Abstract:Objective: To prepare fluorescent nano gene vectors for traceable gene delivery and evaluate the biocompatibility of the nanoparticles. Methods: The fluorescent nanopaticles were constructed by quantum dots (Qdots) encapsulated by chitosan, then characterized by transmission electron microscopy (TEM), fluorospectrophotometer, zeta potential and Fourier transform infrared spectrometry (FTIR). The imaging of tumor-injected xenograft nude mouse was taken to evaluate the in vivo fluorescent signal of CS-Qdots nanoparticles. Gel retardation assays were used to test the DNA binding affinity of CS-Qdots. The expression of green fluorescent protein (GFP) gene delivered in cells by CS-Qdots was observed by confocal laser microscope. The biocompatibility of CS-Qdots was evaluated by cells relative grow rate (RGR) of MTT test, hemolysis rate and acute toxicity tests in mice. Results: The mean particle size of the CS-Qdots nanoparticles was 20-30 nm in the TEM image and the surface charge of them was （28.02 ± 1.15） mV. The characteristic peaks of chitosan were observed on the CS-Qdots FTIR spectra. The light emission peak of CS-Qdots was at 630 nm. Complete retardation was observed for particle-DNA weight ratios over 10∶1, for which the DNA was well packed in the gene-CS-Qdots complexes with a positive surface charge. High level expression of GFP genes delivered into HepG2 cells by CS-Qdots was detected by a confocal laser microscopy. The strong fluorescence signal of CS-Qdots in vivo was observed in xenograft nude mouse imaging. The MTT results indicated that RGRs of cells co-incubated with CS-Qdots at concentrations of 50, 100, 200 and 400 μg/mL were 1.000, 1.000, 0.917, and 0.875, respectively. The RGRs of cells co-incubated with Qdots at the corresponding concentrations were 1.000, 0.850, 0.621 and 0.326. The hemolysis rates of Qdots over the concentration of 100 μg/mL were all above 5%. Oppositely, the hemolysis rates of CS-Qdots were all below 5% at all the concentrations tested. The results of acute toxicity tests in mice at 72 h post injection indicated that compared to the saline-controlled mice, no symptoms of main organ lesions were observed, no abnormality was seen in the blood cells test, and the functions of liver and kidney were normal in the mice. Conclusion: The CS-Qdots fluorescent nanoparticles with high biocompatibility were constructed successfully, which delivered genes into cells efficiently. The nano gene vectors could be traced by fluorescent imaging in vivo and in vitro.

Abstract:Objective：To investigate the resuscitative potential of mycobacteriophage Guo1 on dormant cells of M.tuberculosis.Methods：Dormant cells of M.tuberculosis were induced by potassium-deficient culture of logarithmic phase of M.tuberculosis at 37 ℃，which were proven through drug sensitivity test，transmission electron microscope(TEM) and auramine O-Nile red double fluorescent staining. M.tuberculosis colony count was used to detect whether the dormant cells of M.tuberculosis had been resuscitated by Guo1. Results：Dormant cells of M.tuberculosis were obtained successfully by potassium-deficient cultivation for 30 days at 37 ℃. The concentration of M.tuberculosis in control，supernatant(SN)，TM4 and Guo1 groups was (2.67±0.58)×102，(3.00±1.00)×102，(2.67±0.58)×102，(2.33±0.58)×102 CFU/mL，respectively，after culture for 1 d. And the concentration of M.tuberculosis in above groups increased to(7.33±1.53)×104，(6.67±2.08)×1010，(3.00±1.00)×1010，(3.33±1.53)×1010 CFU/mL at the 15th day，respectively. Conclusion：Mycobacteriophage Guo1 can resuscitate dormant cells of M.tuberculosis.

