Kang-Ai Injection Inhibits Gastric Cancer Cells Proliferation through IL-6/STAT3 Pathway / 中国结合医学杂志

OBJECTIVE@#To explore the mechanisms underlying the proliferative inhibition of Chinese herbal medicine Kang-Ai injection (KAI) in gastric cancer cells.@*METHODS@#Gastric cancer cell lines MGC803 and BGC823 were treated by 0, 0.3%, 1%, 3% and 10% KAI for 24, 48 and 72 h, respectively. The cell proliferation was evaluated by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. The apoptosis and cell cycle were evaluated by flow cytometry. Interleukin (IL)-6 mRNA and protein expression levels were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and enzyme-linked immune sorbent assay (ELISA), respectively. The protein expression levels of cyclin A, cyclin E, cyclin B1, cyclin D1, p21, retinoblastoma (RB), protein kinase B (AKT), extracellular regulated protein kinases (ERK), signal transducer and activator of transcription (STAT) 1 and STAT3 were detected by Western blot.@*RESULTS@#KAI inhibited the proliferation of MGC803 and BGC823 gastric cancer cells in dose- and time-dependent manner. After treated with KAI for 48 h, the proportion of G1 phase was increased, expression level of cyclin D1 and phosphorylation-RB were down-regulated, whereas the expression of p21 was up-regulated (all P<0.01). Furthermore, 48-h treatment with KAI decreased the phosphorylation level of STAT3, inhibited the mRNA and protein expressions of IL-6 (all P<0.01). IL-6 at dose of 10 ng/mL significantly attenuated the proliferative effect of both 3% and 10% KAI, and recovered KAI-inhibited STAT3 phosphorylation and cyclin D1 expression level (all P<0.01).@*CONCLUSION@#KAI exerted an anti-proliferative function by inhibiting IL-6/STAT3 signaling pathway followed by the induction of G1 phase arrest in gastric cancer cells.

Application of Methylprednisolone Sodium Succinate Combined with Tropisetron in Prevention of Nausea and Vomiting under Microvascular Decompression of Hemifacial Spasm / 中国医学科学院学报

Objective To evaluate the effect of methylprednisolone sodium succinate combined with tropisetron on postoperative nausea and vomiting(PONV)under microvascular decompression of hemifacial spasm.Methods From January to June 2019,485 patients undergoing microvascular decompression for facial spasm at Department of Neurosurgery,Peking University People's Hospital were randomly assigned into two groups with random number table method.For group A(n=242),2 ml saline was administrated by intravenous drip before induction and 5 mg tropisetron after operation.For group B(n=243),40 mg methylprednisolone sodium succinate was administrated by intravenous drip before induction and 5 mg tropisetron after operation.The anesthesia time,operation time,and incidence of PONV in 0-24 h and 24-48 h were recorded for the comparison of the remedial treatment rate of nausea and vomiting between the two groups.Results There was no significant difference in age,gender,smoking history,body mass index value,American Society of Anesthesiologists score,medical history,surgical side,PONV history,operation time or anesthesia time between the two groups(all P > 0.05).The incidence of PONV in group A was 35.5% and 18.2% during 0-24 h and 24-48 h,respectively,which was significantly higher than that(18.5%,χ

Treatment experience of a case of brodifacoum poisoning / 中国职业医学

OBJECTIVE: To analyze the experience of diagnosis and treatment of a case of brodifacoum poisoning. METHODS: The clinical data of a case of unexplained brodifacoum poisoning was retrospectively analyzed. RESULTS: The patient went to the doctor for unexplained bleeding. The bleeding symptoms included nasal o blood ozing, blood in saliva and skin ecchymosis. Blood anticoagulative rodenticide test showed positive with brodifacoum. The results of coagulative function tests showed that the indexes of partial prothrombin time, prothrombin time and fibrinogen were increased. The patient was diagnosed as brodifacoum poisoning based on the clinical symptoms and laboratory test results. The combined use of 40 mg/d of vitamin K_1 and frozen plasma improved the clotting time and quickly alleviated the bleeding symptoms of the patient. However, the patient′s bleeding symptoms recurred when vitamin K_1 was discontinued. The patient was hospitalized for 62 days and then discharged. With follow-up one month after discharge, the patient showed no bleeding symptoms, but brodifacoum could still be detected in the blood. CONCLUSION: The symptoms of brodifacoum poisoning may relapse and the treatment course is long. Vitamin K_1 could be used as the first-choice medicine for the treatment of brodifacoum poisoning, but its usage needs to be optimized.

