Objective To investigate the antitumor effect of special promoter-controlled Gibbon ape leukemia virus membrane fusion glycoprotein (GALV.fus) mediated by type Ⅰ herpes simplex virus (HSV-Ⅰ) on lung adenocarcinoma. Methods Recombinant HSV-Ⅰ plasmids encoding GALV.fus was introduced into green monkey kidney cells(Vero)by liposome to amplify the virus, and then the virus was transfected into lung adenocarcinoma (A549), human fetal fibroblasts (HFL-Ⅰ GNHu 5) and human lung adenocarcinoma xenografts which were established in nude mice subcutaneously to observe antitumor and cytotoxic effect in vitro and in vivo; Recombined cytomegalovirus (CMV) containing GALV.fus or enhanced green fluorescence protein were served as control. Results Recombinant HSV-Ⅰ virus were packed successfully. Heterotransplantative tumourigenicity of the tumour was 100% in nude mice after A549 cells were inoculated. Recombinant HSV-Ⅰvirus exerted obvious antitumor effects in vitro, with relative survival rate of 23%, while for CMV virus containing GALV. fus, the rate was 20%, and for CMV virus encoding EGFP, the rate was 68%. Recombinant HSV-Ⅰvirus also showed striking antitumor effect on the implanted tumor. Conclusion GALV.fus has powerful effect against lung cancer in vitro and in vivo and maybe a promising candidate for gene therapy.

Objective:To explore the efficient construction of HSF1 gene knockout mouse model using CRISPR/Cas9 gene editing technology, and to establish the early basis for the mouse model of primary osteosarcoma.Methods:According to exon 9 of HSF1 gene structure, the corresponding GRNA (guideRNA) was selected and screened. Then the transcription template of sgRNA (small guide RNA) was amplified by PCR, and four up stream primers were obtained. Subsequently, sgRNA was transcribed in vitro and screened by Tube Screen platform to screen the sgRNA with effective cutting, and the sgRNA with the highest cutting efficiency was selected from the screening results for subsequent experiments. The transcription template of SPCas9mRNA was amplified by PCR, and then Cas9mRNA was transcribed in vitro. The sgRNA transcribed in vitro and Cas9mRNA were injected into the fertilized eggs of healthy C57BL/6 mice, and the tissue was extracted from the tail of the born mice and identified by PCR sequencing. Heterozygous female mice of F0 generation were selected to mate with wild-type male mice too btain F1 generation off spring. The mutation of gene bases of F1 generation mice was detected by AGAR gel electrophoresis and gene sequencing. The heterozygous male mice of the F1 generation and female mice of the F0 generation were back crossed to obtain the F2 generation daughter mice. The tail tissues were cut and sequenced to obtain the F2 generation homozygous knockout mice. PCR was used to observe the cutting efficiency of sgRNA and the sequencing of rat tail tissue, and SNAPGene software was used for gene sequence alignment to determine the deletion of base fragments.Results:The up stream primers sgRNA-1 Primer-f, sgRNA-2 Primer-f, sgRNA-3 Primer-f, sgRNA-4 Primer-f and down stream primers sgRNA-4 Primer -r were obtained by PCR amplification. After in vitro tran scription and screening of sgrRNA, sgrRNA-1, sgrRNA-2 and sgrRNA-4 had high cleavage efficiency and were selected for subsequent experiments. T7 promoter was added to the 5 'end of Cas9 mRNA, and Cas9 mRNA was obtained by PCR and in vitro transcription kit. Mixed Cas9-sgRNA solution was injected into the fertilized eggs of mice and cultured. The cultured two-cell fertilized eggs were injected into the ampulla of the pseudo pregnant female mice, and the F0 generation mice were obtained successfully. A total of 8 heterozygous mice of F0 generation were obtained by Agar gel electrophoresis. Three heterozygous knockout mice of F1 generation were obtained by breeding the female heterozygous mice of F0 generation with healthy wild-type male mice and PCR and sequencing. Three heterozygous male mice of F1 generation were back crossed with female mice of F0 generation 3 to obtain F2 generation mice. Through the observation of electrophoresis and sequencing results of F2 generation mice, it was confirmed that 7 mice were missing HSF1 base sequence, and the electrophoresis results showed mutant bands and no wild-type bands, which were identified as homozygous. The F2 generation homozygous mice were able to breed stably. As eries of results proved that the HSF1 gene knockout mouse model was successfully established in this experiment.Conclusion:CRISPR/Cas9 technology was successfully used to construct HSF1 gene knockout mouse model, with strong stability and high reproducibility, which laida foundation for further study of HSF1 gene expression products and establishment of mouse model of primary osteosarcoma.

