ObjectiveTo investigate the effects of nicotine on the expression of Th1/Th2 cytokines and transcription factor T-bet/GATA-3 in cultured CD4+ T of rat sensitized by ovalbumin.Methods Two weeks after immunization by ovalbumin,splenic CD4+ T cells of Wistar rat were purified using CD4+T cell enrichment kit.Purified CD4+ T cells of rat were cultured and divided into 4 groups:a control group,1 μg/ml nicotine stimulated group,10 μg/ml nicotine stimulated group,100 μg/ml nicotine stimulated group.These cells,in their groups,were stimulated with or without nicotine and were all challenged simultaneously with OVA.Supernatants and cell pellets were harvested after being stimulated for 24 h.The concentration of IFN-γ and IL-4 in supernatants were measured by enzyme-linked immunosorbent assay (ELISA).Real-time PCR was used to determine the mRNA expression of T-bet and GATA-3 in CD4+T cells.Results ( 1 ) IFN-γproduction was significantly decreased in all nicotine treated groups [ ( 113.78±6.06) ng/L,(70.31±7.26) ng/L,(20.00±2.14) ng/L] compared with the control group[ (142.30± 5.89) ng/L],and the level of IL-4 was significantly increased in all nicotine treated groups [ (69.49±3.91) ng/L,(93.63±4.56) ng/L,(50.97±3.07) ng/L] compared with the control group[ (36.91±3.24) ng/L].(2) Expression of T-bet mRNA in all nicotine treated groups(0.73±0.03,0.57±0.04,0.31 ±0.00) was lower than that in the control group(0.98±0.09),but expression of GATA-3 mRNA in all nicotine treated groups (4.31±0.26,5.16±0.23,1.56±0.14) was significantly higher than that in the control group(1.00±0.07).Conclusion Nicotine may play a key role in the development of Th2-type allergic inflammation in asthma by promoting over-expression of GATA-3 mRNA and downregulating the expression of T-bet mRNA.

Objective To explore the mechanism of protective effect of liposome carried prostaglandin E 1 (PGE 1) on rat skin flap ischemia / reperfusin(I/R). Methods Island flaps of abdomen of adult SD rats were established for ischmia reperfusion injury models. Weighting, immunocytochemistry and in situ hybridization were employed. Results It was found that treatment with liposome carried PGE 1 significantly alleviated I/R induced muscular edema, attenuated neutrophil accumulation in the I/R tissue ( P

Studies on promoting thermogenesis of brown and beige adipose tissue have become a hot topic in various metabolic conditions, which based on the conclusions that brown and beige adipose tissue are able to facilitate weight loss and improve metabolic health.However, recent studies showed that there were several problems for the anti-obesity application via promoting brown/beige adipose tissue thermogenesis.In obese individuals, classical brown adipose tissue presents thermogenesis dysfunction.Moreover, the beige adipose tissue has significantly lower thermogenesis capability compared with brown adipose tissue.On such conditions, excessively excited sympathetic innervation is essential to increase energy consumption in obesity via increasing classic brown adipose thermogenesis and inducing white adipose tissue browning.However, excessive excited sympathetic nerve results in cardiovascular side effects.Additionally, excessive induction of white adipose tissue browning might disrupt the white adipose tissue homeostasis and aggravate the intrinsic metabolic disorders.Therefore, solving these practical application problems of brown/beige adipose tissue is a new research area for improving metabolic disorders.

Objective To observe the impact of hypoxia/reoxygenation on myocardial cytoskeletal proteins (α-actinin protein,tubulin protein,desmin protein) and to investigate EPO lessening the damage of myocardial cytoskeleton proteins in rats proved by culturing hypoxia/reoxygenation injured myocardial cells in presence of EPO.Methods The rat model of asphyxia-induced cardiac arrest was performed by turning-off the ventilator and clamping the endotracheal tube.After asphyxia for 8 minutes,CPR was carried out.A total of 24 rats were divided into normal group,ischemia/resuscitation (I/R) group and the EPO group (n =8).The model of myocardial dysfunction was determined 2 hours after restoration of spontaneous circulation (ROSC).The rats of EPO group were given EPO 5000 U/kg after ROSC.The rat heart specimens were collected.Actinin,Tubulin and Desmin protein were observed by SABC immunohistochemistry.The cultured cardiomyocytes were taken from neonatal rats and were divided into three groups:the normal group,the hypoxia/reoxygenation (H/R) group (hypoxia 10 h/reoxygenate 4h),the EPO group (hypoxia 10 h/reoxygenate 4 h,plus 10 U/mL EPO).The changes of tubulin and actinin in cultured cardiomyocytes were observe by Immunofluorescence.Results From immunohistochemistry,there were no significant difference in the optical density of actinin,tubulin and desmin among the normal,I/Rand EPO groups.After H/R injury,the structures of the actinin,tubulin protein were destroyed,the network structure of both protein were unclear in cultured myocardial cells.The grades of fluorescence intensity of actinin and tubulin in H/R group were significant lower than those in normal group,but there was no significant difference between H/R group and EPO group.Conclusions The damage of cytoskeleton during ischemia/reperfusion may be time-dependent.EPO has no beneficial effect on the cytoskeleton after I/R injury.