Abstract:Objective: To investigate the protective effects of formononetin (FN) on the rats with inflammation after focal brain ischemia reperfusion injury. Methods: Sixty adult male SD rats were randomly subjected to either sham surgery, model, FN (8, 16, 32 mg/kg) or nimodipine group (12 mg/kg) for 2 h of brain ischemia and 24 h reperfusion with Longa's score analysis, each with 10. After 24 h of reperfusion, the cerebral infarction volume was determined by TTC staining, and the contents of inflammatory factors in ischemic brain tissue were assayed in each group. In addition, we also examined the changes of nuclear transcription factor-kappaB (NF-κB) and sphingosine kinase 1/sphingosine-1-phosphate (SphK1/S1P) signaling pathway in the ischemic brain tissue. Results: FN each dose group reduced the expression of inflammatory factors, with the inhibition of NF-κB and SphK1/S1P signal pathway significantly. Conclusion: The protective effects of FN on the rats with inflammation after focal brain ischemia reperfusion injury may be related to its inhibition of SphK1/S1P signal pathway.

Abstract:Objective: To study the radiosensitization effects of pemetrexed disodium and gefitinib on human lung adenocarcinoma cells. Methods: Human lung adenocarcinoma A549 cells were treated with pemetrexed and/or gefinitib, and cell proliferation was examined by MTT. Colony formation was performed to detect growth inhibition effects of pemetrexed and/or gefinitib combined with radiation on A549 cells. Flow cytometry was performed to detect the changes of cell cycle and apoptosis. Expressions of intracellular Akt and p-Akt were detected by Western blot. Results: Pemetrexed and gefitinib inhibited cell proliferation of human lung adenocarcinoma A549 cells. Compared with cells treated with radiation only, radiation+pemetrexed, and radiation+gefitinib, the Dq, D0 and SF2 of cells treated with radiation+pemetrexed+gefitinib were significantly decreased. The SERs of cells treated with pemetrexed, gefitinib, and pemetrexed+gefitinib were 1.17, 1.48 and 2.62, respectively. Both pemetrexed and gefitinib induced G1/G0 phase arrest. When combined with radiation, the proportion of S phase cells was further reduced. Both pemetrexed and gefitinib enhanced radiation-induced apoptosis by decreasing p-Akt level. The p-Akt level was lowest in the cells treated with radiation+pemetrexed+gefitinib. Conclusion: Both pemetrexed and gefitinib have radiosensitization effects on A549 cells. Combination of the two drugs significantly improves the inhibitory effect of radiation through reducing S phase cells, enhances radiation induced apoptosis, and inhibits PI3K/Akt signaling pathway. This combination regimen may provide a promising strategy for lung cancer therapy.

Abstract:Objective: To prepare the double acid etching titanium surface modified with calcium-containing nanoflakes and evaluate its effe cts on osteoblast behaviors in vitro. Methods: Through double acid-etching and hydrothermal treatment with a mixed solution of Ca(OH)2/H2O2, two different calcium-containing nanoflakes(the 1 h and the 6 h treatment groups) were synthesized on the double acid etching titanium surfaces. The sandblasting with large grit and acid etching (SLA) titanium surface was used as the control group, and two double acid etching titanium surfaces modified with different calcium-containing nanoflakes were used as two experiment groups. The micro-topography and surface elemental compositions of different titanium surfaces were observed and analyzed. MC3T3-E1 cells were seeded on samples of the three groups to investigate osteoblast behaviors on different titanium surfaces. Results: Two uniform nanoflake-like structures containing trace calcium were prepared on double acid etching titanium surfaces. Compared with the SLA titanium surface, two modified titanium surfaces significantly up-regulated the adhesion, proliferation, ALP activity and expressions of osteogenic-related genes of MC3T3-E1 cells, especially the 1 h treatment group. Conclusion: The double acid etching titanium surface modified with calcium-containing nanoflakes could promote the adhesion, proliferation and differentiation of osteoblasts.

