Study of photo-ionization and thermal effects on the interaction of a nanosecond extreme ultraviolet laser with copper.

The damage mechanism of a nanosecond extreme ultraviolet (EUV) laser with solid targets is complex and involves thermal and nonthermal effects. In this study, the interaction process of a nanosecond 46.9 nm laser with copper was investigated. A Faraday cup was used to measure the electron signals induced by the laser irradiation. The photo-ionization and thermal effects in the interaction process are discussed according to the results.

Clinical significance of blood lipids, lipoproteins, PTX-3, TTF-1, NSE, and CYFRA21-1 levels in patients with advanced gastric cancer / 中国医师杂志

Objective:To investigate the significance of blood lipids [triglyceride (TG), total cholesterol (TC)], lipoproteins [high-density lipoprotein (HDL), low-density lipoprotein (LDL)], and serum levels of pentraxin-3 (PTX-3), thyroid transcription factor-1 (TTF-1), neuron specific enolase (NSE), and human cytokeratin 21-1 fragment (CYFRA21-1) in patients with advanced gastric cancer, and to provide a basis for the early, middle, and late diagnosis and treatment of gastric cancer.Methods:127 gastric cancer patients admitted to 3201 Hospital from January 2019 to January 2022 were selected as the research subjects. They were divided into early stage group ( n=45), mild stage group ( n=43), and late stage group ( n=39) based on their condition. Enzyme linked immunosorbent assay (ELISA) was used to detect blood lipids (TG, TC), PTX-3, TTF-1, NSE, CYFRA21-1, and chemical precipitation method was used to detect lipoprotein metabolism (HDL, LDL) in the three groups of patients. The differences in blood lipids, lipoproteins, PTX-3, TTF-1, NSE, and CYFRA21-1 between three groups of gastric cancer patients and the late stage group of gastric cancer patients before and after surgery were analyzed. Logistic regression analysis was conducted to investigate the correlation between blood lipids (TG, TC), lipoprotein (HDL, LDL), PTX-3, TTF-1, NSE, CYFRA21-1, and gastric cancer incidence. The predictive value of individual and combined detection of the above indicators for gastric cancer was analyzed using the receiver operating characteristic (ROC) curve analysis. Results:The results showed that the TG, TC, and LDL levels in the late stage group were higher than those in the mild stage and early stage groups (all P<0.05), while the HDL levels were lower than those in the mild stage and early stage groups (all P<0.05). The serum levels of PTX-3, TTF-1, NSE, and CYFRA21-1 were higher than those in the mild stage and early stage groups (all P<0.05). The postoperative levels of TG, PTX-3, TTF-1, NSE, CYFRA21-1, TC, and LDL in the late stage group were significantly lower than those before surgery (all P<0.05) and the HDL level was higher than that before surgery ( P<0.05). The levels of TG, TC, HDL, LDL, PTX-3, TTF-1, NSE, and CYFRA21-1 were correlated with the late onset of gastric cancer (all P<0.05). The ROC curve showed that the area under the ROC curve (AUC) of PTX-3, TTF-1, NSE, and CYFRA21-1 combined detection was significantly higher than that of PTX3, TTF1, NSE, and CYFRA211 alone. Among them, PTX-3+ TTF-1+ NSE+ CYFRA21-1 combined detection had the highest AUC, sensitivity, and specificity. Conclusions:Patients with advanced gastric cancer have abnormal levels of blood lipids (TG, TC), lipoprotein (HDL, LDL), and serum PTX-3, TTF-1, NSE, and CYFRA21-1. Effective intervention measures need to be developed based on the above indicators to improve survival rate.

Clinical study of endoscopic surgery through thoraco mammary approach in treatment of thyroid papillary carcinoma / 中华内分泌外科杂志

