Objective To analyze the results of sputum culture from smear-positive pulmonary tuberculosis cases after treatment for 2 months.Methods Sputum specimens from newly diagnosed 147 smear-positive pulmonary tuberculosis cases after treatment for 2 months were fast cultured by BacT/ALERT 3D.Poeitive cultures were further detected by HAIN test in Zhejiang Provincial Center for Disease Control and prevention and compared with L-J meditun.Results Mycobacteria were found in 45 specimens by Bact/ALERT 3D in contrast to 22 by Lonswtein-Jenson method.The median time was 19.3 days for the whole Bact/ALERT 3D test while the whole procedure would cost 3 months for traditional L-J test.Conclusion Results from sputum smear could not reflect tuberculosis progress properly while BacT/ALERT 3D fast culture would help to find drug resistant patients promptly and accurately.

Objective To research the effect of alimentary tract reconstruction after gastrectomy on the treatment of type 2 diabetes mellitus(non-insulin-dependent diabetes mellitus, NIDDM). Methods From January 2005 to January 2008, perioperative blood glucose level and insulin therapy in 24 gastric cancer or periampullary carcinoma patients with non-obesity type 2 diabetes were retrospectively analyzed. These patients underwent different alimentary tract reconstruction, including 8 patients for Billroth I, 10 for standard Whipples operation, 6 for esophageal Roux-en-Y jejunostomy after total gastrectomy. Glucose level and insulin dosage of thease patients were compared. Results In the patients underwent Billroth I operation, change of blood glucose level before and after operation was not significant(P＞0.05). The level of blood glucose in patients underwent Whipples operation and total gastrectomy were significant changed 1 and 2 months after the operation (P<0.001). In Billroth I group, 5 patients used insulin to control hyperglycemia preoperatively. After the operation, 2 patents maintained preoperative insulin dosage, 2 had to increase the use of insulin and 1 decreased the insulin dosage. In Whipples group, 6 patients used insulin preoperatively. Interestingly, 4 patents needed less insulin to control hyperglycemia and 2 were free of insulin dependance. In total gastrectomy group, there were 4 patients using insulin to control hyperglycemia. After the operation, 2 patents needed less insulin dosage and 2 stopped using insulin. Conclusions Both Whipples operation and total gastrectomy lead to decreased blood glucose level in NIDDM patients and less need of insulin.The effect of some types of alimentary tract reconstruction after gastrectomy on treatment of type 2 diabetes mellitus is assertive.

Objective To investigate the therapeutic effect and possible mechanism of adenosine A2A receptor agonist (CGS21680) combined with bone marrow mesenchymal stem cells (BMMSC) transplantation in acute liver failure (ALF).Methods Fifty male C57BL/6 mice, 6-8 weeks old, were fed with standard diet for 1 week and randomly divided into 5 groups according to random number table: healthy control group (n=6), model group (n=11), BMMSC group (n=11), CGS21680/BMMSC group (n=11) and CGS21680 group (n=11).Except healthy control group, the other mice were injected with D-GalN and lipopolysaccharide (LPS) to establish ALF model.Ten hours later, CGS21680/BMMSC group and CGS21680 group were injected intraperitoneally with adenosine A2A receptor agonist CGS21680 (2.1 mg/kg).In addition, the BMMSC group and CGS21680/BMMSC group were injected BMMSC (1×10.6) through tail vein.After 24 hours, pathological changes of liver tissue was observed by hematoxylin and eosin staining.The change of proportion of mouse splenic Treg among CD4+T lymphocytes was detected by flow cytometry.Toll-like receptor (TLR)4 and nuclear factor (NF)-κB expression levels in liver tissue were detected by real-time fluorescence quantitative polymerase chain reaction (PCR) and Western blot.One-way analysis of variance (one-way ANOVA) and SNK-q test was conducted for data analysis.Results Serum IL-6 levels were (23.67±2.97) pg/mL in healthy control group, (151.47±6.03) pg/mL in model control group, (72.10±3.74) pg/mL in BMMSC group, (53.35±2.50) pg/mL in CGS21680/BMMSC group and (84.85±3.25) pg/mL in CGS21680 group.The differences between healthy control group and the other 4 groups were all statistically significant (t=46.02, 25.51, 19.58 and 34.03, respectively, all P<0.01).Serum TNF-ɑ levels were (24.62±3.19) pg/mL in healthy control group, (102.25±2.10) pg/mL in model control group, (54.71±2.23) pg/mL in BMMSC group, (42.20±4.72) pg/mL in CGS21680/BMMSC group and (81.76±3.50) pg/mL in CGS21680 group.The differences between healthy control group and the other 4 groups were all statistically significant (t=46.49, 19.97, 7.72 and 29.57, respectively, all P<0.01).The differences of spleen Treg proportion in healthy control group were statistically significant compared with model control group, BMMSC group, CGS21680/BMMSC group and CGS21680 group (t=51.67, 12.22, 5.91 and 18.21, respectively, all P<0.01).The differences of TLR4 mRNA levels of liver tissue in healthy control group were statistically significant compared with model control group, BMMSC group, CGS21680/BMMSC group and CGS21680 group (t=26.31, 21.33, 13.24 and 27.14, respectively, all P<0.05).The differences of NF-κB mRNA level of liver tissue in healthy control group were statistically significant compared with model control group, BMMSC group, CGS21680/BMMSC group and CGS21680 group (t=16.56, 16.34, 7.83 and 13.11, respectively, all P<0.05).The differences of TLR4 protein level in liver tissue of healthy control group were statistically significant compared with model control group, BMMSC group, CGS21680/BMMSC group and CGS21680 group (t=35.60, 10.38, 6.05 and 18.02, respectively, all P<0.05).The differences of liver NF-κB protein level in the healthy control group were statistically significant compared with model control group, BMMSC group, CGS21680/BMMSC group and CGS21680 group (t=10.80, 7.30, 4.61 and 13.24, respectively, all P<0.05).Conclusions Adenosine A2A receptor agonist combined with BMMSC can significantly up-regulate the proportion of Treg cells in acute liver failure mice and inhibit the TLR4/NF-κB pathway activation, with both coordinated regulation, and further inhibit the liver inflammation.

