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Based on the rDNA sequence of Pichia pastoris, a multi-copy gene expression vector of transglutaminase (pPICZα-rDNA-mtg) was constructed and transformed to the host strain (pGAP9-pro/GS115) expressing pro peptide, to obtain the co-expression strain pro/rDNA-mtg (GS115). Real-time fluorescence quantitative PCR (qPCR) was used to analyze transglutaminase gene copy number in the 4 positive strains. We further studied the effect of gene copy on the enzyme production of recombinant Pichia pastoris as well as high-density fermentation of higher expression strain in a 3-L fermenter. The mtg copy numbers of the 4 positive strains were 2.21, 3.36, 5.72 and 7.62 (mtg-2c, mtg-3c, mtg-6c and mtg-8c), respectively, and the enzyme production capacity and protein expression level were mtg-3c>mtg-2c>mtg-6c>mtg-8c. Mtg-3c and mtg-6c of high-density fermentation had the highest enzymatic activity and enzymatic activity per unit wet weight in the supernatant of 3.12 U/mL, 52.1 U/g (wet weight) and 2.07 U/mL and 36.5 U/g (wet weight), respectively. In terms of enzyme activity per unit wet weight, mtg-3c is 1.4 times higher than that of mtg-6c. The activity of purified enzyme (mtg-3c) was up to 7.21 U/mL and the protein concentration was 437.2 μg/mL. By analyzing the effect of mtg copy number on the enzyme production of recombinant strains, mtg-3c is suitable for the co-expression of two genes (pro and mtg) in pro/rDNA-mtg, and its enzyme activity is related to higher protein secretion of the strain.
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Objective:To observe the effects of bundled pulmonary rehabilitation on static pulmonary function, cardiopulmonary exercise function and inflammatory factors in patients with moderate or severe stable chronic obstructive pulmonary disease (COPD).Methods:Sixty moderate or severe COPD patients in stable phase from December 2017 to October 2018 in Affiliated Zhongshan Hospital of Dalian University were selected. The patients were divided into control group and rehabilitation group by random digits table with 30 cases each. The patients of control group received routine medicine therapy, and the patients of rehabilitation group received bundled pulmonary rehabilitation besides routine medicine therapy. Pulmonary function, cardiopulmonary exercise function and inflammatory factors in each group were tested before treatment and 3, 6 month after treatment.Results:In control group, there were no statistical differences in the pulmonary function, including the forced expired volume in one second (FEV 1), FEV 1 as percentage of predicted (FEV 1%pred), FEV 1/forced vital capacity (FVC); cardiopulmonary exercise function, including the peak load, peak VO 2, peak VO 2 as percentage of predicted (peak VO 2% pred), peak VO 2/kg, peak VE 3 and 6 months after treatment compared with those before treatment ( P>0.05); in rehabilitation group, there were no statistical differences in the indexes 3 months after treatment compared with those before treatment ( P>0.05), the indexes 6 months after treatment were significantly improved compared with those before treatment and 3 months after treatment ( P < 0.05). In control group, the C-reactive protein (CRP) and interleukin (IL-6) before treatment and 3 months after treatment and tumor necrosis factor-α (TNF-α) before treatment and 3, 6 months after treatment had no significant difference ( P > 0.05); the CRP and IL-6 6 months after treatment were significantly lower than those before treatment and 3 months: (5.48 ± 0.91) mg/L vs. (6.26 ± 0.99) and (6.09 ± 0.97) mg/L, (9.17 ± 1.04) ng/L vs. (10.83 ± 1.79) and (10.29 ± 1.53) ng/L, P < 0.05. In rehabilitation group, the CRP, IL-6 and TNF-α 3 and 6 months after treatment were significantly lower than previous phase: (5.21 ± 0.87) and (5.67 ± 0.91) mg/L vs. (6.15 ± 0.92) mg/L, (8.56 ± 1.17) and (10.03 ± 1.61) ng/L vs. (11.06 ± 2.01) ng/L, (7.16 ± 1.33) and (8.05 ± 1.62) ng/L vs. (8.97 ± 2.05) ng/L, P < 0.05. In rehabilitation group, the CRP, IL-6 and TNF-α before and after treatment were negatively correlated with pulmonary function (FEV 1, FEV 1% pred and FEV 1/FVC) and cardiopulmonary exercise function (peak load, peak VO 2, peak VO 2% pred, peak VO 2/kg and peak VE) ( r=- 0.641 to - 0.884, P < 0.01). Conclusions:Bundled pulmonary rehabilitation in patients with moderate or severe stable COPD can improve pulmonary function and cardiopulmonary exercise function and reduce inflammatory factors.
