Caspase-1/-11 participates in LPS-induced sepsis-associated acute kidney injury by cleaving GSDMD / 生理学报

The present study was aimed to investigate whether Gasdermin D (GSDMD)-mediated pyroptosis participated in lipopolysaccharide (LPS)-induced sepsis-associated acute kidney injury (AKI), and to explore the role of caspase-1 and caspase-11 pyroptosis pathways in this process. The mice were divided into four groups: wild type (WT), WT-LPS, GSDMD knockout (KO) and KO-LPS. The sepsis-associated AKI was induced by intraperitoneal injection of LPS (40 mg/kg). Blood samples were taken to determine the concentration of creatinine and urea nitrogen. The pathological changes of renal tissue were observed via HE staining. Western blot was used to investigate the expression of pyroptosis-associated proteins. The results showed that the concentrations of serum creatinine and urea nitrogen in the WT-LPS group were significantly increased, compared with those in the WT group (P < 0.01); whereas serum creatinine and urea nitrogen in the KO-LPS group were significantly decreased, compared with those in the WT-LPS group (P < 0.01). HE staining results showed that LPS-induced renal tubular dilatation was mitigated in GSDMD KO mice. Western blot results showed that LPS up-regulated the protein expression levels of interleukin-1β (IL-1β), GSDMD and GSDMD-N in WT mice. GSDMD KO significantly down-regulated the protein levels of IL-1β, caspase-11, pro-caspase-1, caspase-1(p22) induced by LPS. These results suggest that GSDMD-mediated pyroptosis is involved in LPS-induced sepsis-associated AKI. Caspase-1 and caspase-11 may be involved in GSDMD cleavage.

Analysis of common pathogenic gene mutations of arrhythmogenic right ventricular cardiomyopathy in Yunnan unexplained sudden death cases / 中华地方病学杂志

Objective:To analyze common pathogenic gene mutations of arrhythmogenic right ventricular cardiomyopathy (ARVC) in Yunnan unexplained sudden death (hereinafter referred to as Yunnan sudden death) cases, and explore the etiological relationship between Yunnan sudden death and ARVC.Methods:Four typical Yunnan sudden death affected counties (cities) were selected as investigation sites. Cryopreserved autopsy cardiac cavity blood samples were collected from Yunnan sudden death cases ( n = 3), and peripheral venous blood samples were harvested from their relatives (first, second, third and immediate degree of kinship, n = 67) and control population ( n = 49). The DNA of blood samples was extracted for amplification and sequencing of 97 exons of 5 common ARVC desmosomal protein [desmoplakin (DSP), desmocollin-2 (DSC2), desmoglein-2 (DSG2), plakophilin-2 (PKP2) and junction plakoglobin (JUP)] genes, and genetic lineage of Yunnan sudden death cases was investigated. Results:A total of 17 gene mutation sites were discovered in Yunnan sudden death cases and their relatives, with 6, 5, 4, 1 and 1 in the DSP, DSC2, DSG2, PKP2 and JUP genes, which were not found in the control population. Among them, 9 were newly discovered mutation sites and 8 were reported mutation sites. The DSP gene exon 24 c.8472 G>C, a pure contractual sense mutation, was common in the relatives of 4 cases in the same family surveyed; and one immediate relative carried a deletion mutation at c.2368 - 2370 of exon 15 of DSC2 gene.Conclusion:Yunnan sudden death cases and their relatives carry mutations in the ARVC desmosomal protein DSP, DSC2, DSG2, PKP2, and JUP genes, and the onset of some Yunnan sudden death may be associated with mutations in the ARVC desmosomal protein genes.

Ultrasound-guided stellate ganglion block accelerates postoperative gastrointestinal function recovery following laparoscopic radical gastrectomy for gastric cancer / 南方医科大学学报

