OBJECTIVETo generate an HPV16 prophylactic vaccine candidate for cervical cancer.

METHODSHPV16 major capsid protein L1 gene and minor capsid protein L2 gene were amplified using PCR. These genes were mutated by PCR site-directed mutagenesis for removal of sequence motifs (TTTTTNT) which would cause transcription termination when expressed from a vaccinia virus early promoter, then inserted into a vaccinia virus expression vector. A strain replication-deficient recombinant vaccinia virus containing the mutant sequences was obtained through a homologous recombination and identified.

RESULTSThe nucleotide sequence remained the correct amino acid sequence of the L1 and L2 proteins after mutated. Full-length L1 and L2 proteins were generated in cells infected with the recombinant virus. The virus strain propagated at very low titer or could not reproduce in some kinds of cell derived from different human tissues.

CONCLUSIONSThe authors have generated a strain replication-deficient recombinant vaccinia virus expressing HPV16 L1 plus L2 proteins as an HPV16 prophylactic vaccine candidate for cervical cancer.

Capsid ; Capsid Proteins ; genetics ; Cell Line ; Cloning, Molecular ; Female ; Gene Expression ; Genetic Vectors ; Humans ; Oncogene Proteins, Viral ; genetics ; Papillomaviridae ; genetics ; physiology ; Papillomavirus Infections ; prevention & control ; Transfection ; Tumor Virus Infections ; prevention & control ; Uterine Cervical Neoplasms ; virology ; Vaccinia virus ; genetics ; physiology ; Virus Replication

Objective:To analyze the detection effect of novel coronavirus 2019 nucleic acid in throat swabs and nasal swabs of COVID-19 cases.Methods:Throat swabs and nasal swabs were simultaneously collected from 541 cases of suspected cases and close contacts. Real time PCR was used to detect novel coronavirus 2019 nucleic acid. The positive rate of nucleic acid and Ct values of ORF1ab gene and N gene were analyzed. Results:A total of 43 cases were confirmed positive by nucleic acid detection result ; 35 cases were positive and the positive rate was 6.47% in throat swabs, while 33 cases were positive and the positive rate was 6.10% in nasal swabs. There was no significant difference in the positive rate between the two types of samples ( χ2=0.06, P=0.80). The diagnostic consistency of the two specimens was good ( Kappa=0.72). Ct values of ORF1ab and N genes between the two groups were not significantly different, either. Conclusions:There was no significant difference in the detection of novel coronavirus 2019 between throat swabs and nasal swabs from COVID-19 patients by real-time PCR.

Objective To investigate the expressions of transforming growth factor β1 (TGF-β1) and brain natrium peptide (BNP) in patients with diastolic heart failure (DHF),and to explore the correlation between plasma levels of TGF-β1,BNP and TGF-β1/BNP with parameter of diastolic function,diastolic dysfunction and New York Heart Association (NYHA) classification of cardiac function.Methods Hospitalized patients with DHF from October 2016 to November 2017 in Beijing Chaoyang Hospital were selected as subjects.At the same time,the age-and gender-matched non-heart failure hospitalized patients were selected as the control.The diastolic function index (E/e') was measured using cardiac ultrasound spectral Doppler and tissue Doppler methods.The diastolic dysfunction classification was evaluated according to the American Society of Echocardiography guidelines.Cardiac function was evaluated with NYHA classification.The levels of plasma TGF-β1 and BNP were measured with method of enzyme linked immunosorbent assay (ELISA).The correlation between the indicators was analyzed and the receiver operating characteristic (ROC) curve was drawn.Results A total of 186 patients were enrolled,including 114 patients as DHF group [54 males and 60 females,mean age (70.75 ± 11.45) years old] and 72 cases as control group [41 males and 31 females,mean age (68.74 ± 10.86) years old].The levels of TGF-β1 [(77.68 ± 42.31) ng/L] and BNP [(1 153.84 ± 564.96) ng/L] in patients with DHF were significantly higher than those of the control group [(18.76 ± 13.70),(264.07 ± 179.43) ng/L,t =15.62,13.77,P ＜ 0.01].Pearson correlation analysis showed that level of plasma TGF-β1 had a significant liner correlation with index E/e' (r =0.582,P ＜ 0.01),level of plasma BNP had a low-degree liner correlation with index E/e' (r =0.261,P ＜ 0.01),and TGF-β1/BNP had no correlation with index E/e' (r =0.081,P ＞ 0.05).Spearman correlation analysis showed that the levels of TGF-β1 and BNP were significantly correlated with diastolic dysfunction grading (r =0.473,0.417,P ＜ 0.01),while TGF-β1/BNP had no correlation with diastolic dysfunction grading (r =0.062,P ＞ 0.05).Plasma TGF-β1 and BNP had low-degree correlation with NYHA classification of heart failure (r =0.309,0.326,P ＜ 0.01),TGF-β1/BNP had no correlation with NYHA classification of heart failure (r =0.011,P ＞ 0.05).Logistic analysis showed that both plasma TGF-β1 and BNP were independent predictors of DHF (OR =1.264,2.283,P ＜ 0.05 or ＜ 0.01).The area under ROC curve (AUC) of BNP for prediction of DHF was 0.937 ± 0.064,and TGF-β1 was 0.597 ± 0.042.AUC areas of BNP and TGF-β1 were significantly different (P ＜ 0.01).Conclusions The expressions of plasma TGF-β1 and BNP in patients with DHF are higher than those without DHF.The levels of plasma TGF-β1 and BNP are significantly correlated with index E/e',diastolic dysfunction grading and NYHA classification.Both elevated BNP and TGF-β1 levels are independent predictors of DHF.Both plasma BNP and TGF-β1 have auxiliary diagnostic value on DHF and the diagnostic value of plasma BNP is greater than plasma TGF-β1.

