ObjectiveTo evaluate the influence of chronic obstructive pulmonary disease on the risk of intrathoracic operation.Methods35 patients associated with chronic obstructive pulmonary disease undergoing intrathoracic operation were analyzed retrospectively.ResultsPostoperative acute and chronic respiratory failure occurred in 4 patients respectively, resulting in 6 deaths.ConclusionThe indication of patients associated with chronic obstructive pulmonary disease for intrathoracic operation should be critically scrutinized, which may include FVC>60% and MVV>40% or FEV1>40%.

A method was developed for determining residual narasin in chicken tissues by HPLC with post-column derivatization. The samples were extracted with iso-octane. Further cleanup was performed on LC-si cartridge after centrifugation. Then the eluent was dried by nitrogen and residues were dissolved in methanol, water mixture (90∶ 10 v/v). The samples were analyzed on an Inertsil ODS-3 C18 column with a mixture of methanol-acetic acid-water as the mobile phase and vanillin as the derivatization reagent. The detection wavelength was 520 nm. The samples were quantified with the external standard calibration curve method. The limit of detection and the limit of quantification for narasin in chicken tissues were 6.0 μg/kg and 20 μg/kg, respectively. The average recoveries of narasin in chicken tissues were 76.4 %-93.1 %, the intra-assay relative standard deviations were 2.6 %-8.9% and the inter-assay relative standard deviations were 4.7%-9.7% at spiked levels of 20-1800 μg/kg. There was a good linear correlation (the calibration coefficient is above 0.9993) between the peak areas and concentration of narasin in the range of 70-10000 μg/L.

For the purpose of resolving such current problems as low inclusion ratio and high exit ratio in the single-disease clinical pathway management,the paper proposed an idea of clinical pathways grouping management,in line with DRG management and based on traditional clinical pathway management practice and China' s conditions.Proposed in the paper are such measures for government service procurement,as quality control mechanism,price negotiation mechanism,performance-based payment mechanism,and supervision mechanism.These measures are designed to provide incentives for medical institutions and medical workers,control irrational rise of medical expense,optimize expense makeup,and improve both quality of care and patient satisfaction.Thanks to the China Rural Health Development Project in place in Henan province where DRG-based clinical pathway grouping management and government service procurement are in trials,the following outcomes are achieved.Namely,significant expansion of diseases coverage and population coverage,efficient control of per-patient/per-visit expenses,and a steady rise of surgery and treatment expenses which embody value of service of medical workers.Other benefits include 50％ to less than 30％drop of drugs proportion,gradual optimization of medical expenses makeup,obvious improvement of quality of care.These measures have made hospitals,insurance providers and patients satisfactory.

Objective To investigate the expression of SOX2 in gastric carcinoma and to analyze its relationship with clinicopathological features.Methods Immunohistochemistry method was used to detect the level of SOX2 in 67 cases of gastric carcinoma group and 30 cases of normal gastric mucosa group.Results The positive expression rate of SOX2 in gastric carcinoma group (52.24 ％) was obviously lower than that in normal gastric mucosa group (93.33 ％) (x2 =16.326,P ＜ 0.01).The SOX2 expression was significantly correlated with differentiation,the depth of invasion,lymph node metastasis and TNM stage of the tumor (all P ＜ 0.05).Conclusions The low expression of SOX2 may contribute to the early carcinogenesis of gastric carcinoma,and the expression level of SOX2 is closely related with lymph node metastasis and TNM stage.The expression level of SOX2 is a useful marker for predicting the prognosis of gastric carcinoma.

