Objective To construct Cox7a2 fluorescent vector and study its effect on cytochrome C oxidase (COX) activity in mouse Sertoli cell line TM4. Methods The coding region of Cox7a2 was amplified from mouse Sertoli cell line TM4 by RT-PCR. The PCR product was inserted into pEYFP-C1 vector with BamH I and EcoR I restriction site, and confirmed by DNA sequencing. The recombinant fusion protein vector was amplified by transforming into DH5a and transfected into TM4 cells. The protein expression was identified by Western blot. COX activity was measured by spectrophotometer 6, 12, 24 and 48 h after the transfection of recombinant vector into the TM4 cell line. Results The entire coding sequence of Cox7a2 was cloned with 252 bp length. Plasmid pEYFP-C1-Cox7a2 vector was constructed and the positive clones were verified by restriction enzymes digestion and DNA sequencing. The transfection efficiency of the TM4 cell line was about 70% and 37000 D fusion protein was obtained. The COX activities were (0.642±0.051), (0.542±0.049), (0.311±0.021) and (0.216±0.010) U/mg 6, 12, 24 and 48 h after the transfection of recombinant vector in the TM4 cell line. Meanwhile, the COX activities were (0.714±0.064) and (0.653±0.031) U/mg in non-tranfected and naked vector group respectively. Compared with the non-tranfected group, COX activity decreased significantly 12, 24 and 48 h after the transfection. Conclusions The recombint plasmid vector was successfully constructed. Cox7a2 gene has an inhibiting effect on COX activity and may play an important role in the regulation of COX activity in mouse Sertoli cell line TM4.

Dehydroascorbate reductase (DHAR) plays an important role in the recycling of ascorbic acid. In this work, we isolated the full length cDNA clones of two different DHAR genes (tentatively named as TaDHAR1 and TaDHAR2, respectively) from common wheat. Semi-quantitative PCR experiments showed that TaDHAR1 and TaDHAR2 were transcribed in many vegetative and reproductive organs examined in this work. Transient expression analysis using wheat protoplasts indicated that the protein products of TaDHAR1 and TaDHAR2 may be located in the cytoplasm. The cDNAs of TaDHAR1 and TaDHAR2 were expressed in the bacterial cells, and resultant histidine tagged recombinant proteins could be efficiently purified using nickel chelate affinity chromatography. In vitro enzyme activity assays revealed that the recombinant TaDHAR1 and TaDHAR2 proteins could all convert dehydroascorbate (DHA) to AsA. The two proteins exhibited higher activity levels at 37 degrees C than at 25 degrees C. Under the two temperature conditions, the optimal pH for TaDHAR1 and TaDHAR2 was both around 7.5. The major difference between TaDHAR1 and TaDHAR2 is the activity under pH 6.0 and 7.0 at 25 degrees C. The results and resources obtained in this study may be useful for further research into the physiological role of TaDHAR genes in AsA metabolism in crop plants under normal or stressed conditions.
Amino Acid Sequence ; Cloning, Molecular ; Genes, Plant ; genetics ; Molecular Sequence Data ; Oxidoreductases ; genetics ; metabolism ; Plant Proteins ; genetics ; metabolism ; Recombinant Proteins ; genetics ; metabolism ; Triticum ; enzymology ; genetics

OBJECTIVE: To understand the quality of life and related factors among family members of tuberculosis patients and provide a reference for the improvement of their quality of life. METHODS: A total of 222 family members of tuberculosis patients at 4 tuberculosis hospitals in Changsha and 327 healthy controls were surveyed with structured questionnaire, the short version of the WHO quality of life scale (WHOQOL-BREF). RESULTS: The mean score of the family members of tuberculosis patients in the psychological domain, physical domain and environmental domains was lower than that of the control group (P<0.01). Multiple linear regression showed that gender, age, monthly income, educational level, patient condition and knowledge of tuberculosis prevention and treatment were the factors affecting their quality of life. CONCLUSION The quality of life of the family members of tuberculosis patients is lower than that of the control group.
Family Health ; Humans ; Quality of Life ; Surveys and Questionnaires ; Tuberculosis ; psychology