Objective:To investigate the effect of endoscopic thyroidectomy through breast milk approach in patients with papillary thyroid carcinoma and its influence on Wnt and integrin signaling pathways.Methods:A total of 136 patients diagnosed with papillary thyroid carcinoma in our hospital from Jul. 2018 to Mar. 2020 were selected and were hospitalized for surgical treatment. According to different surgical procedures, they were divided into a study group (68 cases) and a control group (68 cases) . The control group was treated with open thyroidectomy and the study group was treated with thoracoscopic thyroidectomy. The two groups were compared in terms of immune function [CD4+, CD8+ and CD4+/CD8+], and pain index [PGE2, IL-6, Cor and VAS score]. RT-PCR method was used to detect WNT1, β-catenin, GSK3β and integrin Signal pathway before and after surgery.Results:Three days after operation, compared with the control group, the study group had significantly higher CD4+ and CD4+/CD8+ levels [ (27.62±2.52) vs (24.63±2.67) , (0.66±0.18) vs (0.52±0.13) ], while the CD8+ level was significantly lower [ (41.62±3.54) vs (45.62±3.63) ] ( P<0.001) ; PGE2, IL-6, Cor, VAS of the study group were significantly lower than the control group [ (48.54±9.86) vs (57.21±8.12) , (5.13±0.71) vs (6.99±0.95) , (511.23±67.52) vs (633.12±71.47) , (1.26±0.56) vs (3.99±2.06) ] ( P<0.001) ; WNT1, β-catenin, GSK3β, integrin β1, FAK, Ras, and MAPK mRNA expression levels in the study group were significantly lower than those of the control group[ (1.79±0.15) vs (2.85±0.25) , (1.94±0.15) vs (2.64±0.24) , (2.13±0.19) vs (2.97±0.28) , (1.95±0.17) vs (2.58± 0.23) , (2.15±0.16) vs (2.87±0.22) , (1.95±0.18) vs (2.91±0.27) , (1.89±0.12) vs (2.87±0.31) ] ( P<0.001) . Conclusion:Endoscopic thyroidectomy through thoracolumbar approach can effectively reduce postoperative pain in patients with papillary thyroid cancer, have a smaller impact on immune function, and block the expression of Wnt and integrin signaling pathways to reduce tumor metastasis risk.

Prognostic value of serum P-cadherin level in patients with non-small cell lung cancer / 国际肿瘤学杂志

Objective To investigate the prognostic value of serum P-cadherin( P-cad)level in patients with non-small cell lung cancer(NSCLC). Methods A total of 80 patients with NSCLC in Hanzhong 3201 Hospital of Shaanxi Province from January 2012 to December 2013 were selected as study subjects. The relationships between serum P-cad level and clinicopathological characteristics were analyzed. The Cox regres-sion analysis was used to analyze the risk factors affecting prognosis of NSCLC patients. The survival curve was drawn by Kaplan-Meier method and the difference test was carried out by log-rank method. Results There were significant differences in lymph node metastasis(χ2 = 14. 31,P < 0. 001),vascular invasion(χ2 = 5. 56, P = 0. 018)among NSCLC patients with different levels of serum P-cad. Multivariate Cox regression analysis showed that lymph node metastasis(HR = 0. 856,95％ CI:0. 702-0. 955,P = 0. 012),TNM stage(ⅢA HR =1. 315,95％ CI:1. 058-1. 991,P = 0. 024;ⅢB HR = 1. 448,95％ CI:1. 124-2. 215,P = 0. 011;Ⅳ HR =1. 569,95％ CI:1. 182-2. 441,P < 0. 001)and high level of serum P-cad(HR = 1. 815,95％ CI:1. 224-3. 562,P < 0. 001)were risk factors for progression-free survival in NSCLC patients,and lymph node metasta-sis(HR = 0. 755,95％ CI:0. 652-0. 915,P = 0. 022),poor differentiation(HR = 1. 622,95％ CI:1. 112-2. 015,P < 0. 001),TNM stage(ⅢA HR = 1. 335,95％ CI:1. 064-2. 014,P = 0. 011;ⅢB HR = 1. 489, 95％ CI:1. 129-2. 297,P < 0. 001;Ⅳ HR = 1. 622,95％ CI:1. 192-2. 501,P < 0. 001)and high level of serum P-cad(HR = 1. 677,95％ CI:1. 193-2. 668,P < 0. 001)were risk factors for overall survival of NSCLC patients. The results of Kaplan-Meier survival curve showed that the overall survival and progression-free survival of patients with high level of serum P-cad were shorter than those of patients with low level of serum P-cad(χ2 = 5. 18,P = 0. 015;χ2 = 5. 48,P = 0. 011). Conclusion High level of serum P-cad is closely related to poor prognosis in NSCLC patients.