Aim To investigate the effect of hydrogen sulfide on hepatic lipid accumulation in obese mice. Methods C57 BL/6 J mice were randomly divided into control group, model group, and NaHS group. The mice of the control group were fed with normal diet. The mice of the model group and the NaHS group were fed with high-fat diet. From the thirteenth week, the mice of NaHS group were injected intraperitoneally with NaHS (H2S donor) in a dose of 50 μmol·kg-1 per day for 4 weeks and the mice of the model group were injected with the same volume of saline. All mice were sacrificed at the end of the 16th week. The tis-sues of liver were homogenized and centrifugated. The supernatants were used for the determination of triglyc-eride and cholesterol in liver. The morphology of liver was tested by H&E staining. Liver lipid accumulation was determined by oil red staining. Total RNA was ex-tracted from frozen tissue of liver. PCR was used to de-tect CPT-1 , FAS gene expression and ELISA method was used to detect CPT-1,FAS activity in mice liver. Results The body weight of the mice from NaHS group and model group was bigger than that of the mice from control group. Compared with the model group, the body weight of the mice from NaHS group was less;the content of triglyceride and cholesterol in liver was lower; the degree of liver tissue pathological changes and lipid accumulation were alleviated; CPT-1 expres-sion and activity were increased; FAS expression and activity were decreased. Conclusions These data in-dicate that hydrogen sulfide can reduce the lipid con-tent of liver tissue in obese mice and alleviate fatty liv-er. The mechanism may be associated with the in-creased expression of CPT-1 and the decreased expres-sion of FAS in liver.

AIM: To evaluate the effect of GYY4137, a novel hydrogen sulfide (H2S) donor, on cytosolic lipid decomposition in mouse primary steatosis hepatocytes.METHODS: Oleic acid (OA) was used to induce hepatic steatosis model in vitro.The C57BL/6 mouse primary hepatocytes isolated and cultured by 2-step in situ perfusion were divided into 4 groups: the cells in control group were incubated with normal medium for 54 h;the cells in model group were incubated with OA at 1.2 mmol/L for 48 h followed by serum-free phenol red-free RPMI-1640 for 6 h;the cells in H2S group or DL-propar-gylglycine (PAG;an inhibitor of cystathione γ-lysase, inhibiting H2S synthesis) group were incubated with OA at 1.2 mmol/L for 48 h followed by serum-free phenol red-free RPMI-1640 which contained 1 mmol/L GYY4137 or 200 μmol/L PAG for 6 h.The glycerin release and the protein expression of hormone-sensitive lipase (HSL) in the cells were mea-sured.RESULTS: Compared with model group, the glycerin release and the protein expression of phosphorylated HSL (p-HSL) in H2S group decreased significantly, while those increased significantly in PAG group.CONCLUSION: In steatosis hepatocytes, exogenous H2S possibly decreases cytosolic lipid decomposition by decreasing the protein level of p-HSL.

[Abstract] Objective： ： To explore miR-103a-3p expression in the tumor tissues and the serum of breast cancer patients, and its role and mechanism in breast cancer development. Methods: Pathologically confirmed 31 cases of tumor tissues and 21 cases of para-cancerous tissues resected at Department of Oncological Surgery of the Second Affiliated Hospital of Hainan Medical University (Haikou, China) from March 1, 2017 to August 31，2017 were collected for this study; in addition, serum samples from 38 breast cancer patients and 22 healthy subjects as well as the breast cancer cell lines MCF-7 and MDA-MB-231 were used in this study. pHBLV-U6-Luc-T2A-Puro and PLL3.7 lentivirus were applied to knock down miR-103a-3p and PDK4 in MCF-7 and MDA-MB-231 cells, respectively. qPCR and Western blotting were performed to examine the mRNA and protein expressions of miR-103a-3p and PDK4 in tissues and serums of breast cancer patients as well as the in cell lines, respectively; CCK-8 assay was applied to detect the proliferation of MCF-7 and MDAMB-231 cells; Olympus AU5400 was applied to detect the glucose consumption and lactate production in indicated cell line. Results： ： miR-103a-3p was significantly decreased in tumor tissues compared with the paracancerous tissues (P<0.01). miR-103a-3p knockdown activated the glucos consumption and lactate production (all P<0.01), increased the PKD4 expression (P<0.01) in MCF-7 and MDAMD-231 cells, and promoted the proliferation of MCF-7 and MDA-MB-231 cells (P<0.01). Furthermore, knockdown of PDK4 suppressed the glucose consumption, lactate production and proliferation in MCF-7 and MDA-MB-231 cells with miR-103a-3p silencing (all P<0.01). Conclusion： ：In the breast cancer, miR-103a-3p inhibited the proliferation of breast cancer cells through down-regulation of PDK4 and PDK4-mediated aerobic glycolysis.