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Objective:To investigate the molecular mechanism of SUMO specific protease 1 (SENP1) regulating endoplasmic reticulum stress transcription regulator X-box binding protein 1 (XBP1) in the proliferation of liver cancer cells.Methods:The pathological samples of 180 patients with primary liver cancer in the Department of Hepatobiliary Surgery of Inner Mongolia People′s Hospital from January 2012 to January 2020 were collected. The expressions of SENP1 and XBP1 in liver cancer, adjacent tissues and different liver cancer cell lines were detected. The correlation between SENP1 positive expression and clinicopathological features of liver cancer patients was analyzed. Immunofluorescence and flow cytometry were used to detect the effect of SENP1 siRNA on XBP1 and apoptosis. SUMO1 expression on XBP1 surface was detected and the effect of SENP1 siRNA on SUMO formation of XBP1 was detected by immunoprecipitation.Results:The expression levels of SENP1 in liver cancer and adjacent tissues were 16.332±4.371 and 6.840±2.238, with a statistically significant difference ( t=-5.073, P=0.017). The expression levels of XBP1 in liver cancer and adjacent tissues were 6.641±2.482 and 16.051±4.452, with a statistically significant difference ( t=3.592, P=0.032). The expression of SENP1 was correlated with stage ( χ2=6.724, P=0.010) and metastasis ( χ2=6.265, P=0.012). Immunofluorescence staining showed that the expressions of XBP1 in L02 (0.509±0.219), MHCC97-L (0.092±0.022) and HCCLM3 (0.086±0.014) cells were significantly different ( F=6.378, P=0.004), while the expression of XBP1 in MHCC97-L and HCCLM3 cells was significantly lower than that in L02 cells ( P=0.023; P=0.021). The expression levels of SENP1 in L02, MHCC97-L and HCCLM3 cells were 0.109±0.079, 0.802±0.392 and 0.921±0.352, with a statistically significant difference ( F=7.783, P=0.004), while the expression level of SENP1 in MHCC97-L and HCCLM3 cells was significantly higher than that in L02 cells ( P=0.039; P=0.016). After transfection of SENP1 siRNA into MHCC97-L and HCCLM3 cells, the expressions of XBP1 increased (0.462±0.192, t=3.664, P=0.022; 0.524±0.203, t=3.383, P=0.028); the expressions of SENP1 decreased (0.153±0.093, t=2.790, P=0.049; 0.165±0.104, t=3.568, P=0.023). The results of flow cytometry showed that the apoptosis rates of L02, MHCC97-L, HCCLM3, MHCC97-L+ SENP1 siRNA and HCCLM3+ SENP1 siRNA cells were (20.80±3.11)%, (2.02±1.20)%, (0.12±0.01)%, (7.01±1.80)%, (6.20±2.01)%, with a statistically significant difference ( F=1.025, P=0.030). The apoptosis rate of MHCC97-L and HCCLM3 cells was significantly lower than that of L02 cells ( P=0.040; P=0.010), the apoptosis rate of MHCC97-L+ SENP1 siRNA and HCCLM3+ SENP1 siRNA cells was significantly higher than that of MHCC97-L and HCCLM3 cells (both P=0.009). Immunoprecipitation results showed that the expression levels of XBP1 in L02, MHCC97-L, HCCLM3, MHCC97-L+ SENP1 siRNA, HCCLM3+ SENP1 siRNA cells were 11.943±5.043, 7.467±1.903, 2.051±0.913, 9.532±3.012, 8.731±3.102, and SUMO1 expression levels were 10.158±4.005, 5.871±3.075, 1.941± 0.907, 8.658±4.878, 7.169±4.677, and the differences were statistically significant ( F=11.730, P=0.010; F=8.548, P=0.001). The expressions of XBP1 and SUMO1 in MHCC97-L ( P=0.028; P=0.038) and HCCLM3 ( P<0.001; P<0.001) cells were lower than those in L02 cells, XBP1 expression in HCCLM3+ SENP1 siRNA cells was higher than that in HCCLM3 cells ( P=0.001), and SUMO1 expression in MHCC97-L+ SENP1 siRNA cells and HCCLM3+ SENP1 siRNA cells respectively was higher than that in MHCC97-L ( P=0.045) and HCCLM3 ( P=0.039) cells. Conclusion:SENP1 siRNA can promote the apoptosis of liver cancer cells by up regulating SUMO modification of XBP1.