Relationship between serum and glucocorticoid-regulated kinase 1 expression and prognosis in patients with non-small cell lung cancer / 国际外科学杂志

Objective To investingate the effect of SGK1 expression level on the prognosis of patients with NSCLC,and provide new biological predictors for the prognosis assessment of patients with NSCLS.Methods One hundred and twenty patients with NSCLC received radical resection in Hanzhong 3201 hospital from Jan 2011 to Dec 2013 were selected.There were 75 males and 45 females,age (63.15 ± 16.44) years,age range 45-80 years.According to the results of immunohistochemical staining,the SGK1 cut-off value determined by the integral was determined,and NSCLC patients were divided into SGK1 high expression group (n =70) and SGK1 low expression group(n =50).The relationship between the expression of SGK1 and clinicopathological features (age,sex,smoking history,alcoholism history,BMI,tissue type,tumor diameter,T stage,N stage,TNM stage,differentiation degree) in NSCLC were analyzed,and the overall survival rate in NSCLC were also analyzed.Followup was carried out by telephone or patient admission.The follow-up period was up to June 1,2018.Chest X-ray and ultrasonography were reviewed every 3 to 6 months after operation,and enhanced CT or MRI were performed if the results were abnormal.The measurement data conforming to normal distribution were expressed by t test and showed by (Mean ± SD);the counting data were tested by x2 test;the 5-year overall survival rate was used as the endpoint event for univariate analysis,and the significant variables for univariate analysis were analyzed by COX risk ratio model for multivariate analysis.The cumulative survival curve was drawn by Kaplan-Meier method,and the difference was tested by Log-rank method.Results The expression level of SGK1 in tissues was not related to age,sex,smoking history,alcoholism,BMI,tissue type and tumor diameter (P ＞ 0.05),but it was related to T stage,N stage,TNM stage and differentiation degree (P ＜ 0.05).The univariate and multivariate COX risk ratio model showed that TNM stage and SGK1 expression were independent factors affecting the 5-year overall survival rate of NSCLC patients (P ＜ 0.05).The results of Kaplan-Meier survival curve showed that the 5-year overall survival rate in NSCLC with low expression of SGK1 was significantly higher than that in NSCLC with high expression of SGK1 (P ＜ 0.05).Conclusions The expression of SGK1 in tissues is closely related to the prognosis of patients with NSCLC.The high expression of SGK1 in tissues is not conducive to the prognosis of patients with NSCLC.

Effect of CT guided implantation of 125Ⅰ radioactive particles in superficial malignant tumors / 实用放射学杂志

Objective To investigate the short-term clinical effect of CT guided 125Ⅰ radioactive particle therapy in superficial malignant tumor. Methods The clinic data of 28 patients with metastatic superficial malignant tumor in our hospital were analyzed retrospectively.All patients were treated with CT guided 125Ⅰ radioactive particle therapy.The short-term effects,1 year survival rate and 1 year progression free survival rate of the patients were compared.Results Objective remission rate(ORR)and disease control rate(DCR)after 6 months were 92.86% and 100.00%.1 year overall survival and 1 year progression free survival were 96.43%(27/28)and 82.14%(23/28), respectively.The median overall survival and median progression free survival were 26.978 months (95%CI:22.558-31.399)and 16.932 months (95 % CI:14 .471-19.393).There were 27 cases of 0-Ⅱ degree adverse reactions,1 case of grade Ⅲ adverse reactions and no grade Ⅳ adverse reactions.No signs of 125Ⅰ radioactive particle translocation,vascular embolism and vascular rupture were found. Conclusion 125Ⅰ radioactive particle treatment of superficial malignant tumor has a definite short-term curative effect,with overall survival and progression free survival longer and higher safety,which can be considered in clinical application.

Clinical Effect of Ubenimex Capsules Combined with SOX Chemotherapy on Treatment of Advanced Gastric Cancer / 现代生物医学进展

Objective:To study the clinical effect and safety of ubenimex capsules and SOX chemotherapy on the advanced gastric cancer.Methods:90 patients with advanced gastric cancer who were treated in our hospital from September 2013 to September 2015 were selected and randomly divided into the observation group (n=45) and the control group (n=45).The patients in the control group were treated with SOX chemotherapy,while the patients in the observation group were treated with ubenimex capsules on the basis of control group.Then the serum levels of MMP-2 and MMP-9,the immune functions,the clinical efficacy,the adverse reactions and survival rate of two groups were observed and compared before and after the treatment.Results:After treatment,the CD4+,CD4+/CD8+ in the observation group were higher than those of the control group (P＜0.05);The levels of MMP2 and MMP-9 in the observation group were lower than those of the control group (P＜0.05);The total effective rate of the observation group was higher than that of the control group [68.89％(31/45) vs 48.89％(22/45)] (P＜0.05);The incidence of thrombocytopenia,leukopenia,nausea and vomiting and abnormal liver functions in the observation group was lower than that of the control group (P＜0.05);The survival rate of the observation group was higher than that of the control group at 6 months and 12 months [93.33％ (42/45) vs 77.78％ (35/45),82.22％ (37/45) vs 57.78％ (26/45)](P＜0.05).Conclusion:Compared with SOX chemotherapy alone,ubenimex capsules and SOX chemotherapy could effectively improve the immune function,enhance the long-term survival rate with high safety of patients with advanced gastric cancer.