Tratamiento satisfactorio con fluconazol de la candidemia relacionada con el catéter porCandida haemulonii: informe sobre dos pacientes neonatales / Successful fluconazole treatment of catheter-related candidemia caused by Candida haemulonii: report of 2 neonatal cases

La Candida haemulonii forma parte de la especie Candida no albicans. La candidemia por C. haemulonii es sumamente infrecuente, pero mortal, en los recién nacidos. Se informa sobre los dos primeros recién nacidos con candidemia por C. haemulonii en China tratados con fluconazol y se revisan dos artículos informados con anterioridad. Nuestro informe incrementa la sensibilización sobre la candidemia por C. haemulonii en recién nacidos críticos y resalta la importancia de un diagnóstico y un tratamiento tempranos de esta infección mortal.

Candida haemulonii forms part of the non-albicans Candida species. The candidemia caused by C. haemulonii is extremely rare but fatal in neonates. We reported the first two neonates with C. haemulonii candidemia in China which were treated with fluconazole and reviewed two papers previously reported. Our report adds further awareness on C. haemulonii candidemia in critical neonates and points out the importance of an early diagnosis and treatment of this fatal infection.

Effects of TGF-β1 on the expression and secretion of cytokines induced by Aβ in hippocampal neuron and microglia co-cultures / 中国应用生理学杂志

OBJECTIVE@#To investigate the neuroprotective effects of transforming growth factor beta 1(TGF-β1) on the expression and secretion of cytokines induced by Aβ in hippocampal neurons and microglial co-cultures.@*METHODS@#Hippocampal neurons and microglia obtained from SD rat were co-cultured. TGF-β1 was applied on day 5 after the neurons and microglia co-cultures were incubated at the concentrations of 5 or 20 ng/ml, Aβ was added 1 h following TGF-β1 application at a concentration of 5 μmol/L. They were incubated for 72 h and then assessed for further studies. Western blot analyses were employed to examine the expression of inducible nitric oxide synthase (iNOS); Real-time PCR and ELISA were used to detect the mRNA expression and secretion of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and insulin-like growth factor-1 (IGF-1).@*RESULTS@#In the hippocampal neuron-microglia co-cultures, Aβ induced upregulation of iNOS, TNF-α and IL-1β, downregulation of IGF-1. TGF-β1 pretreatment ameliorated the pro-inflammatory effects caused by Aβ.@*CONCLUSIONS@#TGF-β1 significantly inhibits the increase in inflammatory cytokines and the decrease in neurotrophic factor which are caused by Aβ-induced microglia activation.

Galactomannan test combined with CD4+T lymphocytes for the diagnosis of invasive aspergillosis in patients with AIDS / 中国感染控制杂志