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BACKGROUND@#The purpose of this study is to compare the survival time of non-small cell lung cancer (NSCLC) patients with different organ metastasis. Among all cancers, the morbidity and mortality of lung cancer is the highest worldwide, which may caused by local recurrence and distant metastasis, and the location of metastasis may predict the prognosis of patients.@*METHODS@#A total of 117,542 patients with NSCLC diagnosed between 2010 and 2014 were enrolled from Surveillance, Epidemiology, and End Result (SEER) databases, and the relationship between distant metastasis and survival time was retrospectively analyzed.@*RESULTS@#Of all the 117,542 patients diagnosed with non-small cell lung cancer, 42,071 (35.8%) patients had different degrees of distant metastasis during their medical history, including 26,932 single organ metastases and 15,139 multiple organ metastases, accounting for 64.0% and 36.0% of the metastatic patients respectively. Compared with patients with no metastasis, whose median survival time was 21 months, the median survival time of patients with metastases was 7 months (lung), 6 months (brain), 5 months (bone), 4 months (liver), and 3 months (multiple organ) respectively, and the difference was significant (P<0.001, except liver vs multiple organ P=0.650); Most patients with NSCLC (88.4%) eventually died of lung cancer.@*CONCLUSIONS@#Distant metastasis of NSCLC patients indicates poor prognosis. In NSCLC patients with single organ metastasis, the prognosis of lung metastasis is the best, and liver metastasis is the worst, and multiple organ metastasis is worse than single organ metastasis.
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Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Bone Neoplasms , Mortality , Brain Neoplasms , Mortality , Carcinoma, Non-Small-Cell Lung , Mortality , Pathology , Liver Neoplasms , Mortality , Lung Neoplasms , Mortality , Pathology , Neoplasm Metastasis , Neoplasm Staging , Prognosis , Retrospective StudiesABSTRACT
OBJECTIVE: To analyze carbapenemases genotype of imipenem-resistant Gram-negative bacilli in intensive care unit (ICU) of 3 third grade class A hospitals from Qingdao area, so as to provide reference for drug-resistant bacteria infection prevention and treatment in clinic. METHODS: From Jan. 2013 to Jun. 2016, each 60 strains of imipenem-resistant Klebsiella pneumoniae (IRKP), imipenem-resistant Pseudomonas aeruginosa (IRPA) and imipenem-resistant Acinetobacter baumanii (IRAB) were collected from 3 third grade class A hospitals from Qingdao area. Drug sensitivity test was performed by using Kirby-Bauer method. Phenotypes of carbapenemases were determined by Carba NP trial. PCR was applied to amplify carbapenemase gene; Sanger seqnencing method was adopted for bi-directional sequencing; Blast comparison with GenBank database was conducted. RESULTS: Three kinds of imipenem-resistant Gram-negative bacilli showed high drug resistance to majority commonly used antibiotics as piperacillin, cefazolin, imipenem and cilastatin sodium, gentamicin, etc., but were sensitive to polymyxin B (resistance rate of 0). Among 180 drug-resistant strains, there were 52 strains of class A carbapenems, 13 strains of class B carbapenems and 39 strains of class D carbapenems; the detection rates of them were 28. 89％, 7. 22％ and 21. 67％, respectively. There were 52 strains of KPC-2 gene (IRKP), 4 strains of IMP-1 gene (IRPA), 8 strains of VIM-2 gene (7 strains of IRPA, 1 strain of IRAB), 39 strains of OXA-23 gene (IRAB); the detection rates of them were 28. 89％, 2. 22％, 4. 44％, 21. 67％; all strains were not detected 1MP-2, VIM-1, NDM-1, OXA-24, OXA-58 genes. Results of Blast comparison showed that above detected genes were absolutely homology with the corresponding genes in GenBank database. CONCLUSIONS: Drug resistance of imipenem-resistant Gram-negative bacilli in ICU of 3 third grade class A hospitals is serious in this region, which are nearly no-sensitive to most of commonly used antibiotics in clinic. Main genotypes included KPC-2 (K. pneumoniae), OXA-23 (A. baumanii) and IMP-1 and VIM-2 (P. aeruginosa).
