To express a gene of heat shock protein 70(HSP70) in E.coli , HSP70 gene was amplified by PCR. The PCR products were cloned into pUCm T vector and were sequenced; The HSP70 gene was subcloned into vector pET 21a(+) and expressed in E.coli. SDS PAGE and Western blot were employed to identify the expression of the HSP70 gene. Results showed that a fragment about 1 96kb was amplified by PCR. Sequence analysis revealed that the sequence of HSP70 was correct. The HSP70 gene was cloned into pET 21a(+) identified by enzyme digestion and PCR. SDS PAGE and Western blot showed that a M 72 000 protein was expressed and could be recognized by anti HSP70 antibody. Therefore,HSP70 gene has been successfully expressed in E.coli .

Objective To construct a fusion gene vaccine of gastric cancer MG7-Ag mimotope and heat shock protein 70, and to investigate the humoral and cellular immune response in mice induced by the vaccine. Methods The coding sequence of MG7 mimotope was incorporated with HSP70 gene at the 3' terminus by PCR amplification. Then the PCR product was cloned into pcDNA3.1(+) vector to construct the eukaryotic expression vector. The pcDNA3.1/MG7+hsp70 was sequenced to ensure the proper encoding. C57BL/6 mice were immunized with the fusion gene vaccine, and pcDNA3.1/hsp70 and empty pcDNA3.1 were used as controls. The serum was collected at the 3rd week after each immunization. Cellular ELISA was performed to evaluate the induced humoral immunity. The splenocytes were first separated at the 9th week for the assay of antigen specific CTL lysis activity by 51Cr release. Results A fragment about 2.0 kb was obtained by PCR amplification. Sequence analysis revealed that the sequence of mimotope was connected successfully to 3' terminus of hsp70 and the fusion gene was cloned into the pcDNA3.1(+) successfully. Cellular ELISA results suggested that the serum level of MG7-Ag antibody appeared in the vaccinated mice at 9th week, while no MG7-Ag antibody was detected in the controls. The results of the specific lysis rate of splenocytes showed no statistical difference between fusion gene vaccine group and control groups. Conclusion The fusion gene vaccine was constructed successfully and the specific humoral immune response was induced by the fusion gene vaccine.

ObjectiveTo explore the characteristics of the metabolite concentrations of the precentral gyrus in healthy Chinese adults using proton magnetic resonance spectroscopy (1H-MRS).Methods 1H-MRS was carried out in 92 healthy Chinese adult volunteers. The metabolite ratios of NAA/Cr, Cho/Cr and NAA/Cho were compared and analysed.ResultsNo significant changes of NAA/Cr, Cho/Cr and NAA/Cho ratios were observed between the left and right precentral gyrus, nor between the male and female. However, the precentral gyrus NAA/Cr and NAA/Cho ratios were decreased gradually with aging and Cho/Cr ratios were increased gradually with aging.ConclusionThe characteristics of the precentral gyrus metabolite concentrations in healthy Chinese adults have been obtained.

Objective To explore the significance of proton magnetic resonance spectroscopy(1H- MRS)in patients with amyotrophic lateral sclerosis(ALS).Methods Single-voxel1 H-MRS was carried on the preeentral gyri in 110 patients with ALS and 24 patients with lower motor neuron syndrome (LMNS) compared with 89 healthy controls.The upper motor neuron involvement of the patients was assessed by upper motor neuron(UMN)signs and the reflex scale,and the disease severity was evaluated by the AIJS function rating scale(ALS-FRS)and APPEL ALS rate scale(AARS).Results Compared with the healthy controls(1.62±0.18),the NAA/Cr of patients with ALS(1.40±0.25)remarkably decreased (t= -5.007,P=0.000),however,it did not change in patients with LMNS(1.60±0.17)as compared with that of the controls.The NAA/Cr of patients with ALS was also lower than that of patients with LMNS(t= -2.660,P=0.009).Furthermore,the NAA/Cr of patients with definite ALS was lower than those of patients with probable and possible ALS(definite vs probable:t=-2.626,P=0.010;definite vs possible: t=-2.537,P=0.013).On the other hand,patients with pronounced UMN signs had a lower NAA/Cr ratio than those with probable UMN signs(t=-2.827,P=0.006).In patients with asymmetric clinical presentation,the NAA/Cr ratio was significantly lower in the precentral gyrus contralateral to the clinically predominantly affected side(t=-2.652,P=0.011).The NAA/Cr was correlated with the reflex scale scores,ALS-FRS score,and AARS and its sub-items(P＜0.05). Conclusions 1H-MRS is useful in studying ALS.The marker NAA/Cr of precentral gyrus may reflect the UMN degeneration in ALS,which is related to the disease severity and progression.However,it might not be helpful in the early diagnosis and differential diagnosis of the disease.