OBJECTIVE@#To explore the effects of ultrasound-guided stellate ganglion block (SGB) on perioperative stress response, gastrointestinal hormones and postoperative gastrointestinal function recovery in patients undergoing laparoscopic radical gastrectomy for gastric cancer.@*METHODS@#This study was conducted among 60 American Society of Anesthesiologists (ASA) class II-III patients with gastric cancer (regardless of gender, aged 35-75 years with BMI of 18.5-26 kg/m2) undergoing elective laparoscopic radical gastrectomy. The patients were randomized into experimental group (S group, n=30) and control group (NS group, n=30). In S group, SGB at the C6 level of the right cervical spine was performed under ultrasound guidance 15 min before induction of anesthesia by injection of 7 mL 0.5% ropivacaine; the patients in NS group received injections of normal saline in the same manner. Peripheral venous blood samples were collected before SGB (T1), after surgery (T2), and on the 2nd and 6th days after surgery (T3 and T4) for determination of the levels of motitin (MOT), vasoactive intestinal peptide (VIP), cortisol (COR), and blood glucose (GLU). Intraoperative usage of sufentanil, recovery rate of intestinal sounds at 36, 48, 60, 72, 84 and 96 h after operation and the time of first passage of flatus were recorded and compared between the two groups.@*RESULTS@#There was no significant difference in the total amount of sufentanil consumption between the two groups. Compared with those in NS group, the patients in S group had significant lower COR and VIP levels (P < 0.05) and higher MOT level (P < 0.05) at T2, T3 and T4. Glu level at T2 and T3 was also significantly lower in S group (P < 0.05). The recovery rates of intestinal sounds at 36, 48, 60, 72 and 84 h after surgery were significantly higher (P < 0.05) and the time of the first passage of flatus was earlier in S group than in NS group (P < 0.05).@*CONCLUSION@#In patients with gastric cancer undergoing laparoscopic radical gastrectomy, ultrasound-guided SGB can reduce postoperative stress level, promote the recovery of gastrointestinal hormone secretion, and accelerate postoperative recovery of gastrointestinal functions.

Computer simulation of molecular docking between methylene blue and some proteins of Porphyromonas gingivalis / 北京大学学报(医学版)

OBJECTIVE@#To study the binding target of photosensitizer and bacteria in antimicrobial photodynamic therapy with computer-simulated target prediction and molecular docking research methods and to calculate the binding energy.@*METHODS@#The protein names of Porphyromonas gingivalis (Pg) were obtained and summarized in Uniprot database and RCSB PDB database; the structure diagrams of methy-lene blue were screened in SciFinder database, PubChem database, ChemSpider database, and Chemical Book, and ChemBioDraw software was used to draw and confirm the three-dimensional structure for target prediction and Cytoscape software was used to build a visual network diagram; a protein interaction network was searched and built between the methylene blue target and the common target of Pg in the String database; then we selected FimA, Mfa4, RgpB, and Kgp K1 proteins, used AutoDock software to calculate the docking energy of methylene blue and the above-mentioned proteins and performed molecular docking.@*RESULTS@#The target prediction results showed that there were 19 common targets between the 268 potential targets of methylene blue and 1 865 Pg proteins. The 19 targets were: groS, radA, rplA, dps, fabH, pyrG, thyA, panC, RHO, frdA, ileS, bioA, def, ddl, TPR, murA, lepB, cobT, and gyrB. The results of the molecular docking showed that methylene blue could bind to 9 sites of FimA protein, with a binding energy of -6.26 kcal/mol; with 4 sites of Mfa4 protein and hydrogen bond formation site GLU47, and the binding energy of -5.91 kcal/mol, the binding energy of LYS80, the hydrogen bond forming site of RgpB protein, was -5.14 kcal/mol, and the binding energy of 6 sites of Kgp K1 protein and the hydrogen bond forming site GLY1114 of -5.07 kcal/mol.@*CONCLUSION@#Computer simulation of target prediction and molecular docking technology can initially reveal the binding, degree of binding and binding sites of methylene blue and Pg proteins. This method provides a reference for future research on the screening of binding sites of photosensitizers to cells and bacteria.

Correlation between preoperative serum γ-gultamyl transferase level and postoperative survival in patients with renal cell carcinoma / 肿瘤研究与临床

