Objective To construct Cchl1a3 gene R528H knock-in mouse model related to hypokalemic periodic paralysis.Methods ES cells were transfected with Cchl1a3-Konckin targeting vector linearized by Not I digestion , selected in the medium containing both G 418 and ganciclovoir .Resistant clones were screened by PCR and further confirmed by DNA sequencing for correct homologous recombinants .Chimera mice were obtained by routing microinjection of homologous recombined ES cells into blastocysts .Heterozygous mice were obtained by mating .Through heterozygous mice with FLP mice mating , removal of neo gene heterozygous mice were established and identified with the PCR and DNA sequencing . After mating, homozygous offspring were constructed and observed .Results ES cells were successfully transfected withtargeting vector .It was confirmed that 9 resistant clones happened right homologous recombination by PCR and DNA sequencing .7 chimera mice were obtained by microinjection .After breeding the chimeric mice , heterozygous mice were mated FLP mice to obtain 9 heterozygous mice removal of neo gene, the finally obtained 15 homozygous mice with Flp-deleted neo gene.In the developmental stage of sexual maturity , the spirit of the mice, restaurants and activities in good condition, but the gradual emergence of hair removal at 4 months of age, skin ulceration and even death .Conclusions We successfully constructed Cchl1a3 gene R528H mutation homozygous mice.And it laid a foundation for the study of human CACNA1S gene function and to clarify the molecular mechanism of hypokalemic periodic paralysis .

Objective To detect the expression of T cell immunoglobulin mucin 3 (Tim-3) and its relationship with Th17/Treg cytokine isolated from peripheral blood mononuclear cells (PBMCs) and broncho-alveolar lavage fluids (BALF) cells in the asthmatic murine model and to investigate the role of Tim-3 in the occurrence and development of the asthmatic inflammation. Methods The asthmatic murine model was established by the method of ovalbumin (OVA) injection and inhalation. The mice were randomly divided into 2 groups: the normal control and the asthma group. PBMCs and BALF cells in each group were collected. Expression of Tim-3 mRNA was detected by the real-time PCR assay. Levels of CD4+IL-17+ and CD4+CD25+FoxP3+, reflecting expressions of Th17 and Treg respectively , were detected by flow cytometry. The correlationbetween Tim-3 and Th17/Treg level was analyzed. Results Contrast with the normal control group, the expression of Tim-3 mRNA in PBMCs and BALF cells in the asthma group increased significantly (P < 0.01). OVA sensitization and challenges resulted in the increased CD4+IL-17 production and the decreased CD4+CD25+Foxp3+in PBMCs and BALF cells compared with those in the normal control group (both P < 0.05). The ratio of Th17/Treg significantly increased in asthma group (P < 0.05). Tim-3 mRNA expression in PBMCs and BALF cells was positively correlated with CD4+IL-17+ level and Th17/Treg ratio , respectively , and was negatively correlated with CD4+CD25+Foxp3+ level. Conclusions Tim-3 mRNA level in PBMCs and BALF cells in the asthma group was increased , indicating that Tim-3 might take part in the occurrence and development of asthmatic inflammation and has a very close relationship with the Th17/Treg imbalance.

Objective To investigate the serum level of NT-pro-BNP in patients with acute ischemic stroke and to determine whether NT-pro-BNP levels were associated with the death within 15 days of stroke onset. Methods Two hundard twenty-six consecutive patients with acute ischemic stroke within 48 hours of onset were enrolled in this study. We measured plasma NT-pro-BNP within 72 h and recorded the NIHSS score on admission. Patients were divided into two groups: the deceased group, who died within 15 days, and the survival group. The factors associated with the death within 15 d of stroke onset were investigated by using multivariate logistic regression analysis. Results Twenty-four (10.6%) patients died with 15 days of stroke onset. The incidence of atrial fibrillation, cardioembolism and large infarc?tion, the mean ± SD of NIHSS score, age, glucose level and creatinine were significantly higher in the deceased group than in the survival group (P<0.001). On the other hand, the mean ± SD of LVEF, albumin, LDL-C, and total-cholester?ol were significantly lower in the deceased group than in the survival group(P<0.05 ). The median of the plasma NT-pro-BNP level was significantly higher in the deceased group than in the survival group (2598.5 vs. 190.4 pg/mL, P<0.001). The optimal cut-off level, sensitivity, specificity and ROC area of NT-pro-BNP levels to distinguish the de?ceased group from the survival group were 955.2 pg/mL, 83.3%and 82.2%, 0.906, respectively. Binary logistic regression analysis demonstrated that NIHSS score of ≥13 (OR=56.18, 95% CI=9.06 to 348.40, P =0.000) , plasma Lg NT-Pro-BNP level (OR=38.79, 95%CI=6.52 to 230.95, P=0.000) , and the size of infarction (OR=8.73, 95%CI=1.11~68.88, P=0.040) were independent factors associated with the death within acute phase of stroke. Conclusions The plas?ma NT-pro-BNP level can predict the death of stroke patients within 15 days of stroke onset.

