OBJECTIVE:To establish the method for the detection of the known glucagon-like peptide 1 receptor (GLP1R) gene mutation site rs3765467(NT_007592.16,position:39065819),and to evaluate its accuracy and practicability. METHODS:Peripheral venous blood samples of 72 healthy subjects were collected in medical examination center of our hospital during Oct. 2015-Feb. 2016. The whole blood DNA was extracted by column extraction method. After amplified by touch down PCR,high reso-lution melting(HRM)method was adopted to analyze amplified product. Sequencing verification by double stranded chain termina-tion method(Sanger sequencing method)was performed for 38 test samples. The results of 2 methods were compared. RESULTS:The results of mutation scanning showed that there were 39065817 and 39065819 polymorphism sites in amplified segments. Four types of mutations were detected by HRM method [GCG/GCG,GCA/GCG or ACG/GCG,GCA/GCA or ACG/ACG,A(G) CA(G)],but 6 types of mutations was detected by Sanger sequencing method [GCG/GCG,ACG/GCG,ACG/ACG,A(G)CA (G),GCA/GCG,GCA/GCA]. CONCLUSIONS:HRM method can identify GCG/GCG and A(G)CA(G)genotype,but can not identify GCA/GCG and ACG/GCG heterozygous mutation,GCA/GCA and ACG/ACG homozygous mutation. The method is not suitable for the detection of single nucleotide polymorphism for multiple neighboring sites. In the detection of single nucleotide mu-tation,economical and simple method should be selected after comprehensive analysis of sequence.

Objective To discuss the method, safety and selection of instrument in bone biopsy under digital radiography-guided. Methods Bone biopsy in 41 patients with bone lesion were performed under digital radiography-guided. Using the MDTECH super-core Ⅱ biopsy instrument, the MDTECH biopsy needle and osseous drill.Results The successful rate was 100% and the posilive rate was90.3%. None of them had severe complications. Conclusion Bone biopsy is a safe , accurate and effective method under digital radiography-guided.

Vaccination is the primary strategy for the prevention and control of pandemic influenza. Because influenza virus is highly variable across strains, universal influenza vaccines need to be developed to address this problem. This review describes the research progress in conserved epitopes of influenza virus, the advances in the research and development of universal influenza vaccines based on the relatively conserved sequences of NP, M2e, HA2, and headless HA, the mechanisms of cross-protection, and the methods to improve cross-protection.
Animals ; Cross Reactions ; Humans ; Orthomyxoviridae ; immunology ; Species Specificity ; Viral Proteins ; immunology ; Viral Vaccines ; genetics ; immunology

This article gives a profound exploration on the theoretical origin, prohibitions and mechanisms of moxibustion for heat syndrome. Based upon the ancient and modem literature, this article also gives a classified summarization on diseases of moxibustion for heat syndrome, in order to obtain a thorough understanding about the theory and clinical application of moxibustion for heat syndrome, hence to further perfect the theory of moxibustion and guide the clinical practice.

AIM:To investigate the effect of angiotensin II type 1 receptor (AT1R)-calcineurin (CaN) signa-ling pathway on the expression of sodium current channel Nav 1.5 at mRNA and protein levels in the hypertrophic ventricu-lar myocytes from neonatal rats .METHODS:The ventricular myocytes were isolated from the ventricles of 1-day-old neo-natal Sprague-Dawley rats and were divided into 4 groups according to different drug intervention as control group , pheny-lephrine (PE) group, losartan (Los)+PE group and cyclosporin A (CsA)+PE group.The method of RNA interference mediated by adenovirus carrying short hairpin RNA ( shRNA) was used to knock down the gene which encodes the beta subtype of CaN A subunit (CnAβ) and the cells were divided into 4 groups as Ad-Null group, Ad-Null+PE group, Ad-CnAβshRNA1 group and Ad-CnAβshRNA1+PE group.The mRNA expression of brain natriuretic peptide ( BNP) ,β-my-osin heavy chain (β-MHC) and Nav1.5 was detected by RT-qPCR.The protein levels of CnAβand Nav1.5 in the whole-cell extracts were determined by Western blot analysis .RESULTS:Treatment of the neonatal rat ventricular myocytes with PE for 24 h increased the protein-to-DNA ratio and the mRNA expression of BNP and β-MHC.The size of the cell surface was also increased after PE treatment .Treatment of the cells with PE increased the protein expression of CnAβ, and re-duced the protein expression of Nav 1.5.Both Los and CsA prevented those effects of PE .The mRNA expression of Nav1.5 was reduced by PE , and no significant difference of Nav 1.5 mRNA expression among PE group , Los+PE group and CsA+PE group was observed .Silencing of CnAβin the neonatal rat ventricular myocytes using Ad-CnAβshRNA1 inhibited the ability of PE to increase the mRNA expression of BNP , and diminished the ability of PE to reduce the protein expression of Nav1.5.CONCLUSION:AT1 R-CaN signaling pathway participates in regulating protein expression of Nav 1.5 in the hy-pertrophic ventricular myocytes from neonatal rats .