Objective To evaluate the effectiveness and safety of flexible ureteroscopy lithotripsy (FURL) combined with holmium laser for the concurrent treatment of bilateral upper ureteral tract calculi.Methods The clinical data in 43 patients with bilateral upper ureteral tract calculi concurrently treated by FURL combined with holmium laser in our hospital from September 2014 to November 2016 were retrospectively analyzed.All cases conducted CT scanning before operation.The flexible ureteroscope was intraoperatively placed and the lithotripsy was conducted by using 200 μm optical fiber at a power of 0.8-1.0 J/10-20Hz,which was coordinated by reticular basket.After lithotripsy,bilateral urethral stents were placed.KUB or CT was re-examined on postoperative 1 d.The cases of residual stone were performed KUB again after removing urethral stent after 4-6 weeks.Results All cases were successfully placed the flexible ureteroscope by one time.The mean operation time was (101.5±37.2) min and the overall stone-free rate (SFR) was 81.4 ％ (35/43).SFR was 100％ in the patients with the stone burden less than 30 mm and 63.2％ in the patients with the stone burden ≥30 mm,the difference was statistically significant(P＜0.05).One case appeared fever (39.5 ℃) after operation,1 case developed subcapsular renal hematoma and no other severe complications occurred.Conclusion FURL combined with holmium laser is an effective means to concurrently treating bilateral upper ureteral calculi,especially for the patients with stone burden ＜30 mm,which has higher stone clearance rate with good safety.

Objective:To investigate the clinical value of esophageal-jejunal OrVil TM anas-tomosis and Overlap anastomosis in laparoscopic radical total gastrectomy of adenocarcinoma of esophagogastric junction (AEG). Methods:The retrospective cohort study was conducted. The clinicopathological data of 112 patients with AEG who were admitted to the First Hospital of Jilin University from July 2017 to August 2022 were collected. There were 87 males and 25 females, aged (64±8)years. All 112 patients underwent laparoscopic total gastrectomy and D 2 lymphadenectomy, in which 61 cases with esophageal-jejunal OrVil TM anastomosis were divided into the OrVil TM group, 51 cases with esophageal-jejunal Overlap anastomosis were divided into the Overlap group. Observa-tion indicators: (1) surgical situations; (2) postoperative complications; (3) influencing factors for patients undergoing esophageal-jejunal OrVil TM anastomosis. Measurement data with normal distri-bution were represented as Mean± SD, and comparison between groups was conducted using the independent sample t test. Measurement data with skewed distribution were represented as M( Q1, Q3), and comparison between groups was conducted using the non-parameter test. Count data were described as absolute numbers, and comparison between groups was conducted using the chi-square test or Fisher exact probability. Logistic regression model was used for multivariate analysis. Results:(1) Surgical situations. The esophageal invasion length and tumor diameter was 1.0(0.7,2.0)cm and (6.3±2.7)cm in patients of the OrVil TM group, versus 0.2(0.1,0.5)cm and (4.7±2.2)cm, respectively, in patients of the Overlap group, showing significant differences in the above indicators between the two groups ( Z=?6.14, t=3.26, P<0.05). (2) Postoperative complications. Cases with complications ≥Ⅲa grade of Clavien-Dindo classification, cases with respiratory system complications, cases with hydrothorax were 13, 17, 13 in the OrVil TM group, versus 4, 5, 4 in the Overlap group, showing significant differences in the above indicators between the two groups ( χ2=3.91, 5.74, 3.91, P<0.05). Cases underwent readmission within postoperative 30 days were 3 and 1 in the OrVil TM group and the Overlap group, respectively, and all patients recovered after symptomatic treatment. There were 2 cases died after operation in the OrVil TM group and none of patients died after operation in the Overlap group. (3) Influencing factors for patients undergoing esophageal-jejunal OrVil TM anastomosis. Results of multivariate analysis showed that esophageal invasion length was an independent factor influencing for patients undergoing esophageal-jejunal OrVil TM anastomosis ( odds ratio=8.25, 95% confidence interval as 3.41?19.96, P<0.05). Conclusions:Compared with esophageal-jejunal Overlap anastomosis, choosing the esophageal-jejunal Orvil TM anastomosis during laparoscopic radical total gastrectomy can take benefit to the proximal margin of patients with AEG. However, the ratios of complications ≥ Ⅲa grade of Clavien-Dindo classification, respiratory system complications and hydrothorax associated to OrVil TM anastomosis are relatively increased. Esophageal invasion length is an independent influencing factor for patients undergoing esophageal-jejunal OrVil TM anastomosis.