Objective:To study the total serum IgE level of infant's asthmatic diseases in acute attacks and its effect on the development of wheezing.Method:The serum IgE levels of 964 children(younger than two years old)diag- nosed as asthmatic bronchitis or bronehiolitis were measured by immune dispersion nephelometer.Result:78.9% childrens had high E levels(females took up 25.9% and males,74.1%),asthmatic bronchitis accounted for 92.3% of the total and bronehiolitis,60.3%.Of the 328 infants involved in our research,76 were diagnosed as asthma at last,which accoun- ted for 23.2%.Conclusion:78.9% infants had a high level of IgE in an acute phase.There was a prominent relativity be- tween IgE and the development of asthma.The increased IgE level of asthmatic bronchitis was obviously higher than that of the bronchiolitis.There appeared to be a connection between the IgE level and the occurrence of asthmatic bronchitis to- gether with the development of asthma.

Objective To investigate the changes and clinical significance of the levels of matrix metallopro-teinase -9(MMP -9)in the serum and bronchoalveolar lavage fluid (BALF)in children with asthma whose had differ-ent levels of 25 -hydroxyl -vitamin D3 [25 (OH)D3 ]in the serum.Methods Fifty children with asthma between January 201 3 and January 201 5 were enrolled as the asthma group,based on the disease severity,and the patients were divided into the moderate to severe group (37 cases)and the mild group (1 3 cases),while 20 children with abnormal airway or tracheal foreign body were as a control group.The levels of 25 (OH)D3 and MMP -9 in the serum,levels of MMP -9 in BALF were measured and compared by using enzyme linked immunosorbent assay (ELISA)me-thod.Re-sults The level of serum 25 (OH)D3 in the asthma group[(1 9.5 ±6.4)μg/L]was lower than that in the control group[(39.3 ±7.1 )μg/L ],and there was a statistical difference between 2 groups(P <0.05).The level of serum 25(OH)D3 in moderate to severe group[(1 6.6 ±4.1 )μg/L]was lower than that in the mild group[(27.9 ±4.5)μg/L],there were statistically significant difference between 2 groups(P <0.05).There was no significant difference in the total number of cells and the percentage of macrophages among different serum 25(OH)D3 levels of children with asthma and the control group(P >0.05).The percentages of neutrophils,eosinophils and epithelial cells in BALF were significantly higher in different serum 25(OH)D3 levels of children with asthma than those of the control group(all P <0.05).There were significant difference levels of MMP -9 in the serum and BALF among different serum 25(OH)D3 levels of children with asthma and the control group(all P <0.05).In children with asthma,the levels of 25(OH)D3 in serum were significantly negatively correlated with the percentages of neutrophils,eosinophils and epithelial cells in BALF (r =-0.683,-0.795,-0.670,all P <0.05 ),exiting a significantly negatively correlation also seen between the serum levels of 25(OH)D3 and MMP -9 (r =-0.796,P <0.05).Conclusions Children with asthma often have low levels of serum 25 -(OH)D3 ,25(OH)D3 and MMP -9 may be involved in airway inflammation and airway remo-deling in children with asthma,and they may involve in the occurrence and development of asthma.

Previous studies have shown that expression of the interferon-sensitive gene (ISG)I5 protease UBP43 is increased in the liver biopsy specimens of patients who do not respond to interferon (IFN)-α therapy. We hypothesized that UBP43 might hinder the ability of IFN to inhibit HBV replication. In this study, we investigated whether vector-based siRNA promoted by Hi (psiUBP43) could enhance IFN inhibiting HBV replication in cell culture. UBP43 was specifically silenced using shRNA. In HepG2.2.15 cells, the HBeAg and HBV DNA levels were significantly reduced by IFN after transfection of shRNA, imphicated that vector-based siRNA promoted by HI (psiUBP43) could enhance IFN inhibiting HBV replication in cell culture. These data suggest that UBP43 modulates the anti-HBV type I IFN response, and is a possible therapeutic target for the treatment of HBV infection.