Objective To evaluate the effect of galactomannan(GM) test combined with CD4+ T lymphocyte detection on early diagnosis of invasive aspergillosis (IA) in patients with acquired immunodeficiency syndrome (AIDS).Methods 197 AIDS patients who were suspected with IA in a hospital from January 2014 to December 2016 were analyzed retrospectively,they were divided into confirmed IA group (n =35),clinically diagnosed IA group (n=96,suspected cases),and non-IA group(n =66),sensitivity and specificity of GM test and GM test combined CD4+ T lymphocyte counting for diagnosing IA were compared.Results In confirmed IA group,clinically diagnosed IA group,and non-IA group,the medium values of GM (minimum,maximum) were 1.29(0.65,1.84)pg /mL,0.91(0.36,1.23)pg /mL,and 0.11(0.28,0.72)pg /mL respectively,CD4+ T lymphocyte counting were 45 (29,69)cells/μL,79(35,99) cells/μL,and 89 (59,158) cells/μL respectively,GM value and CD4+ T lymphocyte counting among three groups were significantly different(all P＜0.05).The sensitivity and specificity of single GM test for diagnosing IA in AIDS patients were 64.9％ and 72.7％ respectively;sensitivity and specificity of two consecutive GM test within one week for diagnosing IA were 72.5 ％ and 95.5 ％ respectively;sensitivity and specificity of GM test combined CD4+ T lymphocyte counting were 86.3％ and 90.9％ respectively.Conclusion GM test has better diagnostic value for IA in AIDS patients,continuous GM test and GM test combined CD4+ T lymphocyte counting will further improve the clinical diagnostic value for IA.

The effect of positive pressure ventilation on dust removal in whole lung lavage / 中国职业医学

OBJECTIVE: To explore the effect of positive-pressure ventilation on dust removal in whole lung lavage( WLL).METHODS: By random sample method,21 patients with stage Ⅱ or Ⅲ pneumoconiosis were chosen for different WLL.Using the patients' own left and right lung was used for matched control study. The positive pressure ventilation was performed at the end of the 3rd,6th,9th and 12 th lavage in treatment lung( treatment groups). The positive-pressure ventilation was not implemented at the end of the 3rd and 6th lavage in the contralateral lung( control groups) but implemented at the end of the 9th,11 th and 12 th lavage. The recovery of lavage fluid,dust and dust concentration drained from 4th to 9th lavage were compared in the two groups. RESULTS: There was no statistical difference in the recovery of the lavage fluid in the 4th to 9th lavage in the two groups( P > 0. 05). The amount of dust and the dust concentration in the fourth lavage drainage in the treatment group was higher than that in the control group( P < 0. 01). The amount of dust and the dust concentration in the 6th,8th and 9th lavage drainage in the treatment group was lower than that in control group( P < 0. 01). The amount of dust and the dust concentration in the 3rd positive pressure ventilation were higher than that in the 6th positive pressure ventilation in the treatment group( P < 0. 01). The total amount of dust in the treatment group was higher than that in the the control group( P < 0. 01). CONCLUSION: In whole lung lavage,the positive pressure ventilation can accelerate the discharge of dust in the lung of patients with pneumoconiosis.

Analysis of Risk Factors and Therapeutic Strategies for Relapse of Leukemia after Allogeneic Hematopoietic Stem Cell Transplantation / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To analyze the risk factors of patients with relapsed leukemia after allogeneic hematopoietic stem cell transplantation, and to explore the therapeutic strategies for recurrence.</p><p><b>METHODS</b>The Cox proportional hazard regression model was used for univariate and multivariate analysis of transplantation-related index, a single center retrospective study of clinical data of 202 cases of leukemia received allo-HSCT from March 2004 to October 2014 had been conducted to screen the risk factors for recurrence after transplantation.</p><p><b>RESULTS</b>In the leukemia patients received allo-HSCT, 68 cases relapsed. The relapse rate was 33.6%. The median time of relapse was 4(1.5-26 ) months. Univariate analysis indicated that there were 5 risk factors related with the disease relapse(P<0.05), including the type of disease, extramedullary disease prior to transplant, the course of induced remission, the status of disease at HSCT and chronic graft versus host disease(cGVHD). Multivariate analysis showed that extramedullary disease prior to transplant(RR=2.622, 95%CI 1.139-6.037), the course of induced remission(RR=1.156, 95%CI 0.682-1.957), cGVHD (RR=1.728,95%CI 0.999-2.991) were independent risk factors for relapse of the patients received transplantation. Treatment strategies for the relapsed patients included withdraw immunosuppressant, donor lymphocyte infusion, systemic chemotherapy and local radiotherapy, targeted therapy, and second transplantation. Individualized choice was needed according to the relapsed site. The relapse-related mortality was 25.2%.</p><p><b>CONCLUSION</b>The relapsed patients with leukemia after allo-HSCT have poor prognosis, early interference has good effect. The evaluation and prevention of risk factors before transplantation is even more important.</p>

