The main impacting factors on implementing clinical pathway were analyzed by applying fishbone diagram.The factors include policy factors,organizational factors and personal factors.It was suggested to improve the trial work of implementing clinical pathway management by reforming the medical care payment system,putting the clinical pathway management into the evaluation system,building up medical information system,putting more efforts on promotion and enhancing cooperation among related departments.

Objective To explore the method of primary isolation, cultivation and identification of rat brain microvessel pericytes. Methods The brain tissue of 10 3 week-old Wistar rats was separated sterilely. The brain microvessel fragments were separated using two-step enzyme digestion and one-step gradient centrifugation and were seeded in 35-mm dishes for primary culture. The cell morphology was observed by phase contrast microscopy; the immunofluorescence assay was used to identify the associated antigns, such as the α-smooth muscle actin (α-SMA), neuron-glial antigen 2 (NG2), von Willebrand factor (vWF), and glial fibrillary acidic protein (GFAP). Methyl thiazolyl tetrazolium was used to determine the cell growth curve. Results Pericytes climbed out from the adherent brain microvascular fragments around,showing polygonal, and the cell fusion was 95％ after 12-14 days. Immunofluorescence staining revealed that the molecular markers of the pericytes α-SMA and NG2 related antigens showed double positive, while the vWF and GFAP related antigens showed double negative and the cultured cells were confirmed as brain microvascular pericytes. The growth rate of primary cells was slower. The passage cells entered into logarithmic growth phase after 36 to 60 hours and entered into plateau phase after 72 to 108 hours. Conclusions This method may successfully isolate rat brain microvascular pericytes with higher purity.

Objective To establish a stable in vitro model of blood-brain barrier (BBB) simulating in vivo state using the primary-cultured rat brain microvascular endothelial cells (BMVECs) and pericytes.Methods The primary rat BMVECs and pericytes were isolated,purified and cultured.The isolated cells were identified by immunocytochemical staining method.An in vitro model of BBB was constructed using Transwell inserts (pore size 0.4 μm) coculture.Its barrier function was evaluated by the 4-hour leakage test,tight junction protein identification,transendothelial resistance detection,and permeability test.The difference between the cocultured model and simple BMVEC model across the membrane resistance values,and the permeability difference of the small molecule sodium fluorescein (Na-F) were compared.Results Confluent BMVEC monolayers demonstrated a typical cobblestone appearance and the pericytes displayed irregular shape and overlapping growth.Immunodouble labeling technique identification showed that the pericytes positively expressed α-srmooth muscle actin (α-SMA) and neuron-glial antigen 2 (NG2); after the fusion of cocultured model endothelial cells,the surface leakage test became positive; immnocytochemical staining shows that a continuous and dense tight junction formed between the endothelial cells; compared to the BMVEC model,the transendothelial electrical resistance of the cocultured model increased significantly (190.762 ± 10.326 Ω/cm2 vs.96.503 ± 8.012 Ω/cm2; t=- 24.489,P ＜0.01),and the permeability decreased significantly (56.149％ ± 3.572％ of the single endothelial model; t =19.330,P ＜ 0.01 ).Conclusions The primary isolated rat BMVECs and pericytes cocultured the morphology,structure and barrier function of in vitro model have the basic characteristics of BBB,and they have provided a useful tool for the research of BBB.

Aim To observe the ameliorating effects of sophocarpine on mice with neuropathic pain induced by spared nerve injury(SNI), and to explore the analgesic mechanism.Methods Sixty male ICR mice were randomly divided into six groups: sham-operated, neuropathic-pain model(SNI), SNI+Pregabalin(Pre) 10 mg·kg-1, SNI+SC 40 mg·kg-1, 20 mg·kg-1 and 10 mg·kg-1 group.Neuropathic-pain mouse models were established by SNI of the sciatic nerve.The paw withdrawal mechanical threshold(PWMT), paw withdrawal thermal latency(PWTL), and cold withdrawal times(CWT) were detected to assess the analgesic effects of sophocarpine on mice with neuropathic pain induced by spared nerve injury.The expressions levels of TLR4, p38 MAPK, IL-1β and TNF-α mRNA and protein in spinal cord tissue were detected by reverse transcription-polymerase chain reaction(RT-PCR) and Western blot methods.Results Compared with sham group, the SNI group′s PWMT decreased, PWTL was shorten, CWT increased, and the expression levels of p38 MAPK, TLR4, TNF-α and IL-1β mRNA and protein in spinal cord tissue were up-regulated.Compared with SNI model group, the sophocarpine 40 mg·kg-1 group′s PWMT increased, PWTL was lengthened, CWT decreased, and the expression levels of p38 MAPK, TLR4, TNF-α and IL-1β mRNA and protein in spinal cord tissue were down-regulated.Conclusion Sophocarpine has the analgesic effects on neuropathic pain induced by SNI, and its mechanism may be related to the down-regulation of TLR4, p38 MAPK, IL-1β and TNF-α expression levels.

