Objective To investigate the expressions of Toll-like receptor 4 (TLR4) and high mobility group box 1 (HMGB1) expression on distant tissue during the intestinal ischemia/reperfusion and the effects of ω-3 polyunsaturated fatty acids (ω-3 PUFAs) intervention in rats.Methods Forty-eight Sprague-Dawley male rats,weighing (281.50 ± 22.68) g,were randomly divided into three groups (n =16) after gastrostomy:normal diet (N) group,enteral nutrition (EN) group and EN plus ω-3 PUFAs (PUFA) group.Each group was further divided into lymph drainage (I/R + D) and non-drainage (I/R) sub-groups (n =8 each) according to whether treated with intestinal lymph drainage.All the rats were subjected to 60 min ischemia by clamping the superior mesenteric artery,followed by 120 min reperfusion,while the rats in the I/R + D subgroups were treated with intestinal lymph drainage for 180 min at the same time.Results The interleukin-6 level in lymph in N (I/R + D) group was significantly higher than in the EN (I/R + D) and PUFA (I/R + D) groups (PUFA vs EN vs N:(154.57 ±69.30) ng/L vs (97.58 ±40.34) ng/L vs (85.35 ±23.93) ng/L,P =0.021).Besides,the serum level of HMGB1 in PUFA (I/R + D) group was significantly lower compared to the other 5 groups [PUFA (I/R) vs EN (I/R) vs N (I/R) vs PUFA (I/R + D) vs EN (I/R + D) vs N (I/R + D):(2.95 ± 1.17) μg/L vs (3.86 ±0.99) μg/L vs (4.45 ± 1.73) μg/L vs (1.71 ±1.41) μg/Lvs (2.11±0.56) μg/Lvs (3.13 ±0.79) μg/L,P=0.000],and it also decreased in the PUFA (I/R) and EN (I/R) groups than the N (I/R) group (respectively,P ＜ 0.05).Furthermore,the serum endotoxin level in PUFA (I/R) group was significantly lower compared to the N (I/R) and EN (I/ R) groups[PUFA(I/R) vsPUFA (I/R+D) vsEN (I/R) vs N (I/R):(0.020±0.004) EU/mlvs (0.028 ±0.006) EU/ml vs (0.028 ±0.005) EU/ml vs (0.018 ±0.006) EU/ml,P=0.014].Together the serum tumor necrosis factor-α level in both PUFA (I/R) and PUFA (I/R + D) groups were significantly lower than theEN (I/R),N (I/R) and N (I/R+D) groups [PUFA (I/R+D) vs PUFA (I/R) vs EN (I/R) vsN (I/R) vs N (I/R+D):(12.03 ±6.57) ng/L vs (14.32 ±6.11) ng/Lvs (23.27 ±15.60)ng/L vs (27.42 ± 10.37) ng/L vs (26.87 ± 5.30) ng/L,P =0.013].The jejunum and ileum mucosa in all the I/R groups showed swelling and atrophy and appeared fragile,while the PUFA groups showed less yellow staining and injury than the other two groups (P ＜ 0.05,respectively).In addition,the expressions of TLR4 mRNA in jejunum,ileum,and liver in all the drainage groups were respectively lower than the corresponding non-drainage groups [jejunum:PUFA (I/R) vs EN (I/R) vs N (I/R) vs PUFA (I/R+D) vs EN (I/R+D) vsN (I/R+D):2.32±0.62vs3.08±1.29vs3.50±2.44vs 1.62±0.79vs 1.67±1.11 vs 1.94±0.81,P=0.025; ileum:PUFA (1/R) vsEN (1/R) vsN (1/R) vs PUFA (1/R+D) vsEN (1/R+D) vs N (1/R+D):2.67±1.08 vs 5.22 ± 3.96 vs 6.95 ±4.92 vs 1.70±0.68 vs 1.80±0.29 vs3.68±1.47,P=0.012; liver:PUFA (1/R)vsEN (1/R)vsN (1/R)vs PUFA (1/R+D)vsEN (1/R+D)vsN (1/R+D):5.67 ±1.94 vs 7.50 ±3.89 vs 7.18 ±4.55 vs 1.70 ±0.86 vs 3.90 ± 1.95 vs 4.12 ±2.11,P =0.001],which was consistent with the reduction of HMGB1 and the decrease of nuclear factor-κB activity in intestine,liver,and lung (P =0.000).Conclusions Lymph drainage and ω-3 PUFAs intervention can reduce the production of HMGB1 and inflammation factors,inhibit the expression of HMGB1 and TLR4 mRNA,and thus alleviate distant tissue injury caused by intestinal L/R.