Abstract:Objective: To prepare Ru(bpy)32+(SiO2@Ru) coated silica nanospheres as electrochemiluminescence (electrogenerated chemiluminescence, ECL) biosensor probe, and evaluate its application value in the detection of the hepatitis C core antigen (HCVcAg). Methods: The water-in-oil(W/O) microemulsion method was employed for one-pot synthesis of SiO2@Ru nanoparticles. The as-synthesized SiO2@Ru nanoparticles has a narrow size distribution, which allowed reproducible loading of Ru(bpy)32+ inside the silica shell and HCVcAg, on the silica surface with glutaraldehyde as linkage. The as-synthesized SiO2@Ru could be used as a lable for ultrasensitive detection of HCVcAg through a sandwich immunoassay process. Results: The calibration range of HCVcAg concentration detection was 1-1 000 fmol/L，the lowest detection limit was 3 fmol/L. Compared with clinical RNA HCV, there was a good correlation, linear correlation coefficient was R2=0.937 9. Conclusion: With high sensitivity and good reproducibility, SiO2@Ru could be used as a label in clinical diagnosis, curative effect evaluation and blood screening of blood center.

Abstract:Objective：To investigate gene polymorphsim of human leukocyte antigen G (HLA-G) 3’ untranslated regions (3’-UTR) in Uygur，Xinjiang. Methods：Genomic DNA was extracted from 100 blood samples of Uygur population. Amplification of HLA-G 3’-UTR was performed by PCR. The productions were directly sequenced to analyze the frequency of genotypes and haplotypes.Results：There were 98 samples sequenced successfully.Ten single nucleic polymorphisms were found in HLA-G 3'UTR in Uygur populations consisting of 14bpDel/Ins，+3003C/T，+3010C/G，+3027A/C，+3035C/T，+3092T/G，3121T/C，+3142C/G，+3187A/G and +3196C/G. Strong linkage disequilibrium existed among these SNPs. It showed a higher HLA-G 3’-UTR haplotype diversity including 20 haplotypes and UTR-2 was the most one (24.9%). Conclusion：Polymorphisms and haplotypes of HLA-G 3’-UTR were distinct in Uygur populations. The data provide fundamental and critical information to explore mechanism of HLA-G in the role of genetics，evolution，immunity function and related diseases.

Abstract:Objective: To investigate the application of bootstrap method in the early warning for influenza surveillance with 95%CIs of weekly influenze-like illnesses(ILI)% estimated by bootstrap method specified as the early warning thresholds. Methods: The influenza surveillance data from 2005 to 2014(not including 2006 and 2009) were taken as baseline. Thirty Percent of the highest positive rate of influenza viruses in two years was taken as the gold standard of influenza pandemic. Consistency of the early warning results was compared among bootstrap method, moving average method and the gold standard. We also discussed the application of bootstrap in practice in 2016 based on the surveillance data from 2012 to 2015. Results: Influenza viruses circulated throughout the year and peaked during cold winter and spring season from November to following February and hot summer from June to August. The result of bootstrap method has high consistency with moving average method and gold standard. The bootstrap method was more sensitive in the early warning of influenza pandemic in 2009 compared with moving average method and more consistent with gold standard(kappa=0.728). Conclusion: Bootstrap was a compute-intensive method which was not rely on the distribution of ILI% data, and it can be applied as an alternative and complementary method in the early warning for influenza surveillance.