Research on LPA and Hippo-YAP signaling pathway in promoting invasion and metastasis of TNBC cell MDA-MB-231 / 医学研究生学报

Objective Triple negative breast cancer(TNBC), a special breast cancer subtype, is lack of effective target therapy.The article aimed to investigate the role of lysophosphatidic acid (LPA) and Hippo Yes-associated protein (Hippo-YAP) signaling pathway in TNBC invasion and metastasis and the mechanisms.Methods The specific small interfering RNA (siRNA) of YAP was synthetized in vitro, and was transfected into MDA-MB-231 cells using liposome transfection.The experiment was divided into YAP-siRNA group, positive control group and blank control group.Each group is provided with 2 parallel holes.Evaluation was made on the effects of each group on Hippo-YAP, the mechanisms and regulation on upstream and downstream molecules of Hippo-YAP pathway.Results In experiment group, YAP content, the capacity of invasion and metastasis after transfection ([0.035±0.005], [2.200±1.000], [3.500±0.800]) significantly decreased compared with positive control group([0.343±0.012], [27.600±5.100], [22.300±5.000]) and blank control group([0.384±0.017], [26.500±4.800], [22.350±6.000]) (P<0.05).YAP expression levels at 60 min, 120 min, and 240 min in experiment group significantly decreased compared with positive control group and blank control group (P<0.05).YAP relative expression levels of 10, 20, 50 μmol/Lwere significantly lower than those of positive control group and blank control group (P<0.05).After respective interference of C3 transferase and Y27623, significant difference was found in the pYAP mRNA contents of experiment group([0.255±0.052], [0.326±0.017]), blank control group([0.048±0.032], [0.534±0.017]) and positive control group([0.052±0.021], [0.528±0.024])(P<0.05).The expression levels of YAP mNA and AREG mNA significantly increased in experiment group([0.176±0.032], [0.263±0.008]) compared with blank control group([0.043±0.013], [0.263±0.008]) and positive control group([0.049±0.025], [0.057±0.043])(P<0.05).Conclusion LPA induces breast cancer invasion and metastasis, which is YAP-dependent, time-dependent and concentration-dependent.LPA-Hippo-YAP singaling pathway may be one of the mechanisms promoting delayed metastasis of TNBC.

Expression of lysophosphatidic acid receptors in breast cancer and their significances / 肿瘤研究与临床

Objective To evaluate the differential expression of lysophosphatidic acid receptor (LPAR) in breast cancer (BC), and its relationship with clinicopathological features of BC patients. Methods The qRT-PCR and immunohistochemical staining were used to detect the LPAR expression in 37 normal tissues, 55 benign disease tissues and 82 BC tissues, besides, the correlation of LPAR expression with clinicopathological data was also analyzed. Results The expression levels of LPAR2 and LPAR3 mRNA and protein in BC tissues were higher than those in normal benign tissues (all P0.05). LPAR1 expression was not associated with clinicopathological features in BC tissues (P>0.05). LPAR2 expression in postmenopausal patients was higher than that in premenopausal patients (χ2=4.821, P<0.05). LPAR3 expression was significantly associated with nodal metastasis, estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2) in BC tissues (all P<0.05). Conclusion LPAR in BC tissues has differential expression, which is associated with nodal metastasis, ER, PR and HER2.