Objective To evaluate the safety and efficacy of ultrosonography-guided two-stage ERCP for management of emergent pancreatobiliary diseases during pregnancy.Methods The clinical data of 32 consecutive pregnant women with suspected emergent pancreatobiliary diseases,who were treated with two-stage ultrasonography-guided ERCP from January 2005 to May 2010,were analyzed retrospectively.All patients were treated with ERCP in two stages. In stage Ⅰ,all the patients underwent ultrasonographyguided biliary cannulation,sphincterotomy with small incision and plastic stent placement during pregnancy.In stage Ⅱ,routine ERCP was performed in two weeks after delivery.The success rate of first ERCP,relief of clinical manifestation,improvement of lab indices,maternal and fetal complications were evaluated.Results All patients presented with fever,biliary colic and jaundice.Abdominal ultrasonography or magnetic resonance cholagiopancreatography (MRCP) revealed dilated common bile duct (CBD) in all patients,with CBD stones in 24 and acute cholangitis in 8.Bile duct cannulation and stent placement were successful in all patients.All patients markedly improved after first ERCP,with laboratory indices significantly improved at 1week after the procedure ( P ＜ 0.01 ).In stage Ⅱ,CBD stones were retrieved in 24 patients under ERCP,including 2 cases of mechanical lithotripsy.Bile duct stenosis was found in 3 patients and plastic stents were replaced.Biliary bleeding occurred in 1 case,mild acute pancreatitis in another,with a complication rate at 6.25％.Patients and babies were followed up for 12 months and found to be healthy.Conclusion Twostage ultrosonography-guided ERCP is safe and effective for pregnant patients with emergent pancreatobiliary diseases,which might be regarded as the first choice for such patients to avoid radiation.

Objective To investigate and analyze the situation of urban and rural neglected children aged 3-6,in China,so as to provide basis for the analysis and comparison on relevant risk factors.Methods 1163 urban children aged 3-6 (with 49.6％ males and 4.5％ with minority ethnicity) were investigated from 25 cities of 14 provinces,autonomous regions and municipalities in the whole country.Multi-stage stratified cluster sampling method was used.Again,using the same sampling method,4096 rural children (of whom 50.6％ were males with 6.2％ as minorities) were chosen from 26 cities of 10 provinces or municipalities.Identification of children being neglected was based on “Child Neglect Evaluation Norms of Children Aged 3-6 Years in Urban/Rural China”.SPSS-Windows 13.0 was employed for data analysis.Scores,frequency/degrees,age,sex and types (physical,emotional,educational,safety,medical and social) of children under negligence on every group of the regions,were calculated.x2 test (Chi-Square) and Analysis of variance (ANOVA) were processed to determine the significance of their differences.Results The overall frequencies of negligence were 28.0％ and 53.7％ respectively among the urban and rural children aged 3-6,while the total degrees of negligence were 42.2 and 44.4 respectively.Significant difference was found between children from the urban and the rural areas (P＜0.05).Significant difference was also found between urban and rural children on every age group (P＜0.05).The frequencies of negligence among males were 32.6％ and 55.9％ respectively in urban and rural areas while among females,the figures appeared to be 23.7％ and 51.6％ respectively.The degrees of negligence were 42.7 and 44.6 among male while 41.8 and 44.3 among female children,in the urban or rural areas.Significant differences were found on male or female between urban and rural groups (P＜0.05).Frequencies of negligence in urban children aged 3-6 for the six types were from 5.1％ to 12.9％,with the frequency in rural areas as 13.1％-26.6％.Significant difference was found between urban and rural group for any other type (P＜0.05),in addition to the safety type.The degrees of negligence in urban children aged 3-6 for the different type were between 39.4 and 43.4,while in the rural areas as from 36.5 to 48.2,with significant difference for every type (P＜0.05).The degrees of negligence related to education,emotion,or physical strength were more serious on children from the urban than from the rural areas.The highest frequency of child negligence was seen in the single-parent families on both urban and rural groups (42.9％ and 60.0％ respectively),with no significant difference found (P＞0.05).The urban and rural children aged 3-6 were mainly involved in single item of negligence,with incidence rates as 16.5％ and 22.7％ and proportions as 58.9％ and 45.1％ respectively,despite the factors as age or sex.Conclusion There were large differences on the situation of negligence between the urban and rural children aged 3-6.The frequencies and degrees of negligence in every age group and different sex for children living in the rural areas were higher than those urban children.The frequency of negligence among boys was higher than girls for both urban and rural areas.The rural children had suffered more serious negligence than the urban children at any other type,in addition to the ‘ safety'.Both urban and rural children had the highest frequency of negligence in single-parent family,and were mainly suffered from single item of negligence.