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Objective To investigate antibiotics resistant characteristics and carbapenemases genotype of Acinetobacter baumannii in Intensive Care Unit (ICU),so as to provide theoretical basis for clinical prevention and treatment.Methods Retrospective study was made on 90 non-duplicated clinical isolates of Acinetobacter baumannii,which were collected From January 2013 to January 2014 in three tertiary hospitals of Qingdao.All strains were identified by VITEK2 automated microbiology analyzer;K-B method was used to do drug susceptibility test;polymerase chain reaction (PCR) was used to amplify the OXA-23,OXA-24,OXA-51,OXA-58,KPC-2,VIM,IMP genes,and the positive products of genes were sequenced;the chi-square test was used to compare the difference of the resistance rates.Results The detection rate of multi-drug resistant A.baumannii (MDRAB)and Pan-drug resistant A.baumannii (PDRAB)was 61.11% (55/90) and 17.78% (16/90).In the 32 strains of imipenem-resistant Acinetobacter baumannii,the resistant rates to Cefoperazone/sulbactam,Polymyxin B was lower,while the resistant rates to other drugs tested were more than 85%.The difference of the resistance rates to 9 drugs between imipenem resistant group and Imipenem sensitive group were statistically significant (P≤0.05).PCR result showed: 32 strains detected OXA-51 gene,28 strains detected OXA-23 gene,and 3 strains detected VIM gene,the detection rates of which were 100%,87.50% and 9.38% respectively.All strains were not detected OXA-24,OXA-58,KPC-2 and IMP genes.The sequenced results were absolutely homology with the corresponding genes in genbank.Conclusions The resistance of A.baumannii in ICU is serious in this region,especially imipenem-resistant A.baumannii,which were nearly no-sensitive to most of the drugs commonly used in clinical.The gene existence of carbapenemase and carbapenemase producing is one of the main resistance mechanism of Acinetobacter baumannii to carbapenem antibiotics.OXA-23 was the major genotypes in this region.
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Objective To study the reason of concurrent styloid process syndrome after tonsillectomy and the clinical effect of styloidectomy.Methods Twelve patients with styloid process syndrome after tonsillectomy receiving styloidectomy were retrospectively analyzed.The clinical effect was evaluated.Results Ten patients were operated on unilateral styloid,two patient was operated on bilateral styloid.All patients were followed -up for 6 months to 5 years after operation.All of them were cured.Conclusion The mechanism of styloid process syndrome after tonsillectomy might be related to the stimulation of scar formation in the tonsillar fossa.Styloidectomy ia an effective method of treatment.