Objective To investigate whether the effect of transient high glucose on inflammatory factors expression could be continuous in rat glomerular mesangial cell,and its relation with histone methylation modification.Methods Rat glomerular mesangial cells (HBZY-l) were divided into three groups:the high glucose group (25.0 mmol/L glucose),the hypertonic group (MA,5.5 mmol/L glucose+ 19.5 mmol/L mannitol) and the normal-glucose control group (5.5 mmol/L glucose),which were cultured for 24 h respectively.All 3 groups were then changed with normal-glucose medium to culture for 24 h,48 h and 72 h.Their protein,mRNA and supernatant were harvested.The protein expressions of mono-methylation of H3 lysine 4 (H3K4mel) was measured by Western blotting,and the mRNA expressions of NF-κB subunit p65 and set7/9 were determined by real timequantitative PCR.The expression of monocyte chemoattractant protein 1 (MCP-1) and vascular cell adhesion molecule 1 (VCAM-1) were detected by enzyme-linked immunosorbent assay.Results (1)Compared with those in normal control group,the expressions of H3K4mel protein and set7/9 mRNA were first up-regulated in high glucose group,then gradually down-regulated in the following 48 h normal-glucose medium (as compared with those at 0 h,all P ＜ 0.05).At 72 h there was no statistic difference between high glucose group and normal control group (all P ＞ 0.05).(2) Compared with those in normal control group,the up-regulated p65 mRNA,VCAM-1 and MCP-1 sustained at least for 72 h in high glucose group.Conclusions Transient high glucose can induce persistent inflammatory factors expression in rat glomerular mesangial cells,which may via histone modification.

Objective To evaluate the effect of recombinant human tumor necrosis factor receptor type Ⅱ:IgG Fc fusion protein (rhTNFR:Fc,trade name Etanercept) combined with methotrexate on levels of interleukin-17A (IL-17A) and tumor necrosis factor-α (TNF-α) in the serum and mononuclear cells of patients with moderate to severe plaque psoriasis.Methods A total of 30 patients with moderate to severe plaque psoriasis were enrolled from Department of Dermatology of Tenth People's Hospital of Tongji University between August 2014 and February 2016,and then were randomly and equally divided into Etanercept group and Etanercept + methotrexate group.The treatment lasted 24 weeks.Fifteen healthy blood donors served as healthy control group.Enzyme-linked immunosorbent assay (ELISA) and real-time quantitative PCR were performed to measure the serum levels and mRNA expression of IL-17A and TNF-α,respectively,in the patients of the above two groups before and after the treatment.Results Before the treatment,the serum levels of IL-17A and TNF-ct,as well as the mRNA expression of IL-17A and TNF-α in peripheral blood mononuclear cells (PBMCs),were all significantly higher in all the patients than in the healthy controls (all P ＜ 0.05).After the treatment,compared with the Etanercept group,the Etanercept + methotrexate group showed significantly lower serum levels of IL-17A (142.67 ± 14.82 vs.163.54 ± 23.18,P ＜ 0.05) and TNF-α (70.07 ± 25.02 vs.91.98 ± 14.62,P ＜ 0.05),as well as lower mRNA expression of IL-17A (1.12 ± 0.33 vs.1.56 ± 0.77,P ＜ 0.05) and TNF-α in PBMCs (2.50 ± 1.04 vs.3.61 ± 2.14,P ＜ 0.05).Conclusion Etanercept combined with methotrexate is superior to Etanercept alone in the treatment of psoriasis,and can reduce treatment duration and improve therapeutic effect,likely by down-regulating the expression of IL-17A and TNF-α.