Objective To investigate the feasibility and effects of endoscopic surgical treatment of lumbar intervertebral disc herniation associated with veitebral osteochondrosis.Methods From June 2008 to December 2015,276 cases of lumbar intervertebral disc herniation associated with vertebral osteochondrosis were treated with endoscopic surgery,including 185 men and 91 women,with an average 39.2 years old (range,16-65 years old).The involved level included L2.3 in 2 cases,L3.4 in 9 cases,L4,5 in 126 cases and L5S1 in 139 cases.On preoperative axial CT,the diameter of ossification was more than half of the transverse or sagittal diameter of the spinal canal in 89 cases,and no more than half of the transverse and sagittal diameter of the spinal canal in 187 cases.All patients were operated on the side with serious symptom,181 cases were operated with mobile microendoscopic discectomy (MMED),and 95 cases were operated with percutaneous endoscopic surgery,including percutaneous transforaminal endoscopic discectomy (PTED) in 61 cases and the percutaneous interlaminar endoscopic discectomy (PIED) in 34 cases.The operation and complications were analyzed.Results The soft herniation,broken disc material and the periphery of compressing ossification were removed under the endoscope in all cases,until the nerve was well decompressed.However,the ossification was not complete resected.Dural sac tear occurred in 3 cases of MMED.In the early stage of PTED,2 cases converted to MMED because of intraoperative pain and difficulty,and one case had exiting nerve root injury.At the final follow-up of 12-60 months (average,20.6 months),visual analogue scale decreased from preoperative 8.5±1.2 to 1.0±0.9,Oswestry disability index decreased from preoperative 40.2±8.6 to 3.1±3.0.According to Macnab scale,the results were excellent in 89,good in 154 cases,moderate in 33 cases.Conclusion For most lumbar intervertebral disc herniation associated with vertebral osteochondrosis,good results can be achieve by removal of herniated and broken intervertebral disc and decompression of nerve with endoscope.Therefore,we speculate that the soft disc herniation and spinal stenosis are main pathogenic factors,and that the complete resection of ossification is not needed.

Objective:To investigate the effect of lipopolysaccharide (LPS) on primary neonatal rat cardiomyocytes (CMs) and human induced pluripotent stem cell-derived cardiomyocytes (hiPS-CMs).Methods:The hiPS-CMs and primary neonatal rat CMs were treated with different concentrations of LPS for 24 to 48 h. Then the cellular viability was analyzed by the xCELLigence RTCA Cardio system. The measurement of NPPB gene was studied by qRT-PCR and the gene expression analysis was performed by the qPCR array, in order to evaluate the cardiac inflammation effect induced by LPS.Results:The LPS exposure led to dysfunction in the primary neonatal rat CMs, which shown as an increase in beating rate and a decrease in contraction amplitude ( P<0.01), accompanied by an increased NPPB mRNA level ( P<0.01). There was no significant alteration in beating rate and the contraction amplitude in the corresponding concentration of the primary neonatal rat CMs ( P>0.05), as well as the NPPB mRNA level ( P>0.05). However, the expression of NPPB mRNA in hiPS-CMs was significantly different at a higher concentration of LPS (5 μg/mL~40 μg/mL) ( P<0.01), but the beating rate and the contraction amplitude showed no significant change, even the concentration of LPS up to 40 μg/mL ( P>0.05). Finally, the genes of C3, Gpnmb, Atf3, Il6r and Ly96 upregulated to 1.5 folds in the primary neonatal rat CMs. In comparison with primary neonatal rat CMs, the AK4, TOLLIP, SPP1, FABP1, IL6R, LY96 and C3 were over expression to 1.5 folds in the hiPS-CMs. Conclusions:In comparison with primary neonatal rat CMs, hiPS-CMs are markedly less injured by LPS and show a different pattern of inflammation gene expression.