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Objective To investigate the prevalence of infertility and related factors in Uygur and Kazak women in Xinjiang Uygur autonomous region (Xinjiang).Methods Questionnaire survey and pelvic examination were conducted among 535 Uygur women and 322 Kazak women at reproductive age who were selected through stratified cluster random sampling in Sansan and Fuhai counties in Xinjiang.The data were analyzed with software SPSS 17.0.Results The prevalence of infertility among the Uygur and Kazak women were 26.5％ and 21.7％ respectively,the difference was not statistically significant (P＞0.05).The prevalence of primary infertility among the Uygur and Kazak women were 14.7％,and 8.7％,respectively,the difference was statistically significant (P＜ 0.05).The prevalence of secondary infertility among the Uygur and Kazak women were 11.8％ and 13.0％,respectively,the difference was not statistically significant (P＞0.05).The prevalence of infertility in the Uygur women was correlated with household income,pelvic inflammation,endometriosis and BMI,while the prevalence of infertility in the Kazak women was correlated with age of marriage,endometriosis and the history of ectopic pregnancy.Conclusion The prevalence of infertility was high among the Uygur and Kazak women at reproductive age in Xinjiang.The influencing factors varied with ethnic group.It is necessary to conduct targeted health education and provide early diagnosis and effective treatment.
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Objective To analyze the relationship between the levels of plasma total homocysteine (Hcy) and umbilical arterial with color doppler ultrasound with the severity of preeclampsia(PE).Methods A total of 70 cases of the third trimester of normal pregnancy, 72 cases of mild PE, and 66 cases of severe PE were selected.Then plasma Hcy levels and umbilical arterial with color doppler ultrasound were detected.Results Hcy levels were statistically significant difference among the normal control group,mild PE group,and severe PE group((8.950±0.585), (11.116±0.615), (14.648±0.620) μmol/L,P＜0.05).Umbilical artery blood flow parameter S/D values in severe PE group was significantly higher in mild PE group and in contrg.1 group (3.43 ±0.72 vs.2.86 ± 0.82 vs 2.75 ± 0.56, P＜ 0.05), while the control group, mild PE group had no significant difference(P＞0.05).Resistance index(RI) ,Pulsatility index(PI) in mild PE group and severe PE group were significantly higher than control group(0.60±0.05,0.78±0.07 vs.0.57±0.06;1.24±0.21,1.47±0.64 vs.0.67±0.35), and the differences among three groups were significant(P＜0.05), while the mild PE, severe PE group had no significant difference(P＞0.05).Hcy of mild PE group was positively correlated with S/D, RI and PI(r=0.548,P=0.009;r=0.587,P=0.008;r=0.324,P=0.02).Hcy of severe PE group was positively correlated with S/D,RI and PI(r=0.752,P=0.001;r=0.627,P=0.003;r =0.438,P=0.024).Hcy and the severity of PE was positively correlated (r =0.450, P ＜ 0.05), S/D and the severity of PE was positively correlated(r=0.316, P =0.002).RI, PI and the severity of PE was no correlation (r =0.024, P =0.726;r =0.054,P =0.649).Conclusion The levels of Hcy and S/D were related to the severity of PE.To reduce Hcy,and monitor S/D were new ways for the diagnosis and treatment of pre-eclampsia.
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Objective To investigate the clinical characteristics and differences among Uygur and Han patients with simple severe preeclampsia and their first-degree relatives. Methods To analysis the 100 cases with simple severe preeclampsia among Uygur and Han patients and their first-degree relatives. Results The age and gestational age between Uygur and Han group, there was no significant difference (P>0. 05). Average production time, women with preeclampsia history and BMI of Uygur group were higher than the Han group(P 0. 05). Conclusion During pregnancy, the Uygur should pay more attention to improve diet structure, control body weight, and strengthen their pregnancy health care, so that the disease can be early diagnosed and treated, reducing or avoiding therapeutic preterm labor.