Effect of Pinggan Qianyang Recipe Containing Serum on Angiotensin II Induced Vascular Smooth Muscle Cell Proliferation and Migration and DNA Methylation / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effect of Pinggan Qianyang Recipe (PQR) on inhibiting angiotensin II (Ang II) induced proliferation and migration of vascular smooth muscle cells (VSMCs) and changes of DNA methylation.</p><p><b>METHODS</b>VSMCs were cultured using tissue explant method, and PQR containing serum was prepared. Primarily cultured VSMCs were divided into four groups, the normal group, the model group, the folate group (folic acid intervention) , and the PQR group. The proliferation and migration of VSMCs was duplicated by Ang II. After 24-h Ang II induced culture, 40 microg/mL folic acid was added to the folate group for 48 h, while 5% PQR containing serum was added to the PQR group for 48 h. The cell growth curve of VSMCs was drawn by using Cell Counting Kit (CCK-8). The proliferative activity of VSMC was determined by MTT assay. The migration of VSMCs was measured by Millicell chamber. The general level of cytosine methylation in cell nucleus was detected via 5-mC antibodies immunofluorescence, and mRNA expression levels of DNA methyltransferase 1 (DNMT1) were measured by Real-time q-polymerase chain reaction (q-PCR).</p><p><b>RESULTS</b>VSMCs were promoted by Ang II at 10(-6) mol/L for 24 h. Compared with the normal group, the proliferative activity and migration quantity of VSMCs obviously increased, and DNA methylation level obviously decreased (P < 0.05, P < 0.01). Compared with the model group, the cell growth, proliferative activity and migration quantity of VSMCs obviously decreased and the general DNA methylation level increased in the folate group and the PQR group (P < 0.05, P < 0.01). Compared with the normal group, the mRNA expression of DNMT1 decreased in the model group (P < 0.01). Compared with the model group, mRNA expression of DNMT1 in Ang II induced VSMCs was obviously enhanced in the folate group and the PQR group (P < 0.01).</p><p><b>CONCLUSIONS</b>PQR could inhibit Ang II induced proliferation and migration of VSMCs, and cause high genomic DNA methylation level. Changes of DNA methylation might be associated with DNMT1 expression.</p>

Effect of Pinggan Qianyang recipe on the expression of Tpx II HSP27 and ANXA1 in the hypothalamus of spontaneously hypertensive rats with hyperactivity of liver-YANG syndrome / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effect Pinggan Qianyang recipe on expression of Tpx, HSP27 and ANXA1 in the hypothalamus of spontaneously hypertensive rats (SHRs) with the hyperactivity of liver-YANG syndrome.@*METHODS@#A total of 30 SHRs were subjected to administration of Aconiti Praeparatae Decoction to establish the model of SHR with liver-YANG hyperactivity first, then they were randomly divided into three groups: the control group, the model group and the treatment group (n=10 per group). A total of 10 SD rats were served as the normal group. The rats in control group and treatment group were given Enalapril plus Pinggan Qianyang recipe for four weeks. The change of behavior and blood pressure of rats were monitored. RT-PCR and Western-blot were performed to detect the expression of Tpx II, HSP27 and ANXA1 mRNA and protein in the hypothalamus, respectively.@*RESULTS@#Compared with the normal SD rats, the heart rate, blood pressure and grade of irritability were significantly increased while rotation endurance time was dramatically reduced in the SHR model with liver-YANG hyperactivity (P<0.01), these changes were reversed by the application of Enalapril plus Pinggan Qianyang recipe. Compared with the normal SD rats, the protein and mRNA expression of Tpx II and ANXA1 in the model group were significantly upregulated (P<0.01) while the HSP27 was significantly downregulated (P<0.01). Compared with the model group, the protein and mRNA expression of Tpx II and ANXA1 in the control group or treatment group were significantly decreased (P<0.05 or P<0.01) while HSP27 was significantly increased (P<0.05 or P<0.01). Compared with the control group, the expression of Tpx II and ANXA1 protein in treatment group were significantly reduced (P<0.05 or P<0.01).@*CONCLUSION@#Pinggan Qianyang recipe can improve the blood pressure and behavior in SHRs with hyperactivity of Liver-YANG syndrome, which might be related to the regulation of Tpx, HSP27 and ANXA1 expression in hypothalamuses.