OBJECTIVE:To provide reference for formulating related provisions,guidelines and regulations for off-label drug use,standardizing medical behavior in medical health institutions. METHODS:According to the connotation of off-label drug use,Questionnaire for off-label drug use was developed for medical institutions,in which drug name,treatment disease,the type and use evidence ofoff-label drug usewere included. The questionnaires were delivered to pharmacists who participatedClinical phar-macist practice skills training in Sichuan province to collect the related information of off-label drug use for statistical analysis, and suggestions were put forward. RESULTS:Totally 150 questionnaires were sent out,124 were effectively received with effec-tively recovery of 82.7%;the surveyed pharmacists came from 22 medical institutions,including 18 tertiary hospitals and 4 second-ary hospitals;there were 128 information about off-label drug use,including 14 (10.9%) with incomplete reporting information. The another 114 information were major in“super indication”(61) and“super administration”(43),accounting for 53.5% and 37.7%,respectively,and there are 8 other drug overdose(7.0%),2 super object(1.8%). The relevant off-label drug use mainly included anti-infection drugs (34 information,29.8%),anti-tumor drugs (16 information,14.0%),TCM injections (8 informa-tion,7.0%),and etc.;the relative concentration of varieties were metronidazole,dexamethasone,gentamicin,methotrexate, azithromycin,irinotecan,vancomycin,nystatin,etc.. Some medical institutions were absence of effective regulatory measures. CONCLUSIONS:Off-label drug use is quite common in medical institutions of Sichuan province,some of them exist obvious med-ication risk. It is suggested that the state should be as soon as possible to develop related provisions,guidelines and regulations for off-label drug use,endowing legal force for off-label drug use;medical staff should avoid no evidence-based drug use,unreason-able or even unnecessary off-label drug use;pharmaceutical production enterprises should strengthen communication with medical institutions and pay attention to collect information related to the clinical medication,timely update the drug instructions to ensure the legitimate rights for patients and the interests for medical staff.

Objective To investigate does intracellular protein degradation pathway play an important role in decrease of endothelial nitric oxide synthase (eNOS) in human umbilical vein endothelial cells (HUVECs).MethodsTo establish a primary HUVECs culture methods,the HUVECs were incubated with concentration gradient group of TNF-α(0.01,0.1,1 and 10 ng/mL) in different time periods (24,48 and 72 h).The HUVECs were pretreated with NH4Cl or treated with caspase inhibitor or MG-132 1.5 h prior to incubation for an additional 24 h with TNF-α.The expression of eNOS was detected via Western blot assay.Results Treatment of the HUVECs with TNF-α(0.01-10 ng/mL) led to a dose-dependent reduction of eNOS expression.And treatment with TNF-α(1 ng/mL) reduced the eNOS expression in a time-depended manner.Compared with the TNF-α group,the protein expression level of eNOS was obviously increased in the co-working group of MG-123 and TNF-α.Conclusions TNF-α induces degradation of eNOS through a ubiquitin-proteasome pathway.