Abstract:Objective: To investigate the curative effect of minimally invasive hematoma puncture in the treatment of hypertensive cerebral hemorrhage and the effect on the nerve function and the expression of inflammatory factors in patients with hypertensive cerebral hemorrhage. Methods: A total of 74 hypertensive cerebral hemorrhage patients were selected and grouped according to the filing order of hypertensive cerebral hemorrhage from October 2013 to August 2015, 37 cases in each group. The control group was treated with small bone window hematoma, the study group was treated with minimally invasive hematoma puncture. The patients were followed up from 3 to 6 months. After surgery, we compared the statistical operation, clinical efficacy, complications, analysis of preoperative and last follow-up two group of neural function defect score (NIHSS), daily life ability score (BI) and serum inflammatory factor ［tumor necrosis factor alpha (TNF-alpha), interleukin-6 (IL-6), high sensitive C reactive protein (hs-CRP) level change］ of the two groups. Results: ① Operation: The operation time (12.34 ± 5.54) min, intraoperative blood loss (40.31 ± 10.27) mL, ambulation time (5.04 ± 2.82) d, and hospitalization time (16.30 ±2.61) d in the study group were less than those of the control group, the difference was statistically significant (P<0.05); ② Neurological function and ability of daily living: There was no significant difference in NIHSS score and BI score between the former two groups (P>0.05), after operation, the NIHSS score of the study group (4.08±1.35) was lower than that of the control group (6.49±1.13), and the BI score (60.63±8.08) was higher than that of the control group (46.82±6.81), and the difference was statistically significant (P<0.05); ③ Clinical effect: The effective rate of the study group was 91.89% (34/37), which was higher than that of the control group 72.97% (27/37), and the difference was statistically significant (P<0.05); ④ Serum inflammatory factors: There was no significant difference in the levels of hs-CRP, IL-6 and TNF-α in two groups before operation (P>0.05), the levels of serum inflammatory factors in two groups were lower than those before operation, and hs-CRP (7.14± 3.05) mg/L, IL-6 (14.18 ± 3.16) ng/L, TNF-α(40.10±3.03) μg/L of the study group was lower than that of the control group ［(12.11 ± 3.35) mg/L, IL-6 (27.19 ± 3.98) ng/L, TNF-α(53.22 ±3.32) μg/L］, the difference was statistically significant (P<0.05); ⑤ Complication rate: The incidence of complication was 5.41%(2/37) in the study group, which was lower than that in the control group (9/37), and the difference was statistically significant(P<0.05). Conclusion: The effect of minimally invasive hematoma puncture in the treatment of hypertensive cerebral hemorrhage is significant, which can improve neurological function of patients and ability of daily living, reduce the operation time and blood loss during operation, decrease serum levels of inflammatory factors, and improve the treatment effect with high safety.

Abstract:Objective: Through detection of serum cytokines and immune factors from the hospitalized elderly patients with depression, and further analysis of the correlations between concentration variations and clinical symptoms and therapeutic effect, we sought to provide potential objective criteria for diagnosis, evaluation of therapeutic effect and treatment of depression in elderly patients. Methods: This study was an observational study on natural state. A total of 109 hospitalized patients meeting the diagnostic criteria for depressive episode, single or recurrent depressive disorder in ICD-10, aged ≥60, were enrolled in this study. Cytokines and immune factors were detected and meanwhile HAMD and HAMA were used to estimate depression and anxiety and each sub index of patients. The detection and assessment were completed at baseline and the end of treatment for 4 weeks. Results: The values of IL-1β, IL-6, IL-10 and TNF before treatment and IL-1β after treatment of the patient group were higher than those of the control group. In the patient group, the values of IL-6 , IL-10 and TNF after treatment were lower than those before treatment, and all the differences were statistically significant (P＜0.001 and P＜0.05).The values of CRF and C3 of the patient group after treatment were significantly lower than those before treatment, and the values of IgG and IgM were significantly higher than those before treatment, and all the differences were statistically significant(P＜0.001 and P＜0.05).At baseline of the patient group, positive correlations were found between IL-6 and sleep disorder, IL-8, IL-10 and diurnal variation, TNF and anxiety/somatic, HAMA total score, mental anxiety (P＜0.05 and P＜0.001). In the patient group, before and after treatment, the change of IL-8 concentration was negatively related with the reductive scores of retardation factor of HAMD, and the change of TNF concentration was positively related with the reductive scores of mental anxiety factor and somatic anxiety factor of HAMA (P＜0.05). Conclusion: In the elderly patients with depression, there is a cytokine mediated immune dysfunction, which may be mediated by humoral immune response. The change of TNF concentration may be an indicator of the efficacy of depression in elderly patients.