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Objective To investigate the lentivirus-mediated RNA interference(RNAi) reduced human epidermal growth factor 2 (HER2) gene expression in ovarian cancer chemosensitivity.Methods Specifically targeted short hairpin RNA HER2 gene ( shRNA) lentiviral expression vector and negative control sequences lentiviral vector were constructed.stable expression of HER2-shRNA in SKOV3 cell line ( KD) and negative control cell line (NC) were established.The uninfected SKOV3cell lineserved as blank control(CON)group, sensitivity to changes in three kinds of cells to cisplatin chemotherapy in the HER2 gene expression was detected by RT-PCR and Western blot technique, CCK-8 assay CON and KD cells strains were established xenograft model in nude mice in each group to detect the expression of the HER2 protein, as well as by cisplatin chemotherapy tumor size. Results Expression results showed that a stable cell line HER2-shRNA and negative control cell lines had been successfully establish by fluorescence and flow cytometry.RT-PCR results showed KD cells relative HER2 mRNA expression levels compared with the NC group, the difference was statistically significant (P<0.01), whereas no significant difference when compared with the CON group, HER2 mRNA KD Group inhibition rate was (95.5 ± 0.12)%.After transduction specific HER2-shRNA ovarian cancer tumor cells, HER2 protein expression was significantly reduced, growth had slowed down significantly after cisplatin treatment ( P<0.01 ) .Conclusion Lentivirus-mediated HER2-shRNA can effectively inhibit the expression of the HER2 gene, increase chemosensitivity to cisplatin in ovarian cancer.
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This paper probed into the talent training mode of two circles training,alternation of learning and training of bio-pharmaceutical technology through the reform of professional cognitive practice,professional skill training and post practice,etc.The practice teaching reforms carried out above intensify students' professional competence and professional quality.
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ObjectiveTo observe the treatment effects in 48 cases of advanced lung cancer patients,with the immune therapy of the dendritic cells loading of tumor autologous antigen (DCTAA) combining with the cells induced factor of the killer cells(CIK)from the matched umbilical cord blood cells.MethodsThe peripheral blood mononuclear cell(PBMC)from the matched umbilical cord blood cells was seperated,and induced to CIK and DC with some cytokines in vitro, such as CD3McAb, IL-2, IFN-γ IL-1α, etc. After 12 to 15 days, the amplified CIK cells obtained were obtained, with the strict quality control, infused the CIK cells to the patients body back in six times,about(5-8)×109 CIK cells in each time.In the fifth day of the cultivation,DETAA cells were loaded and DCTAA cells were collected in the eighth day,and then hypodermic injection was done. The patient' s general situation after the immune treatment was observed, such as the size of the tumors, clinical symptom score, the quality of life and immune indexes. Karnofsky score, weight, toxic side effects and the patient's survival were also studied.ResultsIn the 48 cases with the DCTAA-CIK treatment, complete remission (CR), partial remission (PR)was 37 cases, the overall remission rate was 77.1％. The improvement rate of clinical symptom scores was from 78.9 ％ to 84.7 ％, the increasing rate of Karnofsky score was 89.6 ％ (43/48). 1-year survival reached to 80.6 ％. There were significant difference in little toxic side effects(P ＜ 0.01). The proportion of CD3, CD4 and NK cells in peripheral blood cells increased significantly (P ＜ 0.01) after DCTAA-CIK cells treatment[(42.21±6.12)％, (24.42±3.01)％, 0.99±0.34, (24.98±3.02) ％; (71.58±7.64) ％, (37.25±2.13) ％, 1.62±0.45, (35.23±4.11) ％](t = 6.34, 5.67, 0.25, 4.43, P ＜0.01).ConclusionThe DCTAA-CIK immune therapy is benefit for advanced lung cancer,not only improve the immune function but also ameliorate the clinical symptoms.
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Objective To analyze and evaluate the impact of trusteeship on hospital management and operating status. Methods The state of affairs, technical merit and financial situation and other datawere invested and analyzed in some trust hospital in Xinjiang from 2007 to 2010. Results The number of outpatients, inpatients and surgeries compared before hosting increased by 30. 3%, 61.3% and 26. 3%, respectively. The average in hospital shortened 1.1 days. The growing medical expenditure of average outpatientand inpatient was controlled at a reasonable range. 33 new technologies were carried out after trusteeship. The total revenue in 2010 increased 54.2% compared to 2007, with the balance between revenue and expenditure. Conclusion Hospital trusteeship helped improve hospital operation and management. Measures for hospital trusteeship should be suited to local conditions and sensible.