Objective:To investigate the correlation between preoperative serum γ-glutamyl transferase (GGT) level and the postoperative survival of patients with renal cell carcinoma.Methods:The clinical data of 235 patients with renal cell carcinoma who underwent nephrectomy from December 2005 to December 2011 in the Affiliated Hospital of China University of Mining and Technology were retrospectively analyzed. According to the preoperative serum GGT level at 1.5 times the upper limit of normal value 90 U/L (the upper limit of normal value of samples in this study was 60 U/L), patients were divided into 2 groups: GGT ≤ 90 U/L group (218 cases) and GGT>90 U/L group (17 cases). The correlation between GGT and clinicopathological characteristics as well as postoperative survival of patient with renal cell carcinoma was also analyzed.Results:There were statistically significant differences in gender, age, the level of the aspartate aminotransferase (AST), the level of alanine aminotransferase (ALT), the neutrpphil-to-lymphocyte ratio (NLR), lymph node metastasis, distant metastasis, neoplasm staging between GGT ≤ 90 U/L group and >90 U/L group (all P < 0.05). Kaplan-Meier survival analysis showed that the median overall survival time was 84 months and 54 months, respectively in GGT ≤ 90 U/L group and GGT > 90 U/L group, and the difference was statistically significant of both groups ( χ2 = 4.334, P = 0.037). Univariate Cox proportional risk regression model showed that pathologic type, pathology T staging, preoperative GGT level, lactate dehydrogenase (LDH) level, ALT level were influencing factors of postoperative overall survival in patient with renal cell carcinoma (all P < 0.05). The multivariate Cox regression model analysis showed that the pathological type ( HR = 2.323, 95% CI 1.228-4.396, P = 0.010), GGT level ( HR = 2.406, 95% CI 1.077-5.376, P = 0.032), LDH level ( HR = 2.320, 95% CI 1.080-4.981, P = 0.031) were independent influencing factors of postoperative overall survival in patient with renal cell carcinoma. Conclusions:Preoperative elevated serum GGT level is associated with poor prognosis of patients with renal cell carcinoma, and the monitoring of it may help to evaluate the prognosis of patients and provide guidelines for individual treatment method.

Comparative efficacy and safety of first-line EGFR-TKIs for advanced non-small cell lung cancer：a network meta-analysis / 中华疾病控制杂志

Objective To compare the efficacy and safety of gefitinib, erlotinib, and afatinib in the first-line treatment of advanced non-small cell lung cancer (NSCLC). Methods PubMed, EMBASE, and The Cochrane Library were searched to identify the relevant literatures published from December 2008 to December 2018. Bayesian network meta-analysis was carried out to rank the three treatments. Results A total of ten eligible studies involving 2275 patients were enrolled. In terms of efficacy, the surface under the cumulative ranking (SUCRA) indicated that erlotinib performed best in progression-free survival(PFS)(0.88), afatinib performed best in objective response rate(ORR)(0.82) and disease control rate(DCR) (0.86), gefitinib performed worst in PFS (0.45), ORR(0.42), and DCR(0.45). For safety, the differences of grade 3 or 4 adverse events rate (OR=0.29,95%CI:0.08-0.98) and discontinuation rate(OR=0.14,95%CI:0.01-0.8) between erlotinib and the platinum-based doublet chemotherapy were statistically significant. The ranking results also supported that erlotinib was the safest. SUCRA results suggested that gefitinib (0.31) had a lower grade 3 or 4 adverse events rate than afatinib (0.57), and the possibility of discontinuation in gefitinib (0.44) was similar to that of afatinib (0.41). Conclusion Erlotinib might be the preferred first-line treatment for advanced NSCLC after weighing and balancing the benefits and risks.

HPLC Fingerprint of Bletillae Formosanae Rhizoma / 中国实验方剂学杂志

Objective: To study the chemical constituents of Bletillae Formosanae Rhizoma and the distribution characteristics of active components in the fingerprint by establishing its high performance liquid chromatography (HPLC) fingerprints. Method: HPLC was used to establish the fingerprint of Bletillae Formosanae Rhizoma. Four reference substances,i.e. militarine,coelonin,4-methoxy-9,10-dihydrophenanthrene-1,2,7-triol and batatasin Ⅲ were used to identify chromatographic peaks. The fingerprints of 17 batches of Bletillae Formosanae Rhizoma fingerprints were analyzed and compared by "Computer-aided-similarity evaluation soft" and stoichiometry,and then compared with the fingerprint of Bletillae Rhizoma. Result: The established HPLC fingerprint method of Bletillae Formosanae Rhizoma showed good repeatability and stability. 20 common peaks were marked,four of which were identified by reference substances; militarine was No.8 common peak,and others corresponded to No. 10, No. 14 and No. 18 common peaks. Results showed that the similarities of samples except S4 were higher than 0.85, but the relative peak area of common peaks was quite different. Within the cluster distance 10,the samples are clustered into 5 categories, reflecting certain origin correlation. Principal component analysis (PCA) showed that the difference in samples was mainly caused by the common peaks located after No. 9 peak,where chemical constituents such as bibenzyl and dihydrophenanthrene were distributed. Bletillae Formosanae Rhizoma and Bletillae Rhizoma showed similar chemical constituents. Conclusion: The method provided a theoretical basis for the further clinical application and quality control of Bletillae Formosanae Rhizoma,as a substitute for Bletillae Rhizoma.