Objective To assess the abstracts on randomized controlled trials ( RCT) in non-small cell lung cancer ( NSCLC) published in Chinese and their influencing factors.Methods RCT in NSCLC published in Chinese were included according to the CONSORT statement and their influencing factors were analyzed by RevMan 5.3 soft-ware.Results The titles were identified as random, randomization, blinding, statistical method, recruited partici-pants, trial registry and fund-supported, respectively, in 20%of the 2677 abstracts included in this study.Con-clusion The titles are identified as random, randomization, blinding, statistical method, recruited participants, trial registry and fund-supported in RCT published in Chinese.Although the abstracts are improved after the publication of CONSORT, they need to be further brushed up.

Objective: To explore the role of angiotensin receptor type I (AT1)-calcineurin (CaN) signaling pathway in transient outward potassium ion channel (Ito) remolding in hypertrophic atrial myocytes of neonatal rats. Methods: 1 day old neonatal rats’ atrial myocytes were isolated and the cells were divided into 4 groups:①Control group, normal cells were cultured for 24 h,②Stretching group, the cells were cultured for 24 h with mechanical stretching to induce hypertrophy,③Telmisartan group, the cells were treated by telmisartan at 1 μmol/L for 1 h, then cultured for 24 h and ④Cyclosporin-A (CsA) group, the cells were treated by CsA at 0.25 μg/ml for 1 h, then cultured for 24 h. The ratios of protein/DNA in myocytes were compared between Control group and Stretching group, cell hypertrophy was deifned by mRNA expression of atrial natriuretic peptide (ANP). Ito changes were detected by whole-cell patch clamping technique, proteins expressions of Kv4.3 and CaN A subunit were examined by Western blot analysis. Results: Compared with Control group, Stretching group showed obviously decreased Ito density and Kv4.3 protein expression, while increased CaN A protein expression; Compared with Stretching group, the above effects were reduced in Telmisartan group and CsA group. Conclusion: AT1-CaN signaling pathway was involved in the regulation of Ito channel remodeling in hypertrophic atrial myocytes of neonatal rats.

Abstract: Objective To analyze the spatial-temporal characteristics of the active pulmonary tuberculosis (PTB) and the pathogenic positive PTB in Fuling District of Chongqing during the 13th Five-Year Plan period, so as to explore the clustering areas, and provide scientific basis for the precise prevention and control of tuberculosis in Fuling District. Methods The PTB registration data of 27 townships in Fuling District from 2016 to 2020 were collected. The descriptive analysis were used to describe the temporal and spatial distribution characteristics of patients, SaTScan9.0 and ArcGis10.6 was used for spatial-temporal scanning analysis and local auto-correlation analysis. The results were visualized by ArcGis10.6. Results A total of 4 038 case of active PTB patients were registered and a downward trend was observed in PTB during the 13th Five-Year Plan period in Fuling District. The average annual registration rate of PTB was 70.17/100 000, and the annual PTB registration rate declined by 8.21%. The peak of active PTB and etiological positive PTB were mainly concentrated in March and June respectively. The top five streets of cumulative active PTB patients registered were Lizhi street, Dunren street, Chongyi street, Ma 'an street and Jiangdong street, accounting for 60.18% of the total registered PTB patients during the 13th Five-Year Plan period. The top three average annual registration rates were Dunren street (101.35/100 000), Chongyi street (101.34/100 000) and Wulingshan Township (99.21/100 000). The registered PTB from 2016 to 2020 showed a global auto-correlation (Moran's I=0.64, P<0.0001). The "high-high" area of active PTB and the etiological positive PTB all covered Lizhi street, Jiangdong street and Longqiao street. By scanning analysis of spatial-temporal, the primary cluster of active PTB concentrated in the main urban area south of the Yangtze River in Fuling during January 2016 to December 2017, and the primary cluster of pathogenic positive PTB concentrated in the main urban area south of the Yangtze River in Fuling and Jiangdong street during January 2019 to December 2020. Conclusions During the 13th Five-Year Plan period, there was the spatial-temporal clustering of PTB in Fuling District, which mainly gathered in the main urban area south of the Yangtze River in Fuling district and surrounding streets centered on Lizhi street.