The effect of hyaluronic acid on conception rate of ectopic patients who underwent laparoscopic surgery treatment / 中国基层医药

Objective To observe the clinical effect of hyaluronic acid on conception rate of ectopic pregnancy patients who received conservative tubal operation under laparoscopy.Methods 100 patients with ectopic pregnancy received conservative tubal operation under laparoscopy treatment were selected.Hyaluronic acid was used in 50 cases (the treatment group),unused anti blocking preparations for the control group in 50 cases.Then patients were followed up on the tubal patency,and observed pregnancy after 1 year.Results The rate of tubal patency after menstrual clean was 84％ in the treatment group,which was significantly higher than 60％ in the control group(x2 =7.155,P ＜ 0.05).In the treatment group,40 cases were intrauterine pregnancy,4 cases were ectopic pregnancy,6 cases were infertility.The rate of intrauterine pregnancy after 1 year was 80％ in the treatment group,which was significantly higher than 56％ in the control group(x2 =6.65,P ＜ 0.05).Conclusion Hyaluronic acid is effective in preventing tubal adhesion in the ectopic pregnancy patients who received laparoscopic conservative surgery,and can improve the postoperative tubal patency rate and conception rate.

Protective effect of ginsenoside Rg1 again PC-12 cells in OGD injury through mTOR/Akt/FoxO3 signaling pathway / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the protective effect of ginsenoside Rg1 on oxygen-glucose deprivation (OGD) in PC-12 cells, and preliminarily discuss the potential molecular mechanism of mTOR/Akt/FoxO3 signaling pathway.</p><p><b>METHOD</b>The OGD PC-12 cell model was established. The cell viability was measured by MTT assay. After the pretreatment with Rg1 with the concentration of 10, 20, 40 micromol x L(-1) for 24 h, the cell viability was observed. Lactate dehydrogenase (LDH) release, superoxide dismutase (SOD) ac- tivity and malondialdehyde (MDA) level were detected by colorimetry assay. mTOR, p-Akt(ser473), p-Akt(tjr308), Akt, p-FoxO3, FoxO3 in cytoplasm and nucleus, and total FoxO3 protein expression were detected by Western blot assay.</p><p><b>RESULT</b>OGD could significantly in- hibit cell proliferation in 4-24 h in a time-dependent manner. After pretreatment for 24 h, Rg1 (20, 40 micromol x L(-1)) could notably elevate the cell viability and SOD viability and reduce the LDH release and MDA content. Besides, Rg1 also inhibited OGD-induced mTOR and p-Akt(ser473) decreases. After treatment for 6 h, OGD could reduce FoxO3 phosphorylation and promote FoxO3 in cytoplasm. This data suggested that Rg1 could protect PC-12 cell injury through mTOR/p-Akt/FoxO3 signaling pathway.</p><p><b>CONCLUSION</b>Ginsenoside Rg1 could attenuate OGD-induced PC-12 cell injury. Its action mechanism may be closely related to activation of mTOR/p-Akt/FoxO3 signaling pathway.</p>

Differential expression of microRNA in endothelial cells incubated with serum of hypertension patients with blood-stasis syndrome / 中国结合医学杂志