Diabetes is a prevalent disease among the elderly population.Among the antidiabetic drugs, Metformin is the most commonly used, and has been found to have unique anti-aging effects through nutritional sensing regulation, maintenance of protein homeostasis, protection of mitochondrial function, alteration of intercellular communication, and telomere protection, among other mechanisms.The results of studies have shown that Metformin has significant anti-aging potential, thereby necessitating a thorough analysis and summary of its molecular mechanism and research progress in this regard.

Sodium-glucose cotransporter 2 inhibitors(SGLT2i)offer numerous benefits for patients with type 2 diabetes.These benefits include lowering blood pressure and glucose levels, regulating blood lipids, improving arteriosclerosis and endothelial function, reducing urinary protein and decreasing uric acid levels.Additionally, SGLT2i has shown positive outcomes in reducing the risk of kidney failure and cardiovascular adverse events in non-diabetic patients.Recent studies have also discovered that SGLT2i has anti-aging effects, which can potentially delay aging and improve age-related diseases.This article provides a comprehensive review of the potential mechanisms of SGLT2i in anti-aging, offering valuable insights for anti-aging strategies and the prevention of aging-related diseases.

Objective To screen the influencing factors of hepatitis C cirrhosis patients progressing to hepatocellular carcinoma(HCC)using commonly used laboratory testing indicators,establish a prediction model using these indicators and validate them.Methods A total of 231 patients with hepatitis C cirrhosis and 179 patients with hepatitis C HCC hospitalized at the First Affiliated Hospital of Xi'an Jiaotong University between June 2020 and May 2023 were enrolled as the training set,and 105 patients with hepatitis C cirrhosis and 86 patients with hepatitis C HCC hospitalized between June 2023 and February 2024 were enrolled as the validation set.The routine laboratory test indexes of the study subjects in the two groups within the training set were compared,and logistic regression analysis was applied to screen the independent predictors of hepatocellular carcinoma occurrence.Receiver operating characteristic(ROC)curve was used to construct the curve model and validate the model.Results The age,male ratio,ALT,AST,AFP,WBC,NEU,MO,PLT,MPV,PDW,Fbg,NLR and PLR levels of the HCC group were higher than those of the cirrhosis group in the training set(H=-9.07～-2.19),while the levels of INR and LMR were lower than those of the cirrhosis group(H=-4.49,-2.65),and the differences were significant(all P＜0.05).The differences in TP,eGFR,LY and AST/ALT values between the two groups of patients were not significant(H=-1.46～-0.15,all P＞0.05).Multifactorial Logistic regression analysis showed that age(OR=1.048,95％CI:1.023～1.074),Male(OR=1.467,95％CI:1.413～1.765),AST(OR=1.010,95％CI:1.002～1.019),NEU(OR=1.186,95％CI:1.018～1.382)and Fbg(OR=2.245,95％CI:1.639～3.076)were independent risk factors for hepatocellular carcinoma patients(all P＜0.05),and these five independent risk factors were used to construct the HCC column-line graph prediction model,with the AUC for the training set and the validation set AUC(95％Cl)were 0.813(0.771～0.854)and 0.712(0.639～0.784),respectively,and the Hosmer-Lemeshow test showed a good fit of the model with P=0.650 for the training set and P=0.310 for the validation set.Conclusion The prediction model of HCC based on age,gender,AST,NEU and Fbg can have good predictive efficacy and clinical application value.