Diffusion kurtosis imaging characteristics of brain gray matter nucleus microstructure in Parkinson's disease with different movement disorder subtypes / 中华行为医学与脑科学杂志

Objective@#To investigate the difference in the microstructure of gray matter nucleus in different movement subtypes of Parkinson’s disease (PD) by diffusion kurtosis imaging (DKI) technique, and to analyze the correlation with clinical manifestations.@*Methods@#Ninety-seven patients with PD and 83 healthy controls performed conventional MRI sequence and DKI sequence scan. The PD patients were classified into gait disorder subtype (PIGD, n=57) and tremor dominant subtype (TD, n=40)subtypes according to motor symptoms. Fractional anisotropy (FA), mean diffusivity (MD), axial diffusivity (Da), radial diffusivity(Dr), mean kurtosis (MK), axial kurtosis (Ka) and radial kurtosis (Dr) maps and data were generated by software after processing. DKI was performed for all subjects and data was collected from different brain regions in both hemispheres, including red nucleus(RN), substantia nigra pars reticulate(SNr), substantia nigra pars compacta(SNc), putamen(PUT), globus pallidus(GP), head of caudate nucleus (CN)and thalamus(THA).@*Results@#TD showed a higher MMSE score(P=0.019), but lower modified Hoehn-Yahr score than that in PIGD (P<0.001), there was no significant difference of age of onset, sex, limbs of onset or disease duration between two PD subgroups. Compared with healthy controls, both TD and PIGD showed down-regulated MD, Da and Dr and up-regulated Ka values(P<0.001); MK(0.83±0.26, 0.80±0.18) was increased in SNr both in TD and PIGD, while SNc, PUT and GP (0.84±0.20, 0.75±0.07, 0.81±0.14)were decreased only in TD (P=0.017, P=0.010, P=0.020, P<0.001, P=0.002). The Kr values of PUT and CN(0.71±0.17, 0.72±0.14) were reduced in PIGD, while CN(0.70±0.14) were reduced in TD respectively (P=0.002, P=0.031, P=0.007). The MK was lower in TD than that in PIGD (t=-2.214, P=0.029), and no significant difference was found in other grey matter nuclei between TD and PIGD(P>0.05). Moreover, there was no significant correlation between DKI value and disease duration, MMSE score or Hoehn-Yahr scale (P>0.05) in TD and PIGD.@*Conclusion@#There is heterogeneity of clinical symptoms between these two subgroups of PD. DKI can quantify the microstructural changes of grey matter nucleus in different type PD patient.

Diffusion kurtosis imaging characteristics of brain gray matter nucleus microstructure in Parkinson's disease with different movement disorder subtypes / 中华行为医学与脑科学杂志

Objective To investigate the difference in the microstructure of gray matter nucleus in different movement subtypes of Parkinson’s disease (PD) by diffusion kurtosis imaging ( DKI) technique, and to analyze the correlation with clinical manifestations. Methods Ninety-seven patients with PD and 83 healthy controls performed conventional MRI sequence and DKI sequence scan. The PD patients were classi-fied into gait disorder subtype (PIGD,n=57) and tremor dominant subtype (TD,n=40)subtypes according to motor symptoms. Fractional anisotropy (FA),mean diffusivity (MD),axial diffusivity (Da),radial diffu-sivity(Dr),mean kurtosis (MK),axial kurtosis (Ka) and radial kurtosis (Dr) maps and data were genera-ted by software after processing. DKI was performed for all subjects and data was collected from different brain regions in both hemispheres,including red nucleus(RN),substantia nigra pars reticulate( SNr),sub-stantia nigra pars compacta(SNc),putamen(PUT),globus pallidus(GP),head of caudate nucleus (CN)and thalamus(THA). Results TD showed a higher MMSE score(P=0. 019),but lower modified Hoehn-Yahr score than that in PIGD (P<0. 001),there was no significant difference of age of onset,sex,limbs of onset or disease duration between two PD subgroups. Compared with healthy controls, both TD and PIGD showed down-regulated MD,Da and Dr and up-regulated Ka values(P<0. 001); MK(0. 83±0. 26,0. 80±0. 18) was increased in SNr both in TD and PIGD,while SNc,PUT and GP (0. 84± 0. 20,0. 75± 0. 07,0. 81± 0. 14) were decreased only in TD (P=0. 017,P=0. 010,P=0. 020,P<0. 001,P=0. 002). The Kr values of PUT and CN(0. 71±0. 17,0. 72±0. 14) were reduced in PIGD,while CN(0. 70±0. 14) were reduced in TD re-spectively (P=0. 002,P=0. 031,P=0. 007). The MK was lower in TD than that in PIGD (t=-2. 214,P=0. 029),and no significant difference was found in other grey matter nuclei between TD and PIGD ( P>0. 05). Moreover,there was no significant correlation between DKI value and disease duration,MMSE score or Hoehn-Yahr scale (P>0. 05) in TD and PIGD. Conclusion There is heterogeneity of clinical symptoms between these two subgroups of PD. DKI can quantify the microstructural changes of grey matter nucleus in different type PD patient.