OBJECTIVE： To provide reference for standardizing rational use of set prescription preparation containing Coptis chinensis and its processed product. METHODS： By retrieving 2015 edition of Chinese Pharmacopeia·Guidelines for Clinical Drug Use（volume of TCM set prescription）， the inclusion of set prescription preparation containing C. chinensis and its processed product were summarized and analyzed. RESULTS&CONCLUSIONS： There were 127 set prescription preparations containing C. chinensis and its processed product included in 2015 edition of Chinese Pharmacopeia·Guidelines for Clinical Drug Use（volume of TCM set prescription）， among which， there were 83， 5， 2， 8， 4， 11， 6， 5 and 3 set prescription preparations for internal medicine， surgery， gynecology， pediatrics， dermatology， ophthalmology， pharyngology， stomatology， orthopedics and traumatology， respectively. There were 120 set prescription preparations containing C. chinensis（94.49％）， 2， 4 and 1 set prescription preparations containing prepared C. chinensis with vino， prepared C. chinensis with ginger and prepared C. chinensis with Euodia rutaecarpa， respectively. There were 39， 59 and 29 kinds of set prescription preparations with C. chinensis as main symptoms medicine， assists medicine and complication medicine， which were mainly for clearing heat， drying dampness， purging fire and detoxifying（104 kinds， 81.89％）. Main types included Shexiang niuhuang pills， Gegen cenlian tablets， Kaiwei jianpi pills， etc. By comparing the efficacy of C. chinensis and its processed product， it was found that the names of processed products were not standardized， the labeling of C. chinensis and its processed products were not uniform and the application was not standardized； the related contents still needed to be further improved. It is necessary to strengthen the research on the pharmacodynamic basis and processing standard of C. chinensis and carry out the investigation and correction of relevant publications in order to help improve the rational drug use level of set prescription preparations containing C. chinensis and its processed products.

Objective:To explore the effect of closed-loop nursing on self-efficacy and nursing satisfaction in children with nephrotic syndrome (NS) .Methods:From March 2019 to January 2021, convenience sampling was used to select 104 children with NS and their accompanying parents in Henan Provincial People's Hospital as the research object. The children were divided into the control group and the observation group by random number table method, 52 cases in each group. The control group received routine nursing, and the observation group carried out closed-loop nursing on the basis of the control group. The scores of the General Self-efficacy Scale and Satisfaction Scale were compared between the two groups.Results:After three months of intervention, the children's self-efficacy score in the observation group was higher than that in the control group, and the parents' satisfaction with nursing was 94.23% (49/52) , which was higher than 78.85% (41/52) in the control group. The differences were all statistically significant ( P<0.05) . Conclusions:The closed-loop nursing can improve the self-efficacy of children with NS and the satisfaction of their parents, which is worthy of clinical promotion.