Objective To study the effect of a government-provided-free highly activeantiretroviral treatment (HAART) program,on the reduction of mortality and relevant risk factorsamong adult (AIDS) patients in Henan province.Methods Data on the survival and deaths of AIDSpatients were collected from the National HAART reporting system between 2003 and 2009.Coxproportional hazards model was applied to analyze those factors that affecting the survival time of thepatients.Results 24 669 cases were enrolled to this study in Henan province,from 2003 to 2009.The overall mortality declined from 20.1/100 person-years in 2003 to 5.1/100 person-years in 2009.There was significant difference between the survival curves of different CD4 + T counts and differentnumbers of opportunistic infection syndromes.Results from the Multivariate Cox proportional hazardsregression analysis indicated that CD4+T cell counts ( ＞200 cells/μl,50-199 cells/μ l) was a riskfactor to death,with hazard ratio as 4.4 and 2.0 respectively.Hb of the patients that lower than 90,was a risk factor to death with the HR=1.8.Number of opportunistic infection (OIs) episodes was arisk factor to the mortality (HR=1.7).In addition,other risk factors would include age ( ≥60 years,old),being male,unmarried or divorced,ALT＞100 μl,and other routes of infection,other thanFormer Plasma Donors (FPDs),with HR as 2.2,1.6,1.5,1.3 and 1.2.However,the protectivefactors would include:(1)the earlier the HAART began,the longer the survival time would last(HR =0.8 ) ; (2) when one spouse had already had the infection of HIV,it seemed helpful for the otherspouse to live longer (HR=0.8).Conclusion The National Free Treatment Program hadsignificantly reduced the AIDS mortality rate.Some effective measures should be further taken tomonitor the CD4 + T and the opportunistic infection of the AIDS patients.Patients who were in need totake the HARRT should be adopted into the ART timely,At the same time,the occurrence ofopportunistic infections should be actively prevented.

Objective To evaluate clinical application of phage amplified biologically assay (PhaB) in susceptibility test of Mycobacterium tuberculosis (MTB) in sputum. Methods The drug susceptibility of MTB was detected by PhaB in 143 patients with sputum-positive pulmonary tuberculosis (PTB),and the chemotherapy regimens were adjusted according to the results of susceptibility test.Independent samples t-tests were used for comparison of means.Count numbers were compared with Chisquare test.If there were count number of 0,Fisher probabilities should be used.ResultsThe total positive rate of PhaB was 94.4％ (135/143) with no differences between three types of PTB (x2 =1.886,P ＞ 0.05 ).The duration of testing for PhaB group was (6.6 ± 1.8) days,while for control was (29.4 ±8.7) days (t =29.01,P ＜ 0.01 ).Compared with control group,the 2-month negative-conversion rate (63.2％ vs.35.1％,x2 =3.989,P ＜ 0.05 ) and cure rate ( 100％ vs.78.4％,P ＜ 0.05 ) of PhaB group in type Ⅱ patients were significantly higher.But there were no differences between PhaB and control groups in type Ⅰ and Ⅲ PTB patients.ConclusionThe results of PhaB drug susceptibility test can be helpful for choosing effective chemotherapy regimen for PTB patients rapidly.

Objective To develop a system to rescue virus by intracellular expression of T7 RNA Polymerase. Methods The gene of T7 RNA Polymerase was amplified and cloned to VR1012 by molecular biological technology. The expression plasmid VR-1a was then identified. VR-1a and EV71 infectious plasmid were co-transfected in Veto cell. CPE was observed and viral gene viral antigen were detected.Results The gene of T7 RNA Polymerase was successfully cloned into vector VR1012. Vere cell developed to CPE after being transfected VR-1a and EV71 infectious plasmid. EV71 gene was amplified by RT-PCR from the culture. EV71 antigen was also detected by ELISA. Conclusion The method can be used to rescue virus. It could apply to immunologic research of EV71 DNA vaccine.

Objective To explore the influence of nonylphenol(NP) on the filial generation rats spatial learning and memory capacity which were exposed to the parent rat during its pregnancy. Methods At the first day of the pregnancy, the rats were divided into four groups, which were orally administered with NP at doses of 0,50, 100 and 200 mg/kg,respectively, on gestational days 9～15. Cognitive function was tested by Morris water maze and step-down test. The ultrastructure of hippocampus tissues were observed by electronic microscope.Result The escape latency extended ((61.14±5.92) s) and erroring time increased ((4.57±1.13)times) in Morris water maze, and step down latency extended ((37.5±6.3)s), step through latency shortened((97.8±11.0)s) and erroring time increased ((3.0±1.4) times) in step down test in the N P 200 mg/kg treated groups (P<0 05). The correspond indexes were separately (35.85±4. 29) s, (2.57±0.97) time, (27.1±3.8) s,(172.0±89.2)s and(0.9±0.7)time in control group. Compared to the control group, there were significant differences in the results of the water maze test and step-down test between NP 200 mg/kg and the control group (P <005). The changes of the uhrastructure were found among the hippocampus neurons of NP 200 mg/kg group that characterized with chromatin condensed,clumped in circa-nuclei and mitochondrial tumefaction and vacuolization.Conclusion Exposures to NP during gestation might decreased abilities of spatial learning and memory capacity on F1 rats significantly.