Objective To investigate the effect of PARP inhibitor 3?aminobenzamide(3?AB)on cortical neurons in streptozotocin?induced diabe?tes mellitus(DM)rats and the mechanism. Methods A total of 60 rats were divided into 3 groups randomly:sham?operated control group,DM group and DM+3?AB group,20 rats in each group. Morris maze was used to detect learning and memory abilities in each group. Spectrophotometer assay was used to detect the levels of superoxide dismutase(SOD),glutathione peroxidase(GSH?Px)and malondialdehyde(MDA)in cortex. West?ern blot was used to determine the expression of poly(ADP?ribose)polymerase 1(PARP1)in cortex. Immunohistochemical staining was used to de?termine the expression of caspase?3?immunoreactive neurons in cortex. Results DM rats showed significantly declined learning and memory abili?ties. Compared with the control group,the levels of SOD and GSH?Px were significantly reduced(P<0.01)and the level of MDA was significantly up?regulated(P<0.01)in the DM group. Compared with the DM group,the levels of SOD and GSH?Px were significantly higher(P<0.05)and the level of MDA was significantly lower in the DM+3?AB group(P<0.05). The expression level of PARP1 was significantly up?regulated in the DM group and was significantly decreased in the DM+3?AB group(P<0.01). The level of caspase?3 was significantly higher in the DM group than in the control group(P<0.01),and was significantly decreased in the DM+3?AB group(P<0.01). Conclusion 3?AB protected cortical neurons from apoptosis in DM rats by inhibition of PARP1 and alleviation of oxidative stress.

Objective To investigate the clinical value of arterial first approach in laparoscopic pancreaticoduodenectomy (LPD).Methods The retrospective cohort study was conducted.The clinicopathological data of 181 patients with pancreatic head and periampullay tumors who underwent LPD in the Affiliated Tongji Hospital of Huazhong University of Science and Technology between October 2014 and December 2016 were collected.Among 181 patients,96 using arterial first approach and 85 using traditional approach were respectively allocated into the experimental group and the control group.Surgery was applied to patients in the same doctors' team,and there were the same extent of surgical resection,range of lymph node dissection and digestive tract reconstruction.Observation indicators:(1) intraoperative situation;(2) postoperative situation;(3) followup and survival situations.Follow-up using outpatient examination and telephone interview was performed to detect the tumor-free survival up to February 2017.Measurement data with normal distribution were represented as x±s,and comparison between groups was analyzed using the t test.Measurement data with skewed distribution were described as M (range).Comparison of count data were analyzed using the chi-square test or Fisher exact probability.Results (1) Intraoperative situation:all the patients underwent successful LPD.Overall operation time and time of digestive tract reconstruction were respectively (268 ± 20) minutes,(33 ± 10) minutes in the experimental group and (285±25)minutes,(30± 17)minutes in the control group,with no statistically significant difference between 2 groups (t =8.529,2.741,P＞ 0.05).Time of tumor resection with superior mesenteric venous invasion were respectively (216± 13)minutes and (264±22)minutes in the experimental and control groups,with a statistically significant difference between the 2 groups (t=41.826,P＜0.05).Time of tumor resection without superior mesenteric venous invasion were respectively (224± 14) minutes and (215±21) minutes in the experimental and control groups,with no statistically significant difference between the 2 groups (t =7.423,P＞ 0.05).Volumes of intraoperative blood loss and blood transfusion were respectively (99± 16)mL,(1.3±0.8)U in the experimental group and (131±27)mL,(2.8±1.2)U in the control group,with statistically significant differences between the 2 groups (t =3.670,0.562,P＜ 0.05).Five and 8 patients had intraoperative blood transfusion in the experimental and control groups,showing no statistically significant difference between the 2 groups (x2=1.195,P＞0.05).(2) Postoperative situation:time of drainage tube removal and duration of hospital stay were respectively (5.8±2.4)days,(18.3±6.3) days in the experimental group and (6.3±3.6)days,(19.6±7.1) days in the control group,with no statistically significant difference between the 2 groups (t =0.498,1.305,P＞0.05).Eleven patients in the experimental group had postoperative early complications,including 8with grade A pancreatic fistula (4 combined with diarrhea,2 combined with biliary fistula,1 combined with delayed gastric emptying and 1 with single pancreatic fistula),3 with grade B pancreatic fistula (2 combined with intra-abdominal hemorrhage and 1 combined with intra-abdominal infection).One patient with intra-abdominal hemorrhage in the experimental group died after treatment failure.Twelve patients in the control group had postoperative early complications,including 6 with grade A pancreatic fistula (2 combined with biliary fistula,2 combined with delayed gastric emptying,1 combined with diarrhea,1 combined with digestive tract hemorrhage),3 with grade B pancreatic fistula and intra-abdominal hemorrhage (2 combined with infection,including 1 death) and 3 with diarrhea.Other patients with complications were cured by symptomatic and supportive treatment.There was no statistically significant difference in overall complications between the 2 groups (x2 =0.287,P＞0.05).Results of postoperative pathological examination showed that case with R0 resection was 93 and 76 in the experimental and control groups,with a statistically significant difference between the 2 groups (x2 =4.057,P＜0.05).(3) Follow-up and survival situations:179 patients were followed up for 2-28 months,with a median time of 14 months.Postoperative 6-month tumor-free survival rate was 92.7％ (89/96) and 88.2％(75/85) in the experimental and control groups,with no statistically significant difference between the 2 groups (x2=1.060,P＞0.05).Conclusion Arterial first approach in LPD could significantly shorten the time of tumor resection of patients with superior mesenteric artery invading pancreatic head and periampullay region,significantly reduce the volumes of intraoperative blood loss and blood transfusion,and increase the rate of R0 resection.