Evaluation of Literature Quality of Randomized Controlled Trials on Wrist-ankle Acupuncture for Low-back Pain / 中国中医药信息杂志

Objective To evaluate the literature quality of randomized controlled trials (RCTs) on wrist-ankle acupuncture (WAA) for low-back pain in recent ten years; To analyze the existing problems in the clinical research;To provide corresponding improvement suggestions. Methods A computer-based retrieval was performed to search out the reports of RCTs on WAA for low-back pain from CNKI, Wanfang Data, CBM, Chongqing VIP, PubMed, and Cochrane Library was retrieved by computers. The search scope was January 1, 2007 - December 31, 2016. The 25 items in the Consolidated Standards of Reporting Trials (CONSORT) Statement and 6 items of the Standards for Reporting Interventions in Clinical Trials of Acupuncture (STRICTA) were consulted for assessing the literature quality of RCTs on WAA for low-back pain. Results 18 articles were included. There were many problems about literature of RCTs on WAA for low-back pain, mainly including that the type of test design was not clear, outcome indicators were incomplete, random method reports were not specific, accepted diagnostic criteria and efficacy criteria were not used, reporting interventions were incomplete. Conclusion Recently, literature quality of RCTs on WAA for low-back pain is low. It is suggested that CONSORT Statement and STRICTA should be taken into consideration in the conducting and reporting of RCTs on WAA for low-back pain, and the report quality of clinical research in this field should be improved.

Analysis of One Case of Senior Patient with Deep Vein Thrombosis Occurred Retroperitoneal Bleeding during Anticoagulant Therapy / 中国药师

Objective:To improve the recognition of bleeding risk in the anticoagulant process by analyzing one case of senior patient with deep vein thrombosis occurred severe bleeding during the anticoagulant therapy.Methods:The possible causes of hemorrhage were analyzed after the brief introduction of medical history,lab test report and treatment process of the patient.Results:The reasons of hemorrhage might be as follows:① the combination of warfarin and dalteparin induced the adverse drug reaction;② albumin decreased during the treatment process resulting in the effect enhancement of warfarin;③ the clearance of warfarin decreased resulting from the low creatinine clearance of the elderly patient.Conclusion:Clinical anticoagulant practice should be more careful in senior patients.The frequency of related laboratory tests should be increased in order to find potential risks timely.

Comparison of Three Methods for Culturing Rheumatoid Arthritis Synovial Fibroblasts / 昆明医科大学学报

Objective To compare the three kinds of methods for in vitro primary culturing of rheumatoid arthritis synovial fibroblast-like cells (RASFs), in order to get fast and effective culture methods. Methods Synovial tissue from RA synovial arthroscopic resection were treated by collagenase digestion method, modified tissue culture method, double enzyme digestion method respectively. By using an inverted phase contrast microscope, cell morphology and growth characteristics were observed and identified with vimentin staining. Trypan blue was used to count the number of living cells after culturing for 14d. Results The three primary methods could successfully isolate and culture RASFs, and RASFs met the morphological characteristics of vimentin-positive cells>95%, namely, the proportion of RASFs cell confluence was 70% after 16-20days by the collagenase digestion method,whose cell confluence proportion reached 95%after 4 weeks;and the cell confluence proportion was above 70%after 10-14days by modified tissue culture method,and the cell confluence proportion reached 85%after 4 weeks by the double enzyme digestion method. The comparison of the viable cells number cultured same number of synovial tissue by the three methods show the viable cells number cultured by the modified tissue culture method were (1.60±0.08) ×106, those by the collagenase digestion method were (1.41±0.08) ×106, those by the double enzyme digestion method were (1.19 ±0.05) ×106, which were with significant difference among them (P<0.05) .The comparison of incubation time of RASFs primary cells showed it took (267.50±16.58) mins by the collagenase digestion method, (183.75 ±11.08) mins by the double enzyme digestion method, and 149.10 ±13.71mins by the modified tissue culture method, with significant differences (P<0.05) .Conclusion Modified tissue culture for RASFs is an efficient and fast culture method, the number and purity of RASFs can meet the requirements for biology experiments.