Objective:To investigate associations of cerebral perfusion impairment with early neurological deterioration (END) and poor outcome in patients with acute small subcortical infarction (SSI).Methods:Patients with SSI in the perforator artery region admitted to the Department of Neurology, Zhengzhou People's Hospital between January 2020 and November 2022 were prospectively included. END was defined as an increase of ≥2 in the total score of the National Institutes of Health Stroke Scale (NIHSS) or an increase of ≥1 in the motor function score within 72 h after admission. Poor outcome was defined as the modified Rankin Scale score of 2 at 90 d after onset. Cerebral perfusion impairment was defined according to MRI perfusion-weighted imaging parameters. The demographic, baseline clinical and imaging data were collected. Multivariate logistic regression analysis was used to determine associations of cerebral perfusion impairment and END and poor outcome in patients with SSI. Results:A total of 100 patients with SSI were enrolled, including 56 males (56.0%), and aged 69.2±5.8 years. Among them, 19 patients (19.0%) developed END, 27 (27.0%) had poor outcome, and 51 (51.0%) had significant cerebral perfusion impairment. There were statistically significant differences in high sensitivity C-reactive protein, white matter hyperintensities (WMHs) in the basal ganglia, enlarged perivascular space (EPVS) in the basal ganglia, deep cerebral microbleeds (CMBs), and cerebral perfusion impairment between the END group and the non-END group (all P<0.05). Multivariate logistic regression analysis showed that higher diastolic blood pressure (odds ratio [ OR] 1.070, 95% confidence interval [ CI] 1.003-1.141); P=0.040], deep WMHs ( OR 2.271, 95% CI 1.135-4.544; P=0.020), deep CMBs ( OR 5.047, 95% CI 1.240-20.549; P=0.024), and cerebral perfusion impairment ( OR 6.083, 95% CI 1.318-28.080; P=0.021) were independent risk factors for END in patients with SSI. There were statistically significant differences in hypersensitive C-reactive protein, NIHSS score at END, basal ganglia EPVS, END, and cerebral perfusion impairment between the poor outcome group and the good outcome group ( P<0.05). Multivariate logistic regression analysis showed that NIHSS score at END ( OR 1.485, 95% CI 1.034-2.133; P=0.032), basal ganglia EPVS ( OR 3.005, 95% CI 1.224-7.378; P=0.016), and cerebral perfusion impairment ( OR 9.234, 95% CI 1.994-42.765; P=0.004) were independent risk factors for the poor outcome at 90 d in patients with SSI, while anterior circulation infarction ( OR 0.066, 95% CI 0.013-0.334; P=0.001) was independently negatively correlated with the poor outcomes at 90 d after onset. Conclusion:Cerebral perfusion impairment is an independent risk factor for END and poor outcome at 90 d after onset in patients with SSI.

Empathy is one of the basic qualities that modern clinicians need to possess.Although traditional Chinese medicine has no definition for empathy,it does have practical applications.TCM clinical empathy,nurtured by Chinese excellent traditional culture,can be seen in TCM classics and clinical practice of famous doctors of previous generations,and has been inherited in the spiritual thinking of modern colleges and universities of Chinese medicine.However,at present,there is a relative lack of special training courses for"empathy"in Chinese medicine higher education.This paper first briefly describes the connotation,history,and neuroscience mechanism of the concept of"empathy".Secondly,the manifestations and characteristics of"empathy"in the clinical practice of traditional Chinese medicine were sorted out.Finally,we use modern medicine and psychology for reference,and combine the characteristics of traditional Chinese medicine to make suggestions for the cultivation of the ability of"empathy"in clinical Chinese medicine.

Uridine diphosphate glucuronic acid transferase (UDP-glucuronosyltransferases, UGT) is an important Ⅱmetabolic enzymes in the body. It is invovled in the metabolism of exogenous compounds, and also in endogenous substances such as bile acid metabolism and regulation. Parsing uridine diphosphate glucuronic acid transferase mediated bile acid metabolism and its influence factors can help enhance related disease treatment and prevention. Studies have shown that the interaction between UGT and bile acids is influenced by many endogenous and exogenous factors. This paper will focus on the effects of internal and exogenous factors such as nuclear receptors, genetic factors, xenobiotics and liver-related diseases on the action of UGT enzyme, and discuss the potential mechanism of bile acid balance intervention.