Objective To investigate procedure and therapeutic effect of resection and reconstruction for axis tumors through the sub mandible approach. Methods Between December 2004 to June 2010,17 patients with axis neoplasm underwent tumor resection and antero-posterior reconstruction through the combined the sub mandible-inner sternocleidomastoid muscle (SMIS) approach and posterior approach. Tumor lesions involved C2 in 11 cases, C2-3 in 4, C2-4 in 2. Eight cases involved vertebral body, and 9 involved both vertebral body and element. Fourteen primary lesions including 4 giant cell tumors, 4 plasmocytomas, 2 chordomas, 2 eosinophilic granulomas, 1 hemangiopericytomas and 1 lymphoma, and 3 metastatic lesions were involved in this study. Three types of reconstruction in upper cervical spine including titanium mesh plus vertically placed titanium plate, titanium mesh plus obliquely placed titanium plate and trimmed titanium mesh alone, were adopted after anterior tumor resection, and then posterior tumor resection and reconstruction were performed. Results All patients experienced pain relief and neurological improvement after surgery. Except for one incidence of screw pull-out which was corrected by a revision surgery, solid fusion was achieved in all patients. A follow-up period of 6 months to 6 years was available for this study. One patient died of cerebral infarction 9 months postoperative. Two patients with chordoma relapsed 13 months and18 months postoperative, respectively, of whom one died of high plegia and respiratory failure, and the other was alive with disease. Two patients with metastasis died of multiple remote metastases 12 months and 18 months postoperative, respectively. Conclusion Through the SMIS apporach, a satisfactory exposure can be obtained for axis tumor resection and reconstruction. Anterior reconstruction of upper cervical spine after tumor resection can be achieved with internal fixation system of cervical spine, which can improve intraopera-tive safety. The combined anterior reconstruction and posterior occipito-cervcial fixation can provide immediate stability, and benefit maintaining stability of upper cervical spine.

Objective:To compare the clinical characteristics of congenital chylothorax in preterm and term infants.Method:From January 2011 to December 2019, the clinical data of infants with congenital chylothorax admitted to our hospital were retrospectively analyzed. The infants were assigned into preterm group (<37 weeks) and term group (≥37 weeks) according to their gestational age. The general information, clinical manifestations, laboratory results, treatment and prognosis of the two groups were compared.Result:A total of 34 infants with congenital chylothorax were included, including 11 premature infants and 23 term infants. No significant differences existed in gender, delivery mode, prenatal diagnosis of pleural effusion, congenital heart disease/chromosome abnormality, birth asphyxia, dyspnea, fetal edema, and location of effusion between the two groups ( P>0.05). Compared with term group, preterm group had significantly fewer leukocytes [3 245(1 007, 7 403)×10 6/L vs. 10 214(6 233,16 458)×10 6/L] and lower protein level [(28.1±7.6) g/L vs. (33.3±6.3) g/L] in the pleural fluid ( P<0.05). No significant differences existed in the proportion of pleural lymphocytes between the two groups ( P>0.05). The proportion of mechanical ventilation (MV) in the preterm group was statistically higher than that the term group [100%(11/11) vs. 65.2%(15/23)], and the duration of MV was statistically longer than the term group [(16(10,25) d) vs. (1(0,11) d)] ( P<0.05). No significant differences existed between the two groups in the application of other treatment options (thoracentesis/drainage, fasting, octreotide and erythromycin pleural injection), time needed for the disappearance of effusion, duration of hospital stay and cure/improvement rate ( P>0.05). Conclusion:Preterm infants may have lower leukocyte count and protein level in the pleural effusion than the term infants. Both preterm and term infants have higher proportion of lymphocytes in the pleural effusion fluid. Although most preterm infants need ventilator support after delivery, most of them achieve complete remission after conservative treatment and the overall prognosis is as good as term infants.