Objective:To analyze the relationship between genotype and clinical phenotype of the MYH7-R453C mutation in five Chinese hypertrophic cardiomyopathy (HCM) families.Methods:A retrospective cohort study was conducted on 527 unrelated HCM probands who were first diagnosed at the First Affiliated Hospital of Air Force Medical University (Xijing Hospital) from February 2014 to July 2018, and the high-throughput whole exome targeted sequencing of 96 genes related to hereditary cardiovascular disease was performed on the probands. The probands carrying the MYH7-R453C mutation were screened out, and their family members carrying the mutation were verified using Sanger sequencing. Healthy individuals without family history of genetic diseases from the same period and ethnicity were recruited as controls. Clinical data such as echocardiography, 12-lead electrocardiogram, and cardiac magnetic resonance imaging of the probands and their family members were collected, and the correlation between patient genotype and clinical phenotype was analyzed. Endpoint or key events were recorded through hospital re-examination or telephone follow-up.Results:The MYH7-R453C mutation was detected in 5 HCM probands, and clinical data and genetic results of 20 family members, including probands, were collected. Among them, 13 carried the MYH7-R453C mutation, of which 12 were diagnosed with HCM, and one child (F1Ⅲ 5) experienced early changes of HCM. The seven family members who did not carry the MYH7-R453C mutation had normal echocardiograms and 12-lead electrocardiograms. Among the 12 patients diagnosed with HCM, 2 experienced (F2Ⅱ 7, F5Ⅰ 2) sudden cardiac death, 2 experienced (F1Ⅲ 1, F3Ⅲ 3) events of sudden cardiac death survival, 2(F1Ⅱ 2, F3Ⅱ 1) died from heart failure during the follow-up period. Combined with the initial visit and follow-up, 4 families (F1, F2, F3, F5) had a family history of sudden death, among which 3 families probands or multiple family members experiencing sudden death before the age of 30 and adverse outcomes such as implantation of implantable cardioverter-defibrillators after sudden death survival. Conclusions:In the five families with HCM carrying MYH7-R453C mutations, genotype is highly correlated with clinical phenotype, and patients have a high risk of sudden death and poor prognosis. Early diagnosis of individuals carrying the MYH7-R453C gene mutation, both within the patient′s family and in the patients themselves, is crucial for initiating early treatment, preventing sudden death, and assessing prognosis.

Objective To investigate the correlation between MEX3A and differentiation characteristics of gastric cancer and intestinal metaplasia,and its combination with caudal-related homeobox transcription factor 2(CDX2)and mucin 2(MUC2)and mucin 5AC(MUC5AC)to determine the role of carcinogenic intestinal metaplasia.Methods From January 2010 to December 2014,a total of 410 cases of gastric cancer and paracarcinoma paraffin-embedded tissue samples were selected from the Central Hospital Affiliated to Shenyang Medical College and the Second Hospital Affiliated to Shenyang Medical College.According to pathological diagnosis,they were divided into control group(mild superficial gastritis,79 cases),intestinal metaplasia group(149 cases)and gastric cancer group(182 cases).The expressions of MEX3A,CDX2,MUC2 and MUC5AC were detected by immunohistochemistry.Results MEX3A was highly expressed in gastric cancer group and intestinal metaplasia group,especially diffuse gastric cancer,poorly differentiated gastric cancer and type Ⅲ intestinal metaplasia(P<0.05).CDX2 and MUC2 were highly expressed in gastric cancer group and intestinal metaplasia group,especially intestinal type gastric cancer,highly and moderately differentiated gastric cancer,type Ⅰ and type Ⅱ intestinal metaplasia(P<0.05).The expression of MUC5AC was high in control group and low in gastric cancer group and intestinal metaplasia group,especially in intestinal type gastric cancer,type Ⅰ and type Ⅲ intestinal metaplasia(P<0.05).Gastric cancer and intestinal metaplasia differentiation were negatively correlated with MEX3A and MUC5AC expression,but positively correlated with CDX2 and MUC2 expression(P<0.05).MEX3A was negatively correlated with the expression of CDX2 and MUC2,and positively correlated with the expression of MUC5AC in gastric cancer(P<0.05).MEX3A was negatively correlated with the expression of CDX2 and MUC2 in intestinal metaplasia(P<0.05),while CDX2 was positively correlated with the expression of MUC2(P<0.05).Conclusion MEX3A is negatively correlated with gastric cancer and intestinal metaplasia differentiation.Gastric cancer is characterized by high MEX3A expression and low CDX2 and MUC2 expression.