Establishment and Identification of Mouse Model of Philadelphia Chromosome Positive Acute Lymphoblastic Leukemia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To establish a mouse model of Phacute lymphoblastic leukemia (ALL) for providing a valuable tool to facilitate the researches on PhALL.</p><p><b>METHODS</b>CML-like mice were generated by transfection to bone marrow cells of BABL/c with Mig190 retrovirus. The PhALL mouse model was established by infusion of sorted CML like mouse-derived BCR-ABLB cells into the mice of same linege. Immonophenotypes, BCR-ABL transcription and expression of these leukemic cells were detected by flow cytometry, RT-PCR and Western blot respectively.</p><p><b>RESULTS</b>CML-like presentation appeared in the mice after Mig190 virus transfection. After infusion with sorted BCR-ABLB cells, all the receipted mice eventually presented the clinical signs of acute leukemia. Flow cytometry analysis showed that these leukemic cells were positive for CD19; both RT-PCR and Western blot indicated that this mouse model is consistent with PhALL phenotypecally and molecalarlly.</p><p><b>CONCLUSION</b>A novel method to establish PhALL mouse model has been developed successfully, and this model can provide useful tool for the study of PhALL.</p>

Effect of BRD4 Inhibitor GSK525762A on Proliferation and Apoptosis of SUP-B15 Cells In Vitro and Its Possible Mechanism / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the effect of BRD4 inhibitor GSK525762A on the proliferation and apoptosis of Philadelphia chromosome-positive acute lymphoblastic leukemia SUP-B15 cells and its mechanism.</p><p><b>METHODS</b>SUP-B15 cells were treated with different concentration of GSK525762A, the proliferation-inhibition curve was assayed and plotted by CCK-8 method, the cell viability and apoptosis were detected by flow cytometry with Annexin V and 7-AAD staining. The transcripts of anti-apoptotic genes C-MYC, CDK6 and BCL-2 were detected by real-time PCR.</p><p><b>RESULTS</b>GSK525762A could inhibit significantly SUP-B15 cell proliteration in dose-and time-dependent manner; GSK525762A treatment could induce apoptosis of SUP-B15 cells. The levels of C-MYC,CDK6 and BCL-2 mRNA transcripts in SUP-B15 cells were reduced in GSK525762A-treated group.</p><p><b>CONCLUSION</b>The GSK525762A can remarkably inhibit proliferation and induce apoptosis of SUP-B15 cells. The down-regulation of apoptosis-related genes C-MYC,CDK6 and BCL-2 may be involved in the process of apoptosis.</p>

Effect of ADAM10 Inhibitor GI254023X on Proliferation and Apoptosis of Acute T-Lymphoblastic Leukemia Jurkat Cells In Vitro and Its Possible Mechanisms / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the effect of ADAM10 inhibitor GI254023X on the proliferation and apoptosis of acute T-lymphoblastic leukemia Jurkat cells and its mechanisms.</p><p><b>METHODS</b>Jurkat cells were treated with different concentrations of GI254023X, the proliferation-inhibition curve was assayed and plotted by CCK-8 method, the cell viability and apoptosis was detected by flow cytometry with Annexin V and 7-AAD staining, the cleavage of Notch1 protein was determined by Western blot, the transcripts of anti-apoptotic genes BCL-2, MCL-1, BCL-xl and Notch1 target gene Hes-1 were detected by real-time PCR.</p><p><b>RESULTS</b>The GI254023X obviously inhibited the proliferation of Jurkat cells in concentration-dependent manner. As compared with the control group, the apoptosis of cells increased along with increment of GI254023X concentration. Compared with control group, the expression of Cleaved Notch1 was down-regulated while the expression of Notch1 was up-regulated in a time-dependent manner after the treatment with GI254023X. The levels of MCL-1 and Hes-1 mRNA transcripts in Jurkat cells were reduced in GI254023X treated group, but did not show obvious effect on the level of BCL-2 and BCL-xl mRNA transcripts.</p><p><b>CONCLUSION</b>GI254023X can remarkably inhibit proliferation and induce apoptosis of Jurkat cells. The inhibition of Notch1 activation and the down-regulation of apoptosis-related gene MCL-1 may be involved in the process of apoptosis.</p>

siRNA inhibition of survivin gene expression in rheumatoid arthritis synovial fibroblast proliferation and apoptosis / 中国生化药物杂志