Abstract:Objective: To investigate the effect of preoperative oral nutrition supplement (ONS) on the complication and perioperative nutritional status of the patients with esophageal cancer who had nutritional risk. Methods: A total of 60 patients with esophageal cancer with nutritional risk screening (NRS)2002≥3 who underwent surgery were randomly divided into the study and control groups. The study group received one week ONS (900 kcal/day of calories and 32 g/day of protein) before surgery and the control group received common semifluid diet only. The preoperative baseline data and postoperative indicators including postoperative complications, volume of drainage, time to exsufflation, albumin and blood transfusion, hospitalization duration and expense and the perioperative serum nutritional indicators (including levels of serum albumin, prealbumin, transferrin and retinol-binding protein before and 1, 3, 5, 7 d after surgery) were recorded. Results: Compared with the control group, the total complication rate (16.7% vs. 40.0%, P＜0.05) and the pulmonary complication rate (6.7% vs. 30.0%, P＜0.05) were significantly reduced in the study group. The perioperative albumin infusion ［(58.0±14.9) mL vs. (76.5±37.9) mL,P＜0.05］, postoperative hospitalization durations ［(15.7±4.3) d vs. (18.8±5.1) d, P＜0.05］ and total hospitalization time ［(22.7±5.7) d vs. (25.8±5.1) d, P＜0.05］ and expense ［￥(66 290.8±9 922.3) vs. ￥(72 491.8±12 535.1), P＜0.05］ were all significantly decreased in the study groups. The level of serum nutritional indicators including albumin, prealbumin, transferrin, retinol-binding protein fell significantly after operation, came to the deepest point on the 3th day after operation and gradually rose after that. However, they were still significantly lower than basal level on the 7th day. The prealbumin level of the study group on the 1th, 3th, 5th, and 7th day after operation were all significantly lower than those of the control group (P＜0.05), which was the same as to the retinol-binding protein except on the 5th day (P＜0.05). Conclusion: One week ONS before operation can effectively promote the nutritional status and postoperative rehabilitation of patients, reduce the incidence of postoperative complications, shorten hospital stay and decrease the expense.

Abstract:Objective: To investigate the value of monitoring the pulse oxygen saturation(SpO2) during walking in the stable chronic obstructive pulmonary disease(COPD) patients. Methods: Twenty-three patients with stable COPD were selected, 20 healthy adults without previous respiratory disease were selected as the control group. SpO2 were monitored during treadmill exercise. The differences between the two groups were compared. The correlation between SpO2 and pulmonary function, COPD assessment test (CAT) score, modified medical research council dyspnea scores(mMRC) and the number of acute execration in the past year were analyzed. Results: ①The nadir SpO2 of the COPD group during the exercise(SpO2ML) was lower than the control group during the exercise with highly significant differences (P<0.05). ② SpO2ML was associated with forced expiratory volume in the first second (FEV1), forced vital capacity (FVC), FEV1%pred and was no significant correlation between FEV1/FVC, maximum ventilatory volume per minute(MVV) and diffusion capacity for carbon monoxide per liter of alveolar volume (DLCO/VA). The acute execration during the past 1 year was not associated with SpO2ML. ③ The CAT score had no significant correlation between the rest SpO2, SpO2ML and ΔSpO2. The mMRC score was associated with SpO2ML, ΔSpO2 and had no significant correlation with the rest SpO2. Conclusion: ①COPD patients are more likely to be exercise-induced hypoxemia. The worse the lung function, the more significant decline in SpO2. ②SpO2ML could be a predictive indicator of the COPD management when the cardiovascular disease was ruled out, and will be a clinical indicator for COPD screening.