Objective To explore the association between X-ray repair cross complementing group 1 (XRCC1)gene rs25487 locus polymorphisms and nonobstructive azoospermia in Hui minority ethic population of Shaanxi Province.Methods We used polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)method for genotyping at XRCC1 gene rs25487 locus in 79 patients with nonobstructive azoospermia and 82 healthy male controls in Hui minority ethic population of Shaanxi Province.Then we analyzed the association be-tween XRCC1 gene rs25487 locus and nonobstructive azoospermia.Results Compared with GG genotype,GA, AA and GA + AA genotypes demonstrated a significantly increased risk for nonobstructive azoospermia (OR =2.286,95% CI 1.1 5 1-4.539;OR =2.202,95% CI 0.753-6.439;OR =2.271,95% CI 1.1 71-4.403),respec-tively.Meanwhile,the A allele frequency was significantly higher in azoospermic patients than in controls (OR =1.582,95% CI 1.005-2.492,P =0.047).Conclusion G→A in XRCC1 gene rs25487 locus is correlated with nonobstructive azoospermia in Hui minority ethic population of Shaanxi province.

Ribavirin is a broad-spectrum antiviral agent and glycyrrhizin has activities of anti-inflammation, immunoregulation and anti-viral infections. To enhance antiviral efficacy and weaken side-effects of ribavirin, antiviral effects of the combination of glycyrrhizin and ribavirin were studied in the present study. Firstly, a mouse model of viral pneumonia was established by inoculation of influenza H1N1 virus. Protective effects of glycyrrhizin and ribavirin used alone or in combination against H1N1 virus infection in mice were evaluated based on the survival rate, lung index and virus titer in lungs of mice. Results showed that the combination of glycyrrhizin and ribavirin significantly inhibited the lung consolidation with a 36% inhibition ratio on the lung swell of infected mice. The combination of the two drugs exhibited synergetic effects on survival of infected mice. The combination of 50 mg · kg(-1) · d(-1) glycyrrhizin and 40 mg · kg(-1) · d(-1) ribavirin resulted a 100% protection for infected mice with a synergetic value of 36, which was significantly higher than the control group and each drug alone. This combination also resulted a significant drop of lung virus titer (P < 0.01), as well as inhibition on the production of proinflammatory cytokines IL-6 (P < 0.01), TNF-α (P < 0.01) and IL-1β (P < 0.05) induced by virus infection compared to the control. The treatment of ribavirin plus glycyrrhizin was more effective in influenza A infection in mice than either compound used alone, which suggested a potential clinical value of the combination of the two agents.

Objective To evaluate the efficacy and safty of antiviral treatment for chronic hepatitis B ( CHB ) patients in second trimester of pregnancy.Methods Seventy-nine CHB patients in second trimester of pregnancy were collected from Hangzhou First People’ s Hospital and Xixi Hospital of Hangzhou during January 2010 to December 2013.Patients were divided into antiviral treatment group ( n=47) and the control group (n=32) according to their own wishes.Patients in antiviral treatment group were given lamivudine or telbivudine treatment plus hepatoprotective medication, while those in control group were only given hepatoprotective medication.All pregnant women were observed for 12 weeks after childbirth and the neonates were followed-up for 6 months after birth.The liver function, HBV DNA loads, HBV serological markers were measured;adverse effects during pregnancy, blocking rates of mother-to-child transmission and the growth of neonates were documented.t test or Chi-square test was used for statistical analysis.Results Alanine aminotransferase ( ALT) normalization rate and HBV DNA negative rate in antiviral treatment group before childbirth were 88.6%(39/44) and 84.1%(37/44) , while those in the control group were 60.0%(18/30) and 0 (χ2 =8.27 and 50.46, P<0.05).After 12 weeks of childbirth, ALT normalization rate and HBV DNA negative rate in antiviral treatment group were both 100.0% (44/44), which were higher than those in control group (90.0%and 0) (χ2 =4.59 and 74.00, P<0.05).HBeAg seroconversion was observed in 1 (2.8%) and 4 (11.1%) patients in antiviral treatment group before and 12 weeks after childbirth, but it was not observed in the control group.The difference in HBeAg seroconversion rate bwteen two groups was not of statistical significance (P>0.05).No patient in antiviral treatment group terminated pregnancy due to abnormal liver function or adverse effect of drugs, while 2 out of 30 patients (6.7%) in the control group terminated the pregnancy, but the difference between two groups was not of statistical significance (χ2 =1.01, P >0.05).Mother-to-child transmission of HBV was successfully blocked in antiviral treatment group, while 3 cases (11.5%) in control group were failed (χ2 =5.19, P<0.05).No significant differences in gestational age, body mass, body length, Apgar score between two groups were observed (t=0.65, 0.84, 0.25 and 0.77, P>0.05).Conclusion Antiviral treatment can improve liver function, inhibit HBV replication and reduce the risk of mother-to-child transmission, and is safe for CHB patients in second trimester of pregnancy.