<p><b>OBJECTIVE</b>To screen out blood-stasis syndrome (BSS)-associated microRNA and therefore determine the possible target for treating hypertension.</p><p><b>METHODS</b>A high-energy sequencing method and digital gene expression sequencing theory were adopted to sequence microRNA (miRNA) and messenger RNA (mRNA), and to determine differential expression in human umbilical vein endothelial cells incubated with serum samples from hypertension patients with or without BSS, and healthy controls. The results were confirmed using gene prediction software.</p><p><b>RESULTS</b>A total of 13 miRNAs and 11 mRNAs showed statistical difference both in the BSS/normal groups and BSS/non-BSS groups, respectively. Four pairs of target mRNA/miRNA were identified: FRMD4A/hsa-miR-34a, MAP3K14/hsa-miR-34a, PER1/hsa-miR-34a, and FGF2/hsa-miR-132.</p><p><b>CONCLUSION</b>Four mRNA/miRNA pairs mentioned above seem to be involved in pathogenesis and maintenance of hypertension with BSS.</p>

Constructing protein-protein interaction network of hypertension with blood stasis syndrome via digital gene expression sequencing and database mining / 中西医结合学报

<p><b>OBJECTIVE</b>To construct a protein-protein interaction (PPI) network in hypertension patients with blood-stasis syndrome (BSS) by using digital gene expression (DGE) sequencing and database mining techniques.</p><p><b>METHODS</b>DGE analysis based on the Solexa Genome Analyzer platform was performed on vascular endothelial cells incubated with serum of hypertension patients with BSS. The differentially expressed genes were filtered by comparing the expression levels between the different experimental groups. Then functional categories and enriched pathways of the unique genes for BSS were analyzed using Database for Annotation, Visualization and Integrated Discovery (DAVID) to select those in the enrichment pathways. Interologous Interaction Database (I2D) was used to construct PPI networks with the selected genes for hypertension patients with BSS. The potential candidate genes related to BSS were identified by comparing the number of relationships among genes. Confirmed by quantitative reverse transcription-polymerase chain reaction (qRT-PCR), gene ontology (GO) analysis was used to infer the functional annotations of the potential candidate genes for BSS.</p><p><b>RESULTS</b>With gene enrichment analysis using DAVID, a list of 58 genes was chosen from the unique genes. The selected 58 genes were analyzed using I2D, and a PPI network was constructed. Based on the network analysis results, candidate genes for BSS were identified: DDIT3, JUN, HSPA8, NFIL3, HSPA5, HIST2H2BE, H3F3B, CEBPB, SAT1 and GADD45A. Verified through qRT-PCR and analyzed by GO, the functional annotations of the potential candidate genes were explored.</p><p><b>CONCLUSION</b>Compared with previous methodologies reported in the literature, the present DGE analysis and data mining method have shown a great improvement in analyzing BSS.</p>

Influence of breast milk on secretion of Ghrelin and Peptide YY in rats / 中华实用儿科临床杂志

Objective To study the effect of breast milk on secretion of Ghrelin and Peptide YY (PYY)during different periods in SD rats.Methods Sprague-Dawley (SD) rats were randomly divided into breast milk-fed group(BG) and formula-artificial fed group(FG) with 20 in each group.After 21 days both of the 2 groups were fed by same forage.Ten of each group were experimentized in day 21 equivalently weaning period,and rest rats were experimentized at the 50th day equivalently childhood.Real-time PCR was used to determine the mRNA of PYY,Ghrelin from gastric,colon.Immunohistochemistry was used to determine Ghrelin,PYY protein expression in the gastrointestinal tract tissues.Results There were no difference between body weight and gastrointestinal mucosal development of 2 rat groups in day 21 and day 50(P ＞0.05),mRNA and protein expression of Ghrelin and PYY in breast milk-fed group were higher than formula-artificial fed group in both day 21 and day 50 (all P ＜ 0.05).Conclusions The Ghrelin and PYY levels of breast milk-fed rats is higher than formula-artificial fed ones,and this phenomenon continues to their childhood.Breast milk protects offspring from obesity by influencing the secretion of brain-gut peptide and has long-term consequences on the regulation of food intake and energy balance from neonatal period to their later life.