ObjectiveTo investigate the safety, feasibility, and efficacy of total laparoscopic hepatectomy in the treatment of hepatocellular carcinoma (HCC) in the left lateral lobe. MethodsA case-matched analysis was performed between 25 patients with HCC in the left lateral lobe confirmed by postoperative pathological examination, who were admitted to Panzhihua Central Hospital and underwent total laparoscopic left lobe resection (LLLR) from April 2012 to April 2015, and 25 patients with HCC who underwent open left lobe resection (OLLR) during the same period. The t-test was used for comparison of continuous data between the two groups, and the chi-square test or Fisher′s exact test was used for comparison of catagorical data between the two groups; the Kaplan-Meier method was used for survival analysis, and the log-rank test was used for comparison of survival curves. ResultsThe time of operation, number of patients undergoing intraoperative blood transfusion, and number of patients with R0 resection margin showed no significant differences between the LLLR group and the OLLR group, but intraoperative blood loss (216.4±15.39 ml vs 273.2±16.65 ml), mean hospital stay (6.92±0.29 min vs 10.32±052 min), and postoperative complications (5 cases vs 12 cases) showed significant differences between the two groups (all P＜0.05). The 1- and 3-year overall survival rates and progression-free survival showed no significant differences between the LLLR group and the OLLR group. ConclusionLLLR and OLLR have similar long-term efficacy in the treatment of HCC in the left lateral lobe, and LLLR has advantages in the aspects of intraoperative blood loss, postoperative complications, and length of hospital stay.

Objective To investigate whether myocardial perfusion impairment and left ventricular (LV) longituadinal systolic dysfunction had happened in patients with coronary slow flow(CSF),and to assess the relationship between them by myocardial contrast echocardiography(MCE) and speckle tracking imaging(STI).Methods A total of 55 patients underwent coronary angiography for angina were enrolled,of those 35 with coronary slow flow phenomenon as CSF group,20 patients with normal coronary angiography as control group.STI and MCE were performed from the apical 4-,3 and 2 chamber views at baseline and after low-dose dobutamine stress echocardiography (LDDSE) in CSF group and control group.STI derived LV global longitudinal strain(GLS) and GLS amplitude of variation(△GLS),and MCE derived myocardial blood flow(MBF),and myocardial flow reserve (MFR) were obtained.And the correlation between GLS/△GLS and MBF/MFR was analyzed.Results At baseline,the GLS and MBF were similar between CSF group and control group(P ＞0.05).After LDDSE,both GLS and MBF were significantly increased in two groups (P ＜0.05).The GLS,△GLS,MBF,and MFR in CSF group were significantly lower than those in control group(P ＜0.05).There was no significant correlation between GLS and MBF at baseline in the CSF group(r =-0.274,P =0.111).However,after LDDSE significant correlation existed between GLS and MBF(r =-0.630,P =0.000).Conclusions LV longituadinal systolic function is impaired in patients with CSF under dobutamine stress test,the impairment of MBF and MFR could be an important contributor to the decrease of LV longituadinal systolic function.