Objective To evaluate the effect of comprehensive schistosomiasis control measures based on infection source control in plateau mountain areas of Yunnan Province. Methods From 2006 to 2004,four administrative villages were selected as test areas from plateau canyon and plateau basin endemic areas in Jindun Town,Heqing County,two villages each type,and the comprehensive control measures were implemented,including the examination and treatment of schistosomiasis,Oncomela?nia hupensis snail survey and control,health education,improving drinking water and lavatories,banning grazing,constructing sanitary pen of livestock,replacing cattle with machine,etc. The schistosome infection state and snail status in 2006 were treat?ed as the baseline information,and the effect of the comprehensive measures were evaluated. Results The infection rate of hu?man in plateau canyon areas decreased from 4.94%in 2006 to 0.06%in 2014,and that of livestock decreased from 1.11%to 0. In plateau basin areas,there was only 1 case of schistosomiasis found in Xiaolian Village in 2007,and no any other cases found in the other years,the infection rates of livestock dropped from 7.38%to 0. Compared with 2006,the snail areas in the two type areas decreased by 74.89%and 75.30%,respectively,meanwhile,the percentage of snail area,the occurrence rate of frames with snails,as well as the average density of living snails also decreased,and no infected snails were found since 2008. Xidian and Xinzhuang villages in plateau canyon area reached the criteria of schistosomiasis transmission controlled in 2009,and Xiao?lian and Kangfu villages in plateau basin reached the criteria of transmission interrupted in 2014. Conclusions The comprehen?sive schistosomiasis control measures based on infection source control can effectively control the endemic situation of schistoso? miasis in plateau areas of Yunnan Province. In the future,we should pay an equal attention to the infection sources control and snail control to consolidate and amplify the achievement of schistosomiasis control.

Objective To understand the status of water contact and labor practice of residents in plateau mountain schisto?somiasis endemic areas in Yunnan Province so as to provide the evidence for formulating the strategy of prevention and control of schistosomiasis in the next step of work. Methods A total of 412 residents aged 6-65 years old in 2 villages were randomly sampled and investigated with questionnaires for their water contacts and labor practices in plateau valley schistosomiasis endem?ic areas. Then the schistosome infection status of the residents was surveyed with the indirect hemagglutination assay IHA . Re?sults Among the 412 residents investigated the rate of water contact was 88.35% and the main causes of water contact were watering and swimming and playing. The percentages of residents who had schistosomiasis history were 1.16% 0.00%30.80% 3.85% and 0 in them who had swimming and playing water?contacts bathing watering washing hands and others respectively χ2=38.96 P<0.01 . The positives of IHA in the above?mentioned residents were 18.60% 0.00% 37.60%23.08% and 0 respectively χ2=12.61 P<0.05 . Conclusions The productive infested water contact is the main way of schistosome infection. Therefore the changes of labor practices and water contact of the residents are very important for the ef?fective prevention and control of schistosomiasis transmission.

Objective To detect the positive rates of antibodies against avian influenza virus (AIV) subtypes H5, H6, H7 and H9 among people in poultry occupations in Guangdong province and to analyze the transmission of various subtypes of AIV from poultry to human contacts for the prevention and control of novel AIV infection in human beings.Methods Serum specimens were collected from 1066 peo-ple in poultry occupations ( occupational group) and 205 people not in poultry occupations ( non-occupational group) in 10 cities of Guangdong province.The inactivated AIV strains, isolated from poultry or environment of Guangdong province, were used as antigens to detect antibodies against AIV subtypes H5, H6, H7 and H9 by using the hemagglutination inhibition ( HI) assay.Results The positive rates of antibodies against AIV subtypes H5, H6, H7 and H9 carried by people from the occupational group were respectively 0.44%, 0%, 0.30%and 0.30%in 2013 and 1.08%, 0.0%, 0.0%and 0.27%in 2014.Only the anti-H9 anti-bodies were detected in serum samples collected form people in the non-occupational group in 2013 with a positive rate of 0.95%.No significant differences with the positive rates of anti-AIV antibodies were found between the occupational group and the non-occupational group.However, the geometric mean titer ( GMT) of anti-AVI antibodies in people from the occupational group was higher than that of the non-occupational group.Conclusion Although a grand spread of AIV from avian to human is not likely to happen yet, con-tacting with poultry is the risk factor for AIV infection in Guangdong population.A long-term surveillance of anti-AIV antibodies in serum should be strengthened among people in poultry occupations for the timely pre-vention and control of novel AIV outbreak.