To construct the secretive prokaryotic shuttle expression plasmid pBCG-SP-HSP65, the signal peptide sequence of antigen 85B amplified from Bacillus Calmette-guérin (BCG) genome by PCR and the whole HSP65 DNA sequence of human M. tuberculosis obtained from the plasmid pCMV-MTHSP65 by PCR were cloned into the plasmid pBCG-2100 under the control of the promoter of Heat Shock Protein 70 (HSP70) from human M. tuberculosis. Recombinants were electroporated into Mycobacterial smegmatis and induced by heating. Results of the induced expression were detected by SDS-PAGE and the biological activity of the expressed protein was tested by Western-blot analysis. Results showed pBCG-SP-MTHSP65 was constructed successfully and confirmed by restriction endonuclease analysis, PCR detection and DNA sequencing analysis. After it was electroporated into Mycobacterial smegmatis and induced by heating, the percentage of expressed 65kD protein in Mycobacterial smegmatis detected by SDS-PAGE was 20% in total bacterial protein. But the percentage of expressed 65kD protein in recombibinant Mycobacterial smegmatis was up to 34.46% in total bacterial protein and 68.56% in the total protein of cell lysate supernants, Which demonstrated the recombinant HSP65 gene could express in recombinant with high efficiency and the expressed proteins were mainly soluble. Western-blot showed that the secretive proteins could specially combine with antibody against human M. tuberculosis HSP65. Orally, pBCG-SP-HSP65 was successfully constructed; HSP65 gene could express in Mycobacterial smegmatis with high efficiency via it. And the expressed proteins possess the biological activity. So it provids experimental evidence for the application of the recombinant Mycobacterial smegmatis and the development of the vaccine against tuberculosis.
Bacterial Proteins ; biosynthesis ; genetics ; Chaperonin 60 ; Chaperonins ; biosynthesis ; genetics ; Escherichia coli ; genetics ; metabolism ; Humans ; Mycobacterium smegmatis ; genetics ; metabolism ; Plasmids ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics

AIM:To evaluate the effect of exogenous hydrogen sulfide ( H2 S) from GYY4137 on lipophagy in mouse primary hepatocytes .METHODS:The C57BL/6 mouse primary hepatocytes isolated and cultured by 2-step in situ perfusion method were divided into 4 groups:the cells in control group were incubated with normal medium; the cells in model group were incubated with 1.2 mmol/L oleic acid (OA) for 48 h;the cells in H2S group or propargylglycine (PAG) group were incubated with 1.2 mmol/L OA for 48 h followed by serum-free phenol red-free RPMI-1640 medium which con-tained 1 mmol/L GYY4137 or 200 μmol/L PAG for 6 h.The cells were collected to conduct immunofluorescence staining of LC3 and photography under fluorescence microscope , phase-contrast microscope or transmission electron microscope . The protein expression of LC 3-Ⅰ/Ⅱin the hepatocytes was determined by Western blot .RESULTS:In contrast with the model group, the fluorescent particles of LC3, the protein expression of LC3, the number of autophagic lysosome and vacuoles in hepatocytes in H 2 S group increased .CONCLUSION: In steatosis hepatocytes , exogenous H 2 S promotes the lipophagy .

Objective To investigate the relationship between the initial change of thyroglobulin (Tg) and clinical outcome in differentiated thyroid carcinoma (DTC) patients with pulmonary metastases after 131I treatment. Methods A total of 69 DTC patients with pulmonary metastases from January 2010 to December 2015 were retrospectively analyzed. Patients were divided into 3 groups according to the variation of Tg: groupⅠ(Tg declined≥ 50%), groupⅡ(Tg declined＜50% or Tg increased＜10%), and group Ⅲ (Tg increased ≥10% ). The follow-up time was (49.2 ± 9.3) months. Clinical outcomes were divided into remission, stable disease and progressive disease according to the serum test and imaging results. Results The percentage of group Ⅰ, Ⅱ, Ⅲ patients was 44.9% (31/69), 40.6% (28/69), and 14.5% (10/69) respectively. Results of follow-up showed 19.4% (6/31) patients achieved remission and 80.6% (25/31) had stable disease in groupⅠ. There were 10.7% (3/28) patients with remission, 60.7% (17/28) with stable disease and 28.6% (8/28) with progression disease in groupⅡ. All patients showed progressive disease in groupⅢ. The clinical outcome was related to the variation of Tg after 131I treatment (P＜0.01). Conclusions Initial Tg after 131I treatment could be a predictor to the outcome of patients. The increased Tg level indicates a high possibility of 131I refractory disease.