Objective To investigate the effects of bone marrow-derived stem cells (BMMSC)on the expressions of inflammatory cytokines and Toll-like receptor (TLR)4 pathway in primary hepatic stellate cells (HSC)with direct and indirect contact coculture system.Methods Purified HSC were separately treated with LPS in the concentrations of 0,50,100 and 150 g/L for 48 h.Proliferation ratio was tested with cell counting kit-8 to determine the optimal concentration.HSC in LPS were divided into three groups,including HSC alone group,cocultured with BMMSC at 1∶1 group and cocultured with transwell group at the optimal concentration.The supernatants were collected to detect the concentrations of interleukin-6 (IL-6)and tumor necrosis factor (TNF)-α.Cells were further divided into seven groups, including BMMSC without LPS group,HSC without LPS group,BMMSC with LPS group,HSC with LPS group,BMMSC in transwell system group,HSC in transwell system group,all cells in transwell system group and direct contact system group.The mRNA expressions ofα-smooth muscle actin (α-SMA) and collagen I were detected by quantitative real-time PCR,and protein expressions of TLR4,myeloid differentiation factor 88 (MyD88)and nuclear factor-κB (NF-κB)were analyzed by Western blot.Data were analyzed with one-way ANOVA analysis.Results The proliferation rate of HSC in 50,100 and 150μg/L LPS were (129.77±11 .26)%,(162.90±13.15 )% and (55 .12 ±11 .6)%,respectively compared with HSC without LPS group.The differences were statistically significant (t =5 .91 ,10.70 and 8.65 , respectively,all P <0.05).The concentrations of IL-6 and TNF-αin HSC alone group,directly/indirectly cocultured group were (252.02 ±30.94)ng/L and (148.00 ± 10.27 )ng/L,(88.52 ±6.61 )g/L and (72.63±5 .54)ng/L,(103.74±7.14)ng/L and (81 .79 ±6.92 )ng/L,respectively.The differences between HSC alone group and directly/indirectly cocultured group were significant (t=12.66 and 15 .82, 11 .81 and 12.34,respectively,all P < 0.05 ).The directly and indirectly cocultured groups were significantly different (t=3.83 and 2.53,respectively,both P <0.05 ).The mRNA expressions of α-SMA and collagen I in HSC with LPS were remarkably increased compared with HSC without LPS (t =14.16 and 11 .84,respectively,both P <0.05 )and reversed by cocultured with BMMSC (t =11 .98 and 4.47,respectively,P <0.05).All cells in transwell group expressed moreα-SMA and collagen I than the direct contact group (t=3.70 and 3.19,respectively,P <0.05 ).The TLR pathway associated protein expressions,TLR4,MyD88,and NF-κB in HSC in transwell group were significantly down-regulated compared with HSC with LPS group.And all cells in transwell system had higher level of protein expressions compared with direct contact system (P < 0.05 ).Conclusions BMMSC are effective in inhibiting HSC activation and inflammatory cytokines excretion,which may be modulated through TLR4 pathway and cell to cell contact.

Biochemical analysis assays based on colorimetric methods using gold nanoparticles have many advantages including high sensitivity, good selectivity, naked-eyes readout and complex instruments free. These methods have good prospects in applications. The biomolecule assay is highly relative with human health. This review mainly focuses on colorimetric assays applying gold nanoparticles for biomolecules detection.