AIM:Bone morphogenetic protein 7(BMP7)reduces the expression of Yes-related protein 1(YAP1)by down-regulating Ajuba level and decreasing extracellular matrix(ECM)deposition.This study aimed to inves-tigate the influence of these factors on modifying the degree of renal fibrosis in rats with diabetic nephropathy.METH-ODS:Eighteen Sprague-Dawley(SD)rats were randomly divided into three groups:the normal control(NC)group,the diabetes mellitus(DM)group,and the DM group treated with BMP7 overexpressing adeno-associated virus(DM+rAAV-BMP7).Each group consisted of six rats.Diabetic kidney disease(DKD)was established in the DM and DM+rAAV-BMP7 groups by injecting 55 mg/kg streptozotocin(STZ)via the tail vein.NRK-52E cells were divided into three groups:the normal glucose(NG)group,the high glucose(HG)group,and the high glucose group treated with recombinant hu-man BMP7(HG+rhBMP7)group.Pathological changes in renal tissues were observed using hematoxylin and eosin(HE)and Sirius red staining.Immunohistochemical staining was performed to examine the expression sites of Ajuba and YAP1 in the renal cortex.Western blot analysis was conducted to determine the expression levels of BMP7,Ajuba,YAP1,colla-gen type Ⅲ(Col-Ⅲ),and fibronectin(FN)in the rat renal cortex and NRK-52E cells.RT-qPCR was used to measure the mRNA levels of Ajuba and YAP1 in the rat renal cortex.RESULTS:Biochemical indices revealed significantly ele-vated levels of blood glucose,serum creatinine,triglycerides,total cholesterol,and 24-hour urinary protein in the DM group compared to the NC group(P<0.05).In the DM+rAAV-BMP7 group,the levels of serum creatinine,24-hour uri-nary protein,triglycerides,and total cholesterol were lower than those in the DM group(P<0.05).Pathological staining demonstrated that the renal interstitium of the DM group exhibited inflammatory cell infiltration,fibrous tissue,collagen fi-ber deposition,disordered renal tubule arrangement,atrophy,and vacuolar degeneration,which were ameliorated in the DM+rAAV-BMP7 group.Immunohistochemistry revealed that Ajuba and YAP1 were mainly expressed in the cytoplasm and nucleus,with high expression in the cytoplasm of the DM group,which was significantly decreased in the DM+rAAV-BMP7 group.Western blot results indicated that the protein levels of FN,Col-Ⅲ,Ajuba,and YAP1 were up-regulated in the DM and the HG groups(P<0.05),but significantly down-regulated in the DM+rAAV-BMP7 group(P<0.05).RT-qP-CR results demonstrated that the mRNA levels of Ajuba and YAP1 were higher in the DM group and significantly lower in the DM+rAAV-BMP7 group(P<0.05).CONCLUSION:The overexpression of BMP7 can ameliorate renal fibrosis in rats with DKD.This effect is likely mediated by the down-regulation of Ajuba,reduction of YAP1 expression,and subse-quent inhibition of ECM deposition.

【Objective】 To investigate the clinical efficacy and safety of laparoscopic radical prostatectomy in elderly prostate cancer patients aged ≥80 years in good health. 【Methods】 A total of 18 octogenarian patients who underwent laparoscopic radical prostatectomy during Aug.2016 and Mar.2020 at our hospital were selected for retrospective analysis. 【Results】 The patients were 80 to 86 years old. The preoperative serum PSA was 5.25-101.00 ng/mL. There were 2 cases in stage cT1N0M0,12 in stagec T2N0M0 and 4 in stage cT3N0M. The Gleason score was ≤6 in 4 cases,=7 in 9 cases,and ≥8 in 5 cases. During the follow-up of 22 to 65 months,the rates of urinary incontinence at 1 month,3 months,6 months,and 1 year were 50.00% (9/18),33.33% (6/18),16.67% (3/18) and 5.56 %(1/18),respectively. The postoperative urinary control recovery time was 1 to 220 days. Positive incision margin developed in 3 cases and biochemical recurrence occurred in 3 cases. The IPSS score at 3, 6 months and 1 year after surgery were lower than these before surgery (P<0.05). The overall health score of the FACD-P scale at 6 months after surgery was higher than that before surgery (P<0.05). 【Conclusion】 For well selected octogenarian prostate cancer patients in good condition,laparoscopic radical prostatectomy is a feasible,safe and effective treatment option,but long-term follow-up and prospective clinical studies with large sample size are still needed to confirm its efficacy.