Objective:To evaluate the value and efficacy of self-anchored anterior lumbar discectomy and fusion (SA-ALDF) for L 5 isthmic spondylolisthesis. Methods:From June 2018 to December 2019, a total of 11 cases of L 5 isthmic spondylolisthesis were treated with SA-ALDF, including 4 men and 7 women, aged 43.2±12.6 (range 29-63) years. All patients had intractable low back pain aggravating during standing activities and alleviating during rest, without lower extremity radicular symptoms. Imaging examination showed bilateral isthmus cleft of L 5 with spondylolisthesis of 1 degree in 10 cases and 2 degree in 1 case according to Meyerding grading system. Under general anesthesia and supine French position, transverse 6 cm incision was made. Then, the L 5S 1 intervertebral disc was exposed via extraperitoneal approach between the bifurcation of abdominal aorta and vena cava. The intervertebral disc was sufficiently removed. The intervertebral space was released and distracted followed by canal ventral decompression and sequential mold testing. Suitable self-anchoring cage filled with auto iliac cancellous bone was implanted to restore intervertebral height and lordosis as well as reduction of spondylolisthesis. Under fluoroscopic guidance, the distal anchoring plate was knocked into the sacrum followed by direct reduction and proximal anchoring plate locking in the L 5 vertebral body. The patients were followed up for 12.1±4.7 (range 6-18) months. The visual analogue score (VAS) and Oswestry dysfunction index (ODI) were evaluated. The reduction and fusion were evaluated on the X-ray films. Furthermore, the rate of spondylolisthesis, the height and the lordosis of intervertebral space were measured. Results:The operation was performed successfully in all the patients with operation duration 90±18 (range 70-120) min, intraoperative blood loss 30±16 (range 10-60) ml. No severe complication such as nerve and blood vessel injury occurred. All patients experienced alleviation of symptom during follow-up. X-rays confirmed that the spondylolisthesis and alignment were recovered obviously without obvious cage displacement. However, the loss of reduction was 63.2% for the grade 2 spondylolisthesis. At the final follow-up, VAS decreased from 6.1±2.1 to 0.9±0.5, ODI decreased from 43.6%±14.2% to 6.0%±3.4%. The spondylolisthesis recovered from 17.7%±10.3% to 8.0%±7.2% with reduction rate of 54.8%±21.6%. The interverbral height recovered from 6.4±2.1 mm to 9.8±3.9 mm and intervertebral lordosis recovered from 4.8°±2.9° to 9.6°±4.7°.Conclusion:SA-ALDF can provide satisfactory outcomes for selected L 5 isthmic spondylolisthesis of degree 1 without neurological compromise. However, its mechanical stability may be insufficient for isthmic spondylolisthesis of degree 2.

This study aims to learn drug resistance situation of four first-line anti TB drugs among 251 Mycobacterium tuberculosis isolates from Qinghai and to explore their relationships with genotypes by Spoligotyping,so as to provide basis for effective prevention of tuberculosis.Isolates of 251 M.tuberculosis were tested susceptibilities of four first-line drugs including isoniazid (INH),rifampicin (RFP),streptomycin(SM) and ethambutol (EMB) by using conventional proportion method and genotyped by Spoligotyping.Relationship between drug resistance and genotypes was analyzed statistically.Results showed the total drug resistance rate was 56.2％ (141/251) among 251 M.tuberculosis isolates.Resistance rates of four first-line drugs were 43.0％ (108/251) for INH,37.1％ (93/251) for RFP,39.0％ (98/251) for SM,27.9％ (70/251) for EMB respectively.Rate of multidrug-resistant TB (MDR TB) was 31.5％ (79/251).All 251 isolates of M.tuberculosis were typed by spoligotyping.The 185 (73.7 ％) were Beijing genotypes and 66 (26.3.％) were non-Beijing genotypes,and no statistical association was found with drug resistance.This paper concludes that isolates of M.tuberculosis prevail in Qinghai have both high rates of drug resistance and MDR and dominant isolates are Beijing genotypes by spoligotyping.