Objective To analyze the effects of valproic acid(VPA),a histone deacetylase(HDAC)inhibitor,on macrophage polarization in?duced by paraquat(PQ)or lipopolysaccharide(LPS). Methods Mouse RAW264.7 cells were cultured at 37℃with 5%CO2,passaged,and then given one of the following treatments:(1)PQ;(2)PQ+VPA(classⅠandⅡa HDAC inhibitor);(3)PQ+apicidin(classⅠHDAC inhibitor);(4)PQ+MC1568(classⅡa HDAC inhibitor);(5)LPS;(6)LPS+VPA;(7)LPS+apicidin;(8)LPS+MC1568. The cells and culture supernatants were harvested after 8 h of treatment. RT?PCR,ELISA,and flow cytometry were conducted to assess the expression levels of macrophage phenotyp?ic markers. Results Both PQ and LPS skewed the macrophage functional polarity toward proinflammatory phenotype. VPA,apicidin,and MC1568 all inhibited PQ?and LPS?induced macrophages polarizing toward pro?inflammatory phenotype ,but the inhibitory effects were different in some ways. Conclusion VPA inhibits the proinflammatory function of macrophages induced by PQ and LPS ,but the effect of VPA on PQ?and LPS?induced macrophages has its own characteristics.

Objective: To explore the effectiveness of KeyPort access in transanal endoscopic mircrosurgery (TEM). Methods: A descriptive case series study was performed. Clinicopathological data of 20 patients undergoing KeyPort access TEM in Beijing Chaoyang Hospital of Capital Medical University from December 2016 to April 2018 were collected and analyzed retrospectively. Procedure of KeyPort access TEM: general anesthesia or combined spinal epidural anesthesia (CSEA); lithotomy or prone jack-knife position; anal dilation; placement of the KeyPortaccess; connection of TEM pneumoperitoneum device, light source and imaging equipment; placement of 5 mm dedicated endoscope; insufflation of CO2 with pressure of 1.6-2.0 kPa (12-15 mmHg); after rinsing the intestinal lumen, circular resection marginlabeled by the needle-shaped electrocautery;electric coagulation or ultrasonic knife used to perform a full-thickness resection with a 0.5 cm-1 cm margin along the marking line. Indications of KeyPort access TEM: (1) benign large sessile polyps which were difficult to resect under colonoscopy; (2) submucosal lesions with diameter <2 cm; (3) Tis and T1 stage rectal carcinoma without lymph node metastasis; (4) palliative resection of T2 stage rectal carcinoma without lymph node metastasis. Contraindications: (1) accompanying serious diseases without the tolerance of anesthesia and operation; (2) distance from lesion to anal verge >20 cm. Results: There were 10 males and 10 females with age of (63±15) years old and BMI of (24.5±3.3) kg/m². The diameter of the lesions was (2.0±1.3) cm, and the distance from lesion to anal verge was (6.2±2.2) cm. One patient had 3 lesions at different positions in rectum with diameters of 0.5 cm, 0.5 cm, and 1 cm, respectively. All operations were accomplished through the KeyPort access TEM and no case was transferred to other methods. The duration of surgery was 75 (30-220) minutes; intraoperative blood loss was 10 (0-30) ml. Two patients with rectal anterior wall lesions underwent full-thickness resection of the intestine wall reaching the peritoneal reflex with penetration into the peritonealcavity, and received suture closure immediately. For the patient with 3 rectal lesions, the 1.0 cm lesion received a full-thickness resection and the other 2 lesions received submucosal resection. No postoperative complication occurred. Postoperative pathology showed that there were 1 case of chronic inflammatory lesion, 4 cases of benign tumor, 3 cases of carcinoma in situ, 4 cases of neuroendocrine tumor, 6 cases of pT1 rectal cancer, 2 cases of pT2 rectal cancer (both invading the superficial muscle layer). The median hospital stay was 6 (3-7) days. The postoperative follow-up was (7.2±3.8) months. No postoperative complication or recurrence was observed. Conclusion TEM with KeyPort access is safe, rapid and effective in the treatment of rectal tumors.

Objective The use of the antifungal agent itraconazole and the calcineurin inhibitor tacrolimus alone, together with the above two drugs, was evaluated in vitro for growth of Candida krusei. Whether or not the combina-tion is more effective than the antifungal alone can inhibit the growth of Candida krusei. Methods According to the guidelines of the American Institute of Clinical and Laboratory Standards, 12 Candida krusei strains were divided into blank control group, itraconazole group, tacrolimus group, and combination group by microdilution method, and then 12 the Candida krusei strain was incubated for 24 h and its growth was observed. Results In comparison with the alone use of isotriconazole in Candida krusei, the combined use of itraconazole,its minimal inhibitory con-centration was much less than that of alone, with a maximum multiple of 32 fold, a minimum multiple up to 4 times; the same, In comparison with the alone use of tacrolimus in Candida krusei, the combined use of tacroli-mus, its maximum multiple of 32 times, the lowest multiple of 2 times. Conclusion The combined use of itracon-azole and tacrolimus in 12 strains of Candida krusei displayed a powerful synergistic effect in vitro.