Oxidative stress response in diabetic nephropathy rats following injection of embryonic stem cells via the tail vein / 中国组织工程研究

BACKGROUND:Occurrence and development of diabetic complications is closely related to the severity of oxidative stress imbalance in the body. OBJECTIVE:To investigate the effect of embryonic stem cel s on oxidative stress response of rats with diabetic nephropathy. METHODS:Primarily cultured rat embryonic stem cel s were observed for cel morphology and surface antigen detection. Sprague-Dawley rats were divided into experimental group (two injections of embryonic stem cel s via the tail vein), model group (injection of the same volume of PBS), and normal control group (with no modeling, intraperitoneal injection of sodium citrate-citrate buffer). In the former two groups, the rats were intraperitoneal y injected sodium citrate-citrate buffer diluted streptozotocin to establish diabetic nephropathy models before treatment. At 5 weeks after the last injection, blood glucose level, renal function indicators (urine protein/urine creatinine, blood urea nitrogen and serum creatinine) were tested in each group;contents of malondialdehyde and protein carbonyl were detected in the kidney;the expression level of superoxide dismutase was detected by western blot assay. RESULTS AND CONCLUSION:The embryonic stem cel s were oval or round, with clear boundary and good refraction, and highly expressed Oct-4 and SSEA-1. Compared with the control group, renal biochemical indicators, malondialdehyde and protein carbonyl contents were significantly increased, while the expression level of superoxide dismutase was decreased dramatical y in the model group and experimental group (P<0.05);compared with the model group, the renal biochemical indicators, malondialdehyde and protein carbonyl contents were dropped significantly in the experimental group, but the expression of superoxide dismutase was significantly rebounded (P<0.05). Taken together, embryonic stem cel s can reverse the occurrence and development of diabetic nephropathy by inhibiting oxidative stress in progress.

Mouse embryonic stem cell transplantation for treatment of osteoporosis / 中国组织工程研究

BACKGROUND:Because of poor therapeutic effects of drug treatment, stem cel transplantation has become a hot spot in the treatment of osteoporosis. OBJECTIVE:To investigate the therapeutic efficacy of embryonic stem cel transplantation in the treatment of estrogen deficiency-induced osteoporosis. METHODS: Thirty female healthy C57BL/6 mice were randomly divided into model, sham-operated, and treatment groups, with 10 mice in each group. Ovariectomized models were established. In the treatment group, embryonic stem cels were injected by tail vein at 1 day after modeling, and there was no treatment in the other two groups. After 10 hours of treatment, flow cytometry was used to detect the apoptosis of T-lymphocytes; after 3 days of treatment, ELISA assay was used to detect the level of tumor necrosis factor-α in mouse serum; after 1 month of treatment, micro-CT was employed to determine the number of bone trabeculae and bone mineral density. RESULTS AND CONCLUSION: Compared with the sham-operated group, the number of bone trabeculae and bone mineral density were lower in the model group, while the serum level of tumor necrosis factor-α was higher in the model group. After embryonic stem cel transplantationvia the tail vein, the number of bone trabeculae and the bone volume fraction were increased certainly, the serum level of umor necrosis factor-α was decreased, and the number of apoptotic T-lymphocytes was increased in the treatment group, which significantly differed from those in the model group (P< 0.05). These findings indicate that embryonic stem cel therapy has a certain effect on the treatment of osteoporosis, which may play a role through immune regulation.

Hydrogen peroxide influences embryonic stem cell transplantation in the treatment of myocardial infarction / 中国组织工程研究

BACKGROUND:Specific mechanism of stem cel transplantation for treatment of myocardial infarction has been not very clear. OBJECTIVE:To investigate the effect of H2O2 METHODS: Twenty-four C57BL/6 mice, 9 weeks old, were randomized into myocardial infarction group, embryonic stem cels group, H-pretreated mouse embryonic stem cels in vitro for treatment of myocardial infarction. 2O2 pretreatment group, with eight mice in each group, which were respectively injected 150 μL PBS, 150 μL embryonic stem cels (1×106), and 150 μL H2O2-pretreated embryonic stem cels (1×106 RESULTS AND CONCLUSION:Compared with the myocardial infarction group, the left ventricular end-diastolic diameter and left ventricular end-systolic inner diameter were significantly decreased in the embryonic stem cels ) at 1 week after modeling. At 3 weeks after transplantation, left ventricular end-diastolic diameter, left ventricular end-systolic inner diameter and left ventricular ejection fraction of mice in each group were measured by ultrasound. Myocardial colagen content was detected by Sirius red staining and the average colagen volume fraction was calculated. group and H2O2 pretreatment group, while the left ventricular ejection fraction was significantly increased (P < 0.05). Heart weight/body weight, left ventricular weight/body weight, and average colagen volume fraction were significantly lower in the embryonic stem cels group and H2O2 pretreatment group than the myocardial infarction group (P < 0.05). Therefore,in vitro H2O2 pretreatment enhances reduce the post-myocardial infarction myocardial fibrosis and greatly improve heart function.