Test-retest reliability of Nordic Musculoskeletal Questionnaire in nurses / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To evaluate the test-retest reliability of Nordic Musculoskeletal Questionnaire in nurses and to provide supplementary data for evaluating the reliability of the questionnaire in different occupational populations.</p><p><b>METHODS</b>Nordic Musculoskeletal Questionnaire was translated into Chinese according to the Guidelines for the process of cross-cultural adaptation of self-report measures. We carried out a study to examine the reliability of Chinese-version Nordic Questionnaire among Chinese nurses. This study was conducted in 120 nurses recruited from our hospital, who underwent questionnaire survey twice within one week. The test-retest reliability of questionnaire was analyzed.</p><p><b>RESULTS</b>The Chinese-version Nordic Questionnaire showed a high test-retest reliability, with a Kappa coefficient of 0.72∼1.00.</p><p><b>CONCLUSION</b>The Chinese-version Nordic Questionnaire has a high test-retest reliability in nurses, so it can be used for the screening and epidemiological investigation of musculoskeletal disorders in this population.</p>

Effect of Wnt signaling suppression on gefitinib in non small cell lung cancer cell lines / 中华病理学杂志

<p><b>OBJECTIVE</b>To explore the effect of Wnt signaling suppression on proliferation of non small cell lung cancer to gefitinib, and its related mechanisms.</p><p><b>METHODS</b>PC9 and PC9/AB2 cells of both gefitinib sensitive and resistant were treated with different concentrations of gefitinib, and the proliferation index was measured using CCK8 kit. The members of Wnt signaling pathway were detected by Western blot. Dual luciferase reportor gene assay (TOP Flash) was used to document the transcriptional level of β-catenin. β-catenin siRNA was transfected into PC9/AB2 cells to suppress the Wnt signaling transcription, followed by treatment with different concentrations of gefitinib. Western blot was then used to detect the expression of EGFR and its downstream signaling after inhibit the expression of β-catenin.</p><p><b>RESULTS</b>Treating with different concentrations of gefitinib, the resistance of PC9/AB2 cells to gefitinib was significantly increased (P < 0.05). The members of Wnt signaling expressed at higher level in PC9/AB2 cells than in PC9 cells (t = 24.590, P = 0.000). TOP Flash examination showed that the endogenous transcriptional activity of Wnt signaling was higher in PC9/AB2 cell than that in PC9 cell (t = 4.983, P = 0.008). Compared with the negative control group, apoptotic rate and sensitivity to gefitinib significantly increased in interfered group (P < 0.05). The expression of p-ERK1/2 significantly decreased after Wnt signaling suppression, although other proteins showed no significant alterations.</p><p><b>CONCLUSION</b>Suppressing the activity of Wnt signaling can partly reverse the celluar resistance to gefitinib in non small cell lung cancer.</p>

Association of nicotinamide adenine dinucleotide phosphate oxidase p22phox gene 549C>T polymorphism with coronary artery disease / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The p22phox is a critical component of the superoxide-generating vascular nicotinamide adenine dinucleotide phosphate (NADPH) oxidase. Several polymorphisms in p22phox gene are studied for their association with cardiovascular diseases. However, no publication is available to assess the relation of 549C > T polymorphism in p22phox gene to coronary artery disease (CAD) risk. This study was to investigate the effect of the p22phox gene 549C > T polymorphism on CAD risk.</p><p><b>METHODS</b>Hospital-based case-control study was conducted with 297 CAD patients and 343 healthy persons as the control group. Polymerase chain reaction and pyrosequencing using PSQ 96 MA Pyrosequencer (Biotage AB) were used to detect the polymorphisms. Multiple Logistic regression model was used to adjust the potential confounders and to estimate odds ratio (OR) with 95% confidence intervals (CIs).</p><p><b>RESULTS</b>The observed genotype frequencies of this polymorphism obeyed the Hardy-Weinberg equilibrium in both cases (P = 0.439) and controls (P = 0.668). The frequency of mutant genotypes (TT + CT) in cases (41.08%) was higher than that in controls (36.73%) with an OR = 1.20 (95%CI = 0.87-1.65). After the adjustment of the potential confounders, there was a significant association of the mutant genotypes with increased risk of CAD (OR = 1.57, 95%CI = 1.01-2.46, P = 0.047).</p><p><b>CONCLUSIONS</b>The mutant genotypes of the p22phox gene 549C > T polymorphism had a significant effect on the increased risk of CAD in this studied population.</p>

