ; pharmacology ; Hematopoietic Stem Cell Mobilization ; methods ; Hematopoietic Stem Cell Transplantation ; Humans ; Receptors, Lysosphingolipid ; metabolism ; Recombinant Proteins

ObjectiveTo evaluate the prognostic value of MTV on 18F-FDG PET/CT in patients with esophageal cancer.MethodsForty-nine patients with esophageal cancer underwent 18 F-FDG PET/CT scan before surgery.The median follow-up time for the patients was 29 months (range,8- 57 months).The prognostic significance of MTV,age,sex,histologic grade,SUVmax of the primary tumor,tumor size measured on PET/CT,T stage,N stage,M stage,American Joint Committee on Cancer (AJCC) stage,number and location of lymph nodes metastases were assessed by Kaplan-Meier analysis and multivariate Cox model.ResultsIn the univariate analysis,AJCC stage (x2 =16.206,hazard ratio (HR) =1.177,P ＜0.001),N stage (x2 =9.536,HR =10.833,P =0.002),T stage (x2 =5.810,HR=2.397,P=0.016),number of lymph nodes metastases (x2 =11.423,HR =1.567,P =0.001 ),and MTV (x2 =3.872,HR =2.433,P =0.049 ) were significant predictors of survival.Multivariate analysis showed that MTV and AJCC stage were independent predictors of survival (x2 =4.525,HR1.170,P =0.033 ;X2=4.875,HR =3.071,P =0.027).Kaplan-Meier survival curves revealed longer survival time of low-MTV group as compared to high-MTV group ( Log-rank,x2 =4.186,P =0.041 ).ConclusionMTV on 18 F-FDG PET/CT may be an independent prognostic factor in patients with esophageal cancer.

Objective To describe and analyze the characteristics and trend of liver cancer,through data gathered from the Cancer Registry System,in China.Methods Annual registration data on cancer incidents and deaths of 2003-2007,from 32 cancer registries were employed.Crude rates of incidence and mortality,and age-standardized rates by the China population (ASRc) and the world population (ASRw) were calculated.Incidence and mortality rates by age,gender,and by area (urban or rural) were analyzed,and comparison was made internationally,based upon data from the GLOBOCAN 2008.Results The total person-years of 2003-2007 observed from the 32 cancer registries were 255 430 909,in which 197 651 428 from the urban areas,and 57 779 481 from the rural areas.A total of 68 146 incident cases with liver cancer were reported,which accounted for 10.03％(ranked second) of all the registered cases with cancers,with the average annual incidence as 26.68per 100 000 (39.42 in males,13.63 in females).ASRc and ASRw were 13.29 per 100 000 and 17.45per 100 000,respectively.Sex ratios for incidence and mortality were 2.89:1 and 2.72:1,respectively.Incidence rates were 23.91 per 100 000 in urban areas,and 36.15 per 100 000 in rural areas.For mortality rates,they were 22.39 per 100 000 m urban areas and 34.05 per 100 000 in rural areas,respectively.Relative greater differences could be seen amongst these registries.The incidence rates were 10.15-33.85 per 100 000 in the urban areas,and 11.83-78.59 per 100 000 in the rural areas.For mortality rates,they were 13.99-28.45 per 100 000 in urban areas,and 11.02-71.99 per 100 000 in rural areas.According to data from the 184 cancer registries through GLOBOCAN 2008,liver cancer incidence in China was ranked the 5th in males,and 6th in females while mortality was ranked 2nd in males,and 5th in females.Conclusion Liver cancer had been the second most leading malignancy,following lung cancer,with annual incidents and death cases around 360 000 and 350 000,respectively and the figures seemed to be increasing.Nationwide monitoring and research programs on liver cancer should be emphasized.

OBJECTIVETo construct an hybrid bioartificial liver supporting system, and observe its effectiveness and safety on patients with acute on chronic liver failure.

METHODSHybrid bioartificial liver supporting system (HBALSS) was constructed using bioreactor with HepG2 cells transfected with human augmenter of liver regeneration (hALR) gene. 12 acute on chronic liver failure patients were divided into 2 groups randomly. The treatment group was treated with the hybrid bioartificial liver support system. The group underwent plasma exchange was used as control.

RESULTSIn the treatment group, four patients recovered, one patient died of hepatic encephalopathy, one patient died of hepatorenal syndrome, one patient recovered, but died of gastrointestnal bleeding after 1 year. In control group, two patients recovered, one patient underwent orthotropic liver transplantation, and three patients died of liver failure.

CONCLUSIONThe hybrid bioartificial liver supporting system with HepG2 cell line was established successfully and have certain safety and effectiveness on acute on chronic liver failure patients.

Adult ; Bioreactors ; End Stage Liver Disease ; therapy ; Female ; Hep G2 Cells ; Humans ; Liver Failure, Acute ; therapy ; Liver, Artificial ; adverse effects ; utilization ; Male ; Middle Aged ; Treatment Outcome