AIM:Using bioinformatics analysis methods to identify the hub genes involved in myocardial isch-emia-reperfusion injury(MIRI).METHODS:Firstly,the rat MIRI related dataset GSE122020,E-MEXP-2098,and E-GEOD-4105 were downloaded from the database.Secondly,differentially expressed genes(DEGs)were screened from each dataset using the linear models for microarray data(limma)package,and robust DEGs were filtered using the robust rank aggregation(RRA)method.In addition,the surrogate variable analysis(SVA)package was used to merge all datas-ets into one,and merged DEGs were screened using the limma package.The common DEGs were obtained by taking the intersection of the two channels of DEGs.Next,the protein-protein interaction(PPI)network of common DEGs was con-structed,and the hub genes were identified using the density-maximizing neighborhood component(DMNC)algorithm.The receiver operating characteristic curve(ROC)was plotted to evaluate the diagnostic performance of the hub gene.Then,the mRNA and protein expression levels of hub genes were detected in the rat MIRI model,and the literature re-view analysis was carried out on the involvement of hub genes in MIRI.Finally,the gene set enrichment analysis(GSEA)was performed on hub gene to further reveal the possible mechanism in mediating MIRI.RESULTS:A total of 143 robust DEGs and 48 merged DEGs were identified.After taking the intersection of the two,48 common DEGs were obtained.In the PPI network of common DEGs,5 hub genes were screened out,namely MYC proto-oncogene bHLH transcription fac-tor(MYC),prostaglandin-endoperoxide synthase 2(PTGS2),heme oxygenase 1(HMOX1),caspase-3(CASP3),and plasminogen activator urokinase receptor(PLAUR).The ROC results showed that the area under the curve values for all hub genes were greater than 0.8.MYC,PTGS2,CASP3,and PLAUR showed high mRNA and protein expression in rat MIRI,while there was no difference in mRNA and protein expression for HMOX1.The literature review revealed that among the 5 hub genes,only PLAUR has not been reported to be involved in MIRI.The GSEA results for PLAUR indicat-ed that its functional enrichment mainly focused on pathways such as NOD-like receptor signaling pathway,P53 signaling pathway,Toll-like receptor signaling pathway,apoptosis,and fatty acid metabolism.CONCLUSION:MYC,PTGS2,CASP3,HMOX1,and PLAUR are involved in the pathological process of MIRI.PLAUR is a potential hub gene that can mediate MIRI by regulating pathways such as NOD like receptor signaling,P53 signaling,Toll like receptor signaling,cell apoptosis,and fatty acid metabolism.The results can provide reference for further investigation into the molecular mechanisms and therapeutic targets of MIRI.

Background: Symptoms of irritable bowel syndrome with diarrhea (IBS-D) are known to be influenced by circadian oscillation; however, the pathophysiological mechanism is still unclear. Aims: To investigate the role and underlying mechanism of colon circadian clock gene Bmal1 involved in the occurrence of symptoms in IBS-D patients. Methods: Forty-six patients with IBS-D and 34 normal controls from Army Medical Center of PLA during September 2018 to February 2021 were recruited in this study. IBS-severity scoring system (IBS-SSS) was used to evaluate the severity of IBS-D symptoms. A colonoscopy was performed to obtain biopsy specimens from rectosigmoid colon. The concentration of 5-hydroxytryptamine (5-HT), and expressions of Bmal1 and chromogranin A (CgA), a biomarker of enterochromaffin cells (EC cells), in colonic mucosa were detected by ELISA and double-labeled immunofluorescence, respectively. Results: Both the 5-HT concentration and number of EC cells in colonic mucosa of IBS-D patients were significantly higher than those of the normal controls (all P< 0.05). Bmal1 was mainly expressed in intestinal epithelial cells and was highly expressed in EC cells. Co-expression of Bmal1 and CgA was observed. Compared with the normal control group biopsied at the same time point, expression of Bmal1 was significantly higher in specimens taken at 9 a.m., and expression of Bmal1 was significantly lower in specimens taken at 17 p.m. in IBS-D patients (all P< 0.05). Spearman correlation coefficient analysis showed that Bmal1 expression at 9 a.m. was positively correlated with the total score of IBS-SSS and subscore of abdominal pain and discomfort (r