Objective To rational design HBV therapeutic vaccine candidate and evaluate their specific immunity to HBV in mice.Methods Based on our previous data of HBV protein vaccine consisting of S-PreS1 fusion particle.We first construct a novel DNA vaccine candidate,pVRC-HBSS1,which consisting of S (an:1-223) and PreS1 (an:21-47) fuse gene,then confirm the expression of the DNA vaccine by Western blotting,and followed by vaccination using prime boost strategy,ie,Intradermal injection of DNA vaccine with gene electroporation (EP) in BALB/c mice after twice injection of different HBSS1 protein vaccines (combination with different adjuvants).The immune response was measured by ELISA and IFN-gamma ELISPOT.Result The novel DNA vaccine candidate could effectively express in vitro,boost with single intradermal injection of HBV DNA vaccine via EP can significantly enhance the surface antigen (S)-specific cellular immune responses (IFN-γ,ELISpot analysis) and PreS1-specific antibody levels,especially in the group primed with protein vaccine in combination with alum adjuvant.Conclusion Boost with the novel HBV DNA vaccine followed prime with HBV protein vaccine could induced a higher anti-HBV T cell response in mice than vaccination with the HBSS1 particle-like protein vaccine only.This prime-boost vaccination may serve as a promising way to develop and optimize the novel HBV therapeutic vaccine.

Objective To evaluate the effect of protecting parathyroid glands in situ in the operation of total thyroidectomy by detecting parathyroid hormone after the operation.Methods In the surgical team, 1019 consecutive patients with thyroid diseases were treated with total thyroidectomy.During the operation, parathyroid glands were protected in situ with correctly identifying the parathyroid glands, precisely dissecting its envelope and protecting its blood supply.Serum calcium level and parathyroid hormone were measured before and 24 hours after operation. The patients who had symptomatic hypocalcemia or hypoparathyroidism were given supportive treatment and followed-up. Results At least one of the parathyroid glands was preserved and remained in situ in all cases.Eighty-nine cases (8.7%) had decreased parathyroid hormone levels and 42 cases (4.1%) had complicated symptomatic hypocalcemia.The symptoms of hypocalcemia in all these cases could be controlled by supportive treatment, and serum calcium level and parathyroid hormone had all recovered 1 - 6 months later.If 3 and 4 parathyroid were conserved in situ, the postoperative complication rate was significantly lower than those with 1 and 2 parathyroid conserved(decreased PTH 69/999 vs 20/20, symptoms of hypocalcemia 25/999 vs 17/20, all P <0.01).Conclusion The techniques to protect parathyroid glands in situ are effective measure to prevent the postoperative hypoparathyroidism in total thyroidectomy.

Objective The aim of this study is to examine the expressions of Toll like receptor (TLR) 7 and TLR9 in the peripheral blood B lymphocytes of SLE patients and to analyze the correlation between TLR7/9 and clinical parameters. Methods lntracellular expression of TLR7/9 in the peripheral blood CD19+Blymphocytes was analyzed in 50 SLE patients and 30 healthy controls by flow cytometry. The difference of intracellular TLR7/9 expression levels in two groups was compared. Furthermore,the correlation between TLR7/9 expression and clinical parameters such as ESR, CRP, complement 3 (C3), complement 4 (CA), the level of serum IgG, anti-double stranded DNA antibody, anti-nuclear antibodies, SLEDAI score and urine protein excretion level, were analyzed. Results Compared with healthy subjects, the proportion of B cells expressing TLR7 and TLR9 was higher among SLE patients. Positive correlation was observed between TLR7 expression levels and clinical measurement of the SLEDAI and ESR. Negative correlation was observed between TLR7 expression levels and serum C3 levels. Positive correlation was observed between TLR9 expression levels and SLEDAI scores. Negative correlation was observed between TLR9 expression levels and serum C3 levels. Conclusion TLR7 and TLR9 expression is increased in the peripheral blood B cells of SLE patients, and correlates well with clinical parameters.