Abstract:Objective：To explore the diagnostic value of zinc transporter 8 autoantibody (ZnT8A) and the relationship between ZnT8A and human leukocyte antigen (HLA)-A genotype in the patients with type l diabetes mellitus (T1DM) in Jiangsu Province. Methods：A cross-sectional and case-control study was carried out. The glutamic acid decarboxylase antibody (GADA)，protein tyrosine phosphatase-2 antibody (IA-2A)，insulin antibody (IAA)，and ZnT8A were detected by radioimmunity ligand assay and the HLA gene polymorphism of HLA-A was examined with PCR-SSO method in T1DM and healthy controls (HC). Results：①Among 140 T1DM cases，46.4% (65/140) had positive ZnT8A，while the positive rate in 102 healthy controls was 1.0% (1/102). The positive rate of ZnT8A was significantly higher in T1DM than that in HC (P＜0.05). ②On the base of testing GADA，IAA and IA-2A，the positive rate of at least one antibody was 70.7%(99/140) and it increasd to 86.4% (121/140) when ZnT8A was further tested (P＜0.05). ZnT8A in the other three antibody-negative patients still had a positive rate of 15.7%. ③HLA-A*24 in the T1DM group was significantly higher than that in the healthy control group (P＜0.05，OR=1.654)，while A*33 decreased significantly (P＜0.05，OR=0.161) compared with the healthy control group. ④HLA-A*02 was significantly higher in the ZnT8A-positive group than in the ZnT8A-negative group (P＜0.05，OR=0.318)，but no significant difference was found after adjusting for the effect of course of disease，age of onset，and BMI. Conclusion：The positive rate of antibody increase by 15.7% in T1DM patients when ZnT8A is further tested. ZnT8A can improve the diagnostic sensitivity. No relationship between HLA-A and ZnT8A was found.

Abstract:Objective：To assess the association between -1562C/T polymorphism of the matrix metalloproteinase-9 (MMP-9) promoter and chronic obstructive pulmonary disease (COPD) susceptibility using a comprehensive meta-analysis.Methods：We searched for all case-control studies on the association between MMP-9-1562C/T and COPD susceptibility using the PubMed，EMBASE，FMJS，and CNKI.Additional studies were identified by a hand search of references of original studies or review articles on this topic.Odds ratios and 95% confidence intervals were calculated by using the random-effects model and fixed-effects model.Publication bias test，subgroup analysis，and sensitivity analysis were also performed.Results：A comprehensive meta-analysis was conducted on 13 case-control studies comprising 2 259 cases and 2 539 controls.Results showed that -1562C/T polymorphism of the MMP-9 promoter was associated with the risk for COPD (dominant model：OR=1.40［1.03，1.91］，P=0.03).In addition，MMP-9-1562C/T polymorphism increased risk for COPD among Asians (dominant model：OR=1.80［1.04，3.11］，P=0.04).All the studies included in this meta-analysis were reliable and without publication bias.Conclusion：These findings indicate that -1562C/T of the MMP-9 gene had an association with COPD susceptibility，and playing a role in dominant gene model.

Abstract:Objective：To explore the association between DENND1A polymorphisms and polycystic ovary syndrome (PCOS).Methods：The relevant studies were identified through a search of PubMed，Excerpta Medica Database (Embase)，etc.until Jan 2016.The association between DENND1A polymorphisms and polycystic ovary syndrome risk was assessed by odds ratios(ORs) together with their 95% confidence intervals(CIs).Results：A total of eight studies were eventually identified for DENND1A. We found a significant association between DENND1A rs10818854，rs2479106 and rs10986105 polymorphisms and PCOS susceptibility in overall population.In subgroup analysis，similar results were found in Asian［OR(95%CI)＞1.00，P＜0.05］，while in Caucasian，no significant association was detected between rs2479106 and PCOS(P＞0.05).Conclusion：The present meta-analysis suggests that DENND1A rs10818854，rs2479106，rs10986105 polymorphisms may be associated with polycystic ovary syndrome susceptibility，but there are ethnic differences.All these three polymorphisms are risk factors for Asians：while in Caucasian，rs2479106 is not related to polycystic ovary syndrome susceptibility.Further large and well-designed studies are needed to confirm this association.