Enhance efficiency of adenovirus on AAV transfering human RPE cells / 中华实验眼科杂志

Background Adeno-associated virus-based vector is one of most efficient vehicles.It presents with a long-term and efficient transfer and expression of therapeutic genes with minimal toxicity.But its delayed-and low-efficient transgene expression limits the application of AAV vector.To explore an improving method of AAV infecting RPE cells is the hot spot. Objeetive Present study was to investigate whether adeno-associated virus (AAV)combined with low dose non-replieable adenovirus(Ad-null)can enhance its infection efficieney on RPE ceils in vitro. Methods Human RPE cells were isolated from the donate eyeballs under the approval of the Ethic Committee of this hospital.The cells were cultured in DMEM containing 10%fetal bovine serum.AAV particles with enhanced green fluorescence protein(EGFP)were added into the medium alone or in combination with different amount of adenovirus for 30 days.The cells were detected under the fluorescence microscope,and the protein expression levels of report gene EGFP in RPE cells were analyzed with Western blotting assay. Results Melanin granules could be found in cultured RPE cells.EGFP was expressed in RPE cells at 2 days after AAV-EGFP infection and peaked at 12 days and remained for about 3-week duration,showing the green influorescence under the influorescence mwroscope.After the cells were infected by AAV2-EGFP with 0.01 to 1000 MOI Ad-null respectively,the number of cells with green influorescence was obviously increased with the enhanced infiuorescence intensity.The enhance of the infection efficiency began in the 0.1 MOI Ad-null group and peaked in 10 MOI Ad-null group.Dead cells were exhibited in the 100 or more MOI Ad-nulor group.Western blotting assay demonstrated that the protein expression level of EGFP in RPE cells enhanced significantly in 1 and 10 MOI Ad-null groups compared with only AAV infection group. Conclusion These finding suggested that the infection efficiency of AAV can be improved significantly when it is used with low dose Ad-null in vitro.This offers a basis for further study of gene therapy.

Low dosage 5-fluorouracil increases the transfection efficiency of Ad/VEGF-A in mouse lung carcinoma cell line LA795 and inhibits tumor growth / 癌症

<p><b>BACKGROUND AND OBJECTIVE</b>Adenovirus vectors were widely used in gene therapy for tumors. We used adenovirus vector to transfer small interfering RNA (siRNA) against vascular epithelium growth factor A (VEGF-A) molecules to mouse lung adenoma LA795 cells and used low dose of chemotherapeutic drugs to further elevate the infection efficiency of adenovirus vector in and therapeutic effect of RNAi on tumor cells.</p><p><b>METHODS</b>LA795 cells were infected by Ad/EGFP and treated with different dosages of gemcitabin, epirubicin, cisplatin, or 5-fluorouracil (5-FU). Cells were observed under fluorescence microscope continuously using green fluorescent protein (GFP) as the reporter gene. The percentage of GFP-positive cells and fluorescent intensity were tested by flow cytometry to determine optimum concentrations of drugs. Ad/siVEGF-A containing VEGF-A siRNA was constructed. Real-time PCR and ELISA were applied to measure the expression level of VEGF-A after LA795 cells were infected by Ad/siVEGF-A and treated with 5-FU. The combination of Ad/siVEGF-A and 5-FU was also applied in treating subcutaneous tumor in mice.</p><p><b>RESULTS</b>Low dose of 5-FU elevated the Ad/EGFP infection in LA795 cells significantly, and also enhanced the effect of Ad/siVEGF-A in down-regulating VEGF-A mRNA and protein levels in tumor cells. When used in tumor in vivo, the combination strategy repressed tumor growth effectively.</p><p><b>CONCLUSION</b>Low dose of 5-FU can enhance the capability of adenovirus infecting tumor cells and promote the efficiency of gene therapy by adenovirus.</p>