Hypoxia-inducible factor-1alpha gene-modified umbilical cord blood mesenchymal stem cells transplantation in the treatment of spinal cord ischemia-reperfusion injury / 中国组织工程研究

BACKGROUND:There are no available therapies for spinal cord ischemia-reperfusion injury, and stem cel transplantation is a focused topics. OBJECTIVE:To observe the therapeutic effect of hypoxia-inducible facotr-1α gene-modified umbilical cord blood mesenchymal stem cels (UC-MSCs) transplantedvia the infrarenal abdominal aorta on spinal cord ischemia-reperfusion injury in rats. METHODS:Thirty adult female Sprague-Dawley rats were randomly divided into three groups, with 10 rats in each group. The infrarenal abdominal aorta of rats was occluded surgicaly for 1 hour, and then the spinal cord reperfusion was restored. At 2 hours after reperfusion, 1 mL of 10% PBS, UC-MSCs suspension and hypoxia-inducible factor-1α-modified UC-MSCs suspension was injectedvia the infrarenal abdominal aorta, respectively, in the three groups. At 1, 6, 12 days after injection, Basso-Beattie-Bresnahan scores were recorded and western blot assay was used to detect hypoxia-inducible factor-1α protein expression in the spinal cord. The motor-evoked potential was determined at 12 days after injection. RESULTS AND CONCLUSION: Compared with the control group, the Basso-Beattie-Bresnahan scores were significantly higher (P < 0.05), the expression of hypoxia-inducible factor-1α protein in the spinal cord tissue was significantly increased (P < 0.05), the motor-evoked potential latency was shortened (P < 0.05) and the amplitude was increased (P< 0 .05) in the untransfected group and transfection group. Compared with the untransfected group, the Basso-Beattie-Bresnahan scores were significantly higher (P < 0.05), the expression of hypoxia-inducible factor-1α protein in the spinal cord tissue was significantly increased (P < 0.05), the motor-evoked potential latency was shortened (P < 0.05) and the amplitude was increased (P < 0 .05) in the transfection group. Above al, umbilical cord blood mesenchymal stem cel transplantation modified by hypoxia-inducible factor 1α has better effects on spinal cord ischemia-reperfusion injury.

Dopamine content in the stratum of Parkinson’s disease rats after transplantation of tyrosine hydroxylase-modified human umbilical cord blood mesenchymal stem cells / 中国组织工程研究

BACKGROUND:Stem cel therapy is superior to drug therapy for recovery of patient’s physiological mode, and cel transplantation therapy is becoming a trend. OBJECTIVE:To observe the changes in dopamine content in the stratum of Parkinson’s disease rats after transplantation of tyrosine hydroxylase-modified human umbilical cord blood mesenchymal stem cel s. METHODS:After identification by enzyme digestion, pEGFP-C2-TH plasmid was transfected into the fourth generation of human umbilical cord blood mesenchymal stem cel s by electroporation method, and then transfected cel s were injected into the right cerebral ventricle of Parkinson’s disease rats (experimental group). PBS injection was performed in the control group. Migration of dopamine in the brain tissue of rats was observed, and the content of dopamine was detected by high performance liquid chromatography. RESULTS AND CONCLUSION:At 8 weeks after cel transplantation, the cel s gradual y migrated to the ventricles;after 12 weeks, the cel s migrated to the cortex, and expressed tyrosine hydroxylase antigen. Meanwhile, the content of dopamine was significantly higher in the experimental group than the control group (P<0.05). These results reveal that the intraventricular transplantation of tyrosine hydroxylase-modified human umbilical cord blood mesenchymal stem cel s has obvious therapeutic effect on Parkinson’s disease rats.

Secretion and expression of vascular endothelial growth factor and interleukin-8 by SH-SY5Y human neuroblastoma cells / 生物工程学报