Objective:To analyze the sepsis related long non-coding RNA (lncRNA) and mRNA expression profiles based on Gene Expression Omnibus (GEO) datasets and bioinformatic analysis, and to analyze the sepsis-associated competing endogenous RNA (ceRNA) network based on microRNA (miRNA) database.Methods:The sepsis-related lncRNA dataset was downloaded from the GEO database, and the differential expression analysis was conducted by Bioconductor on the sepsis dataset to obtain differentially expressed lncRNA (DElncRNA) and differentially expressed mRNA (DEmRNA), and cluster heat map was drawn. miRNA combined with DElncRNA were predicted by miRcode. mRNA targeted by miRNA was simultaneously met by three databases: TargetScan, miRDB, and mirTarBase. The interaction relationship of lncRNA-miRNA-mRNA was obtained. The regulatory network visualization software CytoScape was used to draw ceRNA networks. DEmRNA in the ceRNA networks were imported into the Search Tool for the Retrieval of Interacting Genes Database (STRING) online database to draw the protein-protein interaction (PPI) map. The gene ontology (GO) function annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of DEmRNA were performed.Results:Dataset GSE89376 and GSE145227 were found from GEO database. Difference analysis showed there were 14 DElncRNA and 359 DEmRNA in the elderly group of GSE89376; 8 DElncRNA and 153 DEmRNA in the adult group of GSE89376; 1 232 DElncRNA and 1 224 DEmRNA in the children group of GSE145227. Clustering heatmap showed that there were significant differences in the expression of lncRNA and mRNA between the sepsis group and the control group. The ceRNA networks were constructed with miRNA. Several DElncRNA and multiple DEmRNA participated in the ceRNA network of sepsis. The PPI diagram demonstrated that several genes encoding proteins interacted with each other and form a multi-node interaction network with multiple genes encoding proteins. Functional annotation and enrichment analysis demonstrated that there might be a crosstalk mechanism on functionally related genes such as nuclear receptor activity, ligand-activated transcription factor activity, and steroid hormone receptor activity, and played a role in the occurrence and development of diseases through forkhead box transcription factor O (FoxO) signaling pathway, Janus kinase/signal transducers and activators of transcription (JAK/STAT) signaling pathway, p53 signaling pathway, and phosphateidylinositol 3-kinase (PI3K)/Akt signaling pathway.Conclusion:Through sepsis-related lncRNA-miRNA-mRNA ceRNA network and combining with KEGG pathway analysis, there were several lncRNA and mRNA participating in the ceRNA network related sepsis, which played an important role in several signal pathways.

Abstract: Objective To analyze the resistance and spatial distribution of Mycobacterium tuberculosis (MTB) to six commonly used anti-tuberculosis drugs in Qinghai Province from 2016 to 2019, so as to provide a reference for tuberculosis treatment and drug-resistant tuberculosis control. Methods A total of 1 182 identified strains of Mycobacterium tuberculosis in Qinghai Province from 2016 to 2019 were collected, and 6 anti-tuberculosis drugs were subjected to drug susceptibility tests and strain confirmed by the proportional method. By means of ArcMap10.7 and SaTScan10.1 software, map visualization, spatial autocorrelation analysis and spatial scanning of MTB drug resistance were performed to identify MTB drug resistance clusters in Qinghai Province. Results From 2016 to 2019, the total drug resistance (TDR) rate of 1 182 Mycobacterium tuberculosis strains in Qinghai Province was 23.77% (281/1 182), with a mono-resistance (MR) rate of 11.08% (131/1 182), a poly-resistance (PDR) rate of 3.89% (46/1 182), a multi-drug resistance (MDR) rate of 8.80% (104/1 182), and an extensive drug resistance (XDR) rate of 0.85% (10/1 182). The rates of MDR, XDR and TDR all showed a decreasing trend year by year (P<0.01). The drug resistance spectrum displayed 21 combinations. The TDR rate and MDR rate in the retreatment patients were higher than those of the initial treated patients, and the difference was statistically significant (χ2 TDR=22.784, χ2MDR=45.082, P<0.01). In terms of demographic characteristics, the TDR rate in males was higher than that in females, and the middle-aged group was higher than other age groups, and the differences were statistically significant (χ2=7.541, 10.825, P<0.05). The results of global spatial autocorrelation analysis showed that there was no statistical significance in the autocorrelation and obvious spatial clustering of MTB drug resistance in Qinghai Province from 2016 to 2019 (P>0.05), which indicated a random distribution. The results of spatiotemporal scanning showed that there was a kind of clustering area, but the clustering effect was not significant (P>0.05), indicating a random distribution. Conclusions The TDR of MTB in Qinghai Province from 2016 to 2019 showed a downward trend year by year. In comparison with the national average, the rate of multi-drug resistance and extensive drug resistance was still high, and most of the multi-drug resistance resulted from rifampicin and isoniazid. The drugresistant population mainly consisted of retreatment, males, and young and middle-aged pop