ObjectiveTo evaluate the efficacy of TCM health management based on tertiary management system for patients with myocardial infarction （MI）. MethodsA total of 277 patients with non-acute MI were enrolled and given comprehensive TCM health management strategies including health education， lifestyle adjustment， risk factors control， medication and TCM appropriate techniques management through a tertiary management system with "the patient as the core， village/community physicians as the main executive body， and tertiary TCM hospital specialists as the leading body"， for a period of 12 months by using a prospective single-arm cohort study. Through patient reporting and medical records surveys， various indicators before and after 12-month management were collected and compared. The primary efficacy indicators were readmission rate and recurrent exacerbation rate， and the secondary efficacy indicators included disease awareness-related indicators， lifestyle behavior-related indicators， cardiovascular risk factor-related indicators and Canadian cardiovascular society （CCS） cardiac function classification. ResultsA total of 255 patients completed the study and were included in the final analysis. The recurrent exacerbation and readmission rates of patients after management were 23.14% （59 cases） and 20.25% （49 cases）， respectively， significantly lower than 36.08% （92 cases） and 53.72% （130 cases） before management （P＜0.05）. Except for knowledge on diabetes diagnostic criteria with no significant difference before and after management （P＞0.05）， awareness of other knowledge with regard to the prevention and treatment of cardiovascular and cerebrovascular diseases were improved after management （P＜0.01）， as well as the total score （P＜0.01）. In terms of daily life behaviors， the rates of salty diet， sweet diet and greasy diet were significantly lower than baseline， while the rate of moderate exercise was significantly higher （P＜0.05 or P＜0.01）； the rates of ongoing smoking and vigorous exercise were not significantly changed （P＞0.05）. For cardiovascular risk factors， patients' total cholesterol， low-density lipoprotein （LDL） cholesterol， triglycerides， fasting glucose， total depression assessment scale score， and total anxiety assessment scale score were significantly reduced after management （P＜0.01）. Systolic blood pressure and body mass index （BMI） were both higher after management （P＜0.05 or P＜0.01）， and there was no statistically significant difference in diastolic blood pressure （P＞0.05）. In terms of the cardiovascular disease risk factors reaching the standard levels， the rate of LDL cholesterol ＜1.8 mmol/L significantly increased （P＜0.01）， while the rate of BMI ＜24 kg/m² and the rate of systolic blood pressure ＜140 mmHg both decreased significantly （P＜0.05 or P＜0.01） from baseline； the diastolic blood pressure and rate of fasting glucose ＜7.0 mmol/L were not significantly changed （P＞0.05）. The patients' CCS cardiac function classification was significantly reduced （P＜0.05）. ConclusionTCM health management based on the tertiary management system can enhance MI patients' awareness of the disease， change poor lifestyle habits， reduce risk factors such as blood lipids and blood glucose， improve anxiety and depression， increase activity tolerance， and reduce their recurrence exacerbation and readmission rates， which is worthy of clinical promotion.

Objective To study the characteristics of noise hazards in automobile sand casting industry and understand the impact of noise on the health of automobile foundry workers, and to provide scientific data support for the control of noise hazards. Methods An automobile sand casting enterprise was selected as the research object. The post noise intensity test results from 2010 to 2019, the noise spectrum test results of each post in 2019, and the prevalence of occupational noise deafness in the 10 years from 2010 to 2019 were analyzed and sorted out. Using chi-square test and t-test the noise hazard characteristics and noise intensity changes were statistically analyzed. Results 1. The noise LEX.8h exceeding standard rate of foundry enterprises in the 10 years from 2010 to 2019 was not statistically significant（χ²=6.76, P=0.66), and the mean value of noise LEX.8h fluctuated horizontally in the 10 years, which was basically stable between 80 - 90 dB (A). 2. The mean value of noise intensity of sand falling, sand preparation, cleaning and shot blasting in the first five years was significantly different compared to that in the last five years. There was no statistically significant difference in other posts. 3. Among all posts in the foundry sand casting industry, the noise spectrum of core-making and cleaning posts was mainly medium and high frequency, while the noise spectrum of other posts was mainly medium and low frequency. 4, Cases of occupational noise deafness were mainly concentrated in cleaning and polishing workers, melting chemical workers, inspectors and other types of workers. Conclusions The noise hazards in the automobile foundry sand casting industry are mainly concentrated on the posts of molding, and sand falling, and the noise intensity and the over standard rate remain high in the past 10 years. The noise of each post is mainly medium and low frequency, and the noise of a few individual posts is mainly medium and high frequency. In recent years, due to the automatic renewal of equipment, the noise intensity of some posts such as sand dropping, sand mixing, cleaning and shot blasting has decreased, but the overall noise exceeding standard rate has basically not changed, and the noise hazard is still serious. The noise hazard of foundry sand casting enterprises is difficult to be controlled, and the risk of hearing loss of foundry workers is still high.