To establish vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8) as secretary biomarkers for cell growth on topographic substrates, we have evaluated the secretion and expression of these 2 factors by SH-SY5Y human neuroblastoma cells on poly-L-lactide (PLLA) micropillar arrayed topographic substrates. We fabricated topographic substrates with UV lithography, silicon etching and polydimethylsiloxane-based replica molding, and interfaced SH-SY5Y human neuroblastoma cells with both the topographic substrates and PLLA flat substrates. Cell morphology and spreading were examined with scanning electron microscopy. The secretion and mRNA expression of VEGF and IL-8 were evaluated with enzyme linked immunosorbent assay (ELISA) and real time qPCR, respectively, 24 hours after cell plating. We successfully achieved 4 topographic substrates with a nominal pillar diameter of 2 microm and 4 microm, and a nominal pillar spacing of 2 microm and 7 microm. We found that the secretion and mRNA expression of VEGF and/or IL-8 by SH-SY5Y cells on 2-2 microm (pillar diameter-spacing), 4-2 microm and 4-7 microm topographic substrates were upregulated in comparison to those by cells on PLLA flat substrate, 24 hours after cell plating. Furthermore, both cytokines were even more substantially upregulated on the 2-7 microm substrate than on the other 3 topographic substrates. Compared to those on PLLA flat substrate, cells on topographic substrates showed significant changes in morphology (spreading area, perimeter and roundness), and the increase in the secretion and mRNA expression of VEGF and IL-8 was accompanied with a decrease in cell spreading areas. These results provided evidence that pillar arrayed topography was an important microenvironmental factor in affecting VEGF and IL-8 expression or secretion, and VEGF and IL-8 might serve as important secretary biomarkers for growth on topographic substrates by SH-SY5Y cells.

Effect of shRNA silenced aromatic hydrocarbon receptor on WNT signaling pathway during the differentiation of cardiac myocytes / 临床儿科杂志

Objective To explore the effect of shRNA silenced aromatic hydrocarbon receptor (AHR) on WNT signaling path-way during the differentiation of P19 cells into cardiac myocytes. Methods The eukaryotic expression vector of mouse AHR gene was designed and constructed. The interference plasmid was transfected into P19 cell and the positive stains to AHR gene silencing were screened by G418. The mRNA expression of important genes GSK3βandβ-catenin were evaluated by real-time fluorescent quantita-tive PCR during the differentiation of P19 cells. Results The constructed AHR-shRNA plasmid significantly inhibited the expression of AHR gene. Along with the differentiation of P19 cell into cardiac myocytes, in the interference group the expression ofβ-catenin gene was lower whereas the expression of GSK3βgene was elevated than those of control group with significant differences (all P<0.01). Conclusions The interference of AHR gene expression can regulate WNT signaling pathway in the development of heart.

Cloning, expression, identification and bioinformatics analysis of Rv3265c gene from Mycobacterium tuberculosis in Escherichia coli

OBJECTIVE@#To clone and express Rv3265c gene of Mycobacterium tuberculosis in Escherichia coli (E. coli) under optimistic conditions, obtain and identify protein expressed, analyze the structure and characteristics of the protein using bioinformatics methods for future applications.@*METHODS@#Rv3265c gene from Mycobacterium tuberculosis H37Rv was amplified by polymerase chain reaction, and was cloned into the pET-30a vector after purification and recovery. The recombinant plasmid was sequenced and expressed in E. coli BL21(DE3), and then purified and identified by western blotting. The essential physical-chemical properties of the protein were predicated by bioinformatics tools, including subcellular location, secondary structure, domains, antigenic epitopes, etc. Tertiary structure of the protein based on homology modeling was established, while multi-sequence homological alignment and phylogenetic analysis were proformed.@*RESULTS@#The recombinant protein was obtained in soluble fraction from expression system in E. coli BL21(DE3) carrying pET30- Rv3265c plasmid, and Rv3265c gene was expressed correctly. Bioinformatics analysis showed the protein contained no signal peptide and transmembrane helices, located outside of membrane. Secondary structure analysis revealed it contained α-helix, extended strand and random coil, 46.8%, 14.6%, 38.6%, respectively. Furthermore, it possessed six potential antigenic epitopes, one glycosyl transferase domain. A simple three-dimensional model of this protein was constructed by Swiss-model sever. Both sequences and structures were conservative and especial either in gene or in protein.@*CONCLUSIONS@#Rv3265c gene might be a desirable molecular target for anti-tuberculosis drug and vaccine. The purified protein from expression will be utilized to study the kinetics of L-rhamnosyltransferase and to develope an enzyme assay for screening vaccine or drug.