Objective To study the targeting survivin small interfering RNA ( siRNA ) to inhibit proliferation and apoptosis survivin gene expression in rheumatoid arthritis synovial fibroblasts ( RASFs) .Methods RA patients were isolated and cultured in vitro synovial fibroblasts ( RASFs) , designed and synthesized siRNA targeting survivin and negative control, by liposome transfection RASFs cell; real-time quantitative polymerase chain reaction (PCR) and Western blot RASFs detect mRNA expression and protein levels of survivin.Tetrazolium blue (MTT) assay of cell proliferation;TUNEL assay apoptosis.Results The experimental group compared with the negative control siRNA group and control group, 48h after transfection of synovial fibroblasts survivin mRNA and protein expression levels were significantly decreased ( P<0.05 ) .The experimental group compared with the negative control siRNA group and control group, synovial fibroblast proliferation after transfection significantly decreased ( P<0.05 ) . After the experimental group transfected 24h, 48h, 72h growth inhibition rates were (11.5 ±2.6)%, (26.2 ±3.4)%, (47.6 ±4.1)%, at 72 hours after transfection most significant.The rate of apoptosis in experimental group (23.87 ±1.6)%, significantly higher than the negative control group (9.72 ± 1.15)% and the control group (8.70 ±1.09)% (all P<0.05).Conclusion siRNA targeting survivin expression levels through reducing survivin, inhibit synovial fibroblast proliferation and promotes apoptosis.

Effects of shRNA-mediated survivin silencing on proliferation and apoptosis of human umbilical vein endothelial cells / 重庆医学

Objective To investigate the effect of short hairpin RNA(shRNA) eukaryotic expression vector‐mediated silen‐cing of the survivin‐gene on proliferation and apoptosis of human umbilical vein endothelial cells(HUVEC) .Methods The shRNA vector targeting the survivin gene and negative control vector were transfected into human umbilical vein endothelial cells(HUVEC) incubated with 50 ng/mL of recombinant VEGF in vitro by lipofectamine 2000 .Transfection after 48 h ,the expression of survivin mRNA and protein was detected by quantitative real‐time PCR and Western blot ,respectively .HUVEC proliferation was assayed by four methylthiazolyl tetrazolium(MTT) and cell apoptosis was detected by TUNEL .Results (1)Transfection with survivin‐shR‐NA vector significantly down‐regulated the expression of survivin mRNA and protein as compared with the control group ,after transfection of 48 h(P<0 .05) .(2)After survivin‐shRNA vector transfected ,the proliferation of HUVEC decreased significantly . After transfection 24 ,48 ,72 h ,the growth inhibition rate were (13 .53 ± 3 .91)% ,(38 .97 ± 1 .82)% ,(65 .75 ± 1 .83)% respective‐ly ,at 72 hours after transfection was the most significant .(3)The apoptosis rate of experimental group was (28 .07 ± 1 .71)% , which was higher than the negative control group (11 .45 ± 1 .52)% and blank control group (10 .04 ± 1 .46)% (P<0 .05) .Conclu‐sion The shRNA‐mediated mediated silencing of the survivin‐gene could significantly inhibit proliferation and promote the apopto‐sis of rheumatoid arthritis synovial fibroblasts by regulating survivin expression .