Objective To explore the correlation between the expression of long non-coding RNA(lncRNA)brain ischemia-related factor(BIRF)and focally amplified lncRNA on chromosome 1(lncRNA FAL1)in serum and the degree of white matter lesions(WML)in patients with cerebral small vessel disease(CSVD).Methods From June 2021 to June 2023,102 CSVD patients admitted to Affiliated Hospital of Chengde Medical University were collected,and these patients were grouped into WML group(n=72)and non WML group(n=30)based on WML diagnostic criteria.According to the Fazekas score,the WML group was further grouped into mild WML group(n=24),moderate WML group(n=36)and severe WML group(n=12).Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was applied to detect the levels of lncRNA BIRF and lncRNA FAL1 in serum.Pearson correlation was applied to analyze the correlation between serum lncRNA BIRF and lncRNA FAL1 levels.Receiver operating characteristic(ROC)curve was applied to analyze the diagnostic value of serum lncRNA BIRF and lncRNA FAL1 levels for severe WML in CSVD patients.Results The age(70.50±5.86 years),history of hypertension(Yes/No,43/29),history of diabetes(Yes/No,45/27),IL-33(68.35±6.80 pg/ml),IL-18(97.78±9.65 ng/L),ubiquitin carboxyl terminal hydrolase L1(UCH-L1)(0.29±0.10 μg/L)and lncRNA BIRF level(2.45±0.30)of patients in the WML group were higher than those in the non WML group(67.10±5.76 years,11/19,9/21,62.48±6.13 pg/ml,92.56±9.37 ng/L,0.24±0.06 μg/L,1.02±0.11),while the expression of serum lncRNA FAL1(0.52±0.10)was lower than that in the non WML group(1.04±0.15),with significant differences(t=2.683,4.518,8.978,4.085,2.510,2.550,25.346,20.500,all P<0.05).The serum lncRNA BIRF levels of CSVD patients in the mild,moderate and severe WML groups(2.23±0.23,2.47±0.31,2.82±0.42)were increased sequentially,while the expression of serum lncRNA FAL1(0.60±0.15,0.51±0.09,0.40±0.04)was decreased sequentially,with significant differences(F=14.913,13.899,all P<0.05).Pearson correlation analysis,the serum levels of lncRNA BIRF and lncRNA FAL1 in patients with WML were negatively correlated(r=-0.603,P<0.001),serum lncRNA BIRF was positively correlated with Fazekas score in WML patients(r=0.483,P<0.001),but serum lncRNA FAL1 was negatively correlated with Fazekas score(r=-0.507,P<0.001).The AUCs of serum lncRNA BIRF and lncRNA FAL1 levels alone and both combination for predicting severe WML in CSVD patients were 0.756(0.641～0.850),0.839(0.733～0.915)and 0.892(0.796～0.953),respectively,and the combination of the two was superior to the detection of serum lncRNA BIRF alone(Z=2.111,P=0.035).Conclusion The serum lncRNA BIRF level is increased and lncRNA FAL1 is reduced in patients with CSVD and WML,and both are related to the degree of WML in CSVD patients.

Objective To compare the short-term clinical effects of totally thoracoscopic surgery and traditional open-heart sugery in the treatment of esophageal cancer. Methods 64 patients with middle thoracic esophageal cancer admitted in our hospital from June 2013 to June 2016 were selected. Among them, 32 patients underwent totally thoracoscopic surgery were identified as totally thoracoscopic group and the remaining 32 patients underwent traditional open-heart surgery were identified as control group. We compared various surgical indicators of patients in the two groups. Results In the totally thoracoscopic group, the duration of operation was(123. 8±25. 1) min, the amount of blood loss during surgery was (172. 1±30. 3) mL, the retention time of chest drainage tube was (3. 1±1. 1) d and the duration of hospitalization was (15. 6±2. 7) d. Compared with the control group, these indicators showed significant difference(P＜0. 05). The incidence of postoperative complications in the totally thoracoscopic group was 9. 3％, which was significantly lower than that in the control group(31. 3％)(P＜0. 05). The pain severity of totally thoracoscopic group reduced significantly than the control group(P＜0. 05). Conclusion Compared with the traditional open-heart surgery, totally thoracoscopic surgery for esophageal cancer has advantages of less bleeding, less trauma, few complications and less pain, which is worthy of promoting and using widely in clinic.