Bioinformatics analysis for structure and function of CPR of Plasmodium falciparum

OBJECTIVE@#To analyse the structure and function of NADPH-cytochrome p450 reductase (CYPOR or CPR) from Plasmodium falciparum (Pf), and to predict its' drug target and vaccine target.@*METHODS@#The structure, function, drug target and vaccine target of CPR from Plasmodium falciparum were analyzed and predicted by bioinformatics methods.@*RESULTS@#PfCPR, which was older CPR, had close relationship with the CPR from other Plasmodium species, but it was distant from its hosts, such as Homo sapiens and Anopheles. PfCPR was located in the cellular nucleus of Plasmodium falciparum. 335aa-352aa and 591aa - 608aa were inserted the interior side of the nuclear membrane, while 151aa-265aa was located in the nucleolus organizer regions. PfCPR had 40 function sites and 44 protein-protein binding sites in amino acid sequence. The teriary structure of 1aa-700aa was forcep-shaped with wings. 15 segments of PfCPR had no homology with Homo sapien CPR and most were exposed on the surface of the protein. These segments had 25 protein-protein binding sites. While 13 other segments all possessed function sites.@*CONCLUSIONS@#The evolution or genesis of Plasmodium falciparum is earlier than those of Homo sapiens. PfCPR is a possible resistance site of antimalarial drug and may involve immune evasion, which is associated with parasite of sporozoite in hepatocytes. PfCPR is unsuitable as vaccine target, but it has at least 13 ideal drug targets.

Bioinformatics analysis and prediction for structure and function of nitric oxide synthase and similar proteins from Plasmodium berghei

OBJECTIVE@#To search and analyze nitric oxide synthase (NOS) and similar proteins from Plasmodium berghei(Pb).@*METHODS@#The structure and function of nitric oxide synthase and similar proteins from Plasmodium berghei were analyzed and predicted by bioinformatics.@*RESULTS@#PbNOS were not available, but nicotinamide adenine dinucleotide 2'-phosphate reduced tetrasodium (NADPH)-cytochrome p450 reductase(CPR) were gained. PbCPR was in the nucleus of Plasmodium berghei, while 134aa-229aa domain was localize in nucleolar organizer. The amino acids sequence of PbCPR had the closest genetic relationship with Plasmodium vivax showing a 73% homology. The tertiary structure of PbCPR displayed the forcep-shape with wings, but no wings existed in the tertiary structure of its' host, Mus musculus(Mm). 137aa-200aa, 201aa-218aa, 220aa-230aa, 232aa-248, 269aa-323aa, 478aa-501aa and 592aa-606aa domains of PbCPR showed no homology with MmCPRs', and all domains were exposed on the surface of the protein.@*CONCLUSIONS@#NOS can't be found in Plasmodium berghei and other Plasmodium species. PbCPR may be a possible resistance site of antimalarial drug, and the targets of antimalarial drug and vaccine. It may be also one of the mechanisms of immune evasion. This study on Plasmodium berghei may be more suitable to Plasmodium vivax. And 137aa-200aa, 201aa-218aa, 220aa-230aa, 232aa-248, 269aa-323aa, 478aa-501aa and 592aa-606aa domains of PbCPR are more ideal targets of antimalarial drug and vaccine.

Prokaryotic expression, identification and bioinformatics analysis of fbpB-esxA fusing gene from Mycobacterium tuberculosis

OBJECTIVE@#To obtain fbpB-esxA fusing gene of Mycobacterium tuberculosis (MTB), express the encoded fusing protein in Escherichia coli (E. coli), identify protein acquired, and predict the structure and function of the protein utilizing methods of bioinformatics.@*METHODS@#fbpB and esxA gene were amplified from genome of MTB H37Rv by PCR. The fbpB-esxA fusing gene ligated by (Gly(4)Ser)(3) linker was gained by means of Gene Splicing by Overlapping Extension PCR (SOE-PCR), and fusing gene was cloned into expression vector pET-30a. The recombinant plasmid was sequenced and expressed in E. coli BL21(DE3). The protein was identified by Western blot using anti-HIS antibody. Secondary structure and antigenic epitopes of the protein were predicting using tools of bioinformatics.@*RESULTS@#The DNA sequences of fbpB-esxA were identical with that published by GenBank. The Ag85B-ESAT-6 fusion protein about 50 kDa comprised 485 amino acids was efficiently produced from expression system in E. coli BL21(DE3) under the induction of IPTG. Bioinformatics analysis showed the protein contained one transmembrane region and fourteen potential antigenic epitopes.@*CONCLUSIONS@#The Ag85B-ESAT-6 fusion protein is successfully expressed with N-terminal HIS-tag. Gel filtration demonstrated that it exists as insoluble inclusion bodies mainly. The existence of linker doesn't affect immunogenicity of Ag85B and ESAT-6. It will allow for characterization in vitro and establish a foundation of further function research such as vaccine or diagnostic reagent.