In vitro study of BRD4 inhibitor GSK525762A against primary adult common B-cell acute lymphoblastic leukemia cells in vitro / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the effects of bromodomain-containing protein 4 (BRD4) inhibitor GSK525762A on the proliferation and apoptosis of primary common B-cell acute lymphoblastic leukemia (common B-ALL) cells from adult patients, then to further explore the possible mechanisms.</p><p><b>METHODS</b>Purified leukemia cells from 14 common B-ALL adult patients (4 Ph⁺ and 10 Ph⁻ cases) were obtained by flow cytometry sorting, and maintained in a mimic bone marrow microenvironment culture system for short-term culture. Leukemia cells were treated with various concentrations of GSK525762A. The inhibitory effects of BRD4 inhibitor on common B-ALL leukemia cells were measured by CCK-8 assay and the apoptosis of those cells was determined by AnnexinⅤ/7-AAD staining using flow cytometry. The transcripts of c-MYC, CDK6 and Bcl-2 were detected by quantitative RT-PCR, and the expression of c-MYC, CDK6 and Bcl-2 proteins were detected via Western blot.</p><p><b>RESULTS</b>GSK525762A could inhibit the proliferation of leukemia cells from all 14 common B-ALL patients in a dose-dependent manner, the median value of IC50 was 256.25 (90.64-1 378.39)nmol/L. GSK525762A could promote cells apoptosis of B-ALL leukemia cells in a dose-dependent manner, the median apoptosis rates respectively were 45.17%(9.38%-70.91%), 66.02% (24.36%-96.34%) and 89.29% (39.29%-99.37%) after treated by 500, 1 000 and 2 500 nmol/L GSK525762A. GSK525762A has a similar effect on Ph⁺ ALL and Ph⁻ B-ALL, but the effect of proliferation inhibition and apoptosis enhancement on Ph+ B-ALL is weaker than that on Ph⁻ B-ALL. Compared with vehicle control group, the levels of c-MYC, Bcl-2 and CDK6 transcripts in leukemic cells were reduced after treatment for 24 h and 48 h by 1 000 nmol/L GSK525762A, and there are no significant differences in the downregulation of c-MYC and CDK6 mRNA between Ph⁺ and Ph⁻ B-ALL; however, the inhibitory effect on Bcl-2 transcription was weaker in Ph⁺ B-ALL cells than that in Ph⁻ B-ALL cells. Moreover, c-MYC, Bcl-2 and CDK6 protein levels decreased in GSK525762A treated group.</p><p><b>CONCLUSION</b>GSK525762A could strongly inhibit the proliferation of common B-ALL and trigger apoptosis; meanwhile it has certain effects against Ph⁺ ALL in vitro. The effect may be achieved by down-regulation of c-MYC, CDK6 and Bcl-2 expression.</p>

Role of cerebrospinal fluid cytology detection on meningeal cancer / 中国综合临床

Objective To investigate the role of cerebrospinal fluie( CSF)cytology on eiagnosis of meningeal cancer. Methods Retrospectively analyzee the clinical eata of 23 cases with meningeal cancer. Results (1)Of 23 patients,CSF eetection of 17 cases were showee with cancer cells at the first lumbar puncture,3 cases at the 2ne lumbar puncture,2 cases at the 3re eetection,ane 1 case at 4th eetection.(2)Of 23 cases with cerebrospinal fluie cytology positive patients,17 cases were carriee cranial MRI scan. The MRI showee that 9 were normal ane 8 cases were brain parenchyma ane meningeal image enhance. Ten cases were performee the enhancee MRI scan,5 cases were with extensive meningeal thickening,3 cases were showee meningeal extensive enhancement of brain surfer ane intracranial cerebral sulci,1 case was the circular shaeow strengthen at eifferent size at next to the eouble lateral parietal,occipital ane left cerebella hemisphere,ans 1 case was with tough brain surface ane abnormally long T1,long T2 signal at lateral ventricles,thire ventricle, fourth ventricle epeneymal.(3)The relationship between the primary tumor ane cancer eetection in cerebrospinal fluie:19 patients were foune with primary tumor lesions,inclueing 5 cases leukemia(3 cases with acute lymphoblastic leukemia,2 cases with acute non-lymphocytic leukemia),4 cases with lung cancer,3 cases with breast cancer,2 cases with brain lymphoma,1 case with melanoma,1 case with Hoegkin′s lymphoma,l case with nasopharyngeal carcinoma, 1 case with vaginal squamous cell carcinoma, 1 case with gastric carcinoma. Conclusion Multiple cerebrospinal fluie cytology eetection may improve meningeal cancer eetection rate. CSF cytology eetection can improve eiagnosis rate of meningeal cancer. No relationship between the eetection of cancer cells in cerebrospinal fluie ane brain MRI meningeal lesions,ane the further research neee to be eone with expaneing the sample size.

Analysis of health inspection and related professional settings in colleges of China / 中华医学教育探索杂志

Objective To study the cultivation status of health inspection and the setting of the related major of this subject in colleges in China.Methods The setting situation of health inspection in undergraduate and junior colleges were searched from the professional information database of higher vocational schools,information query system of Chinese university and official websites of 7 junior colleges and 167 undergraduate colleges.Excel 2007 was used for inputting and sorting out data,and the data were combed and analyzed.Results The setting of college level of health inspection was approved in 2004,and there were 6 record colleges in 2014.The undergraduate level of related major was set in 2012 and only 1 college recruited students.Among 167 undergraduate universities,only 2 universities carried out professional education of health inspection,74 universities carried out professional education of health law and 65 universities carried out professional education of health management.Conclusion The subject of health inspection has been set in undergraduate and junior colleges for a short time and there are a few admissions; Professional education of preventive medicine,health law,health management and health inspection are relatively extensive.