AIM:To construct eukaryotic expression vector of Sirt2 and detect its expression in HEK293 cells. METHODS: Total RNA was isolated from brain tissue of a-dult SD rat. A 1 130 bp fragment containing the coding region of Sirt2 was amplified by RT-PCR and the resulting PCR product was subcloned into PMD20-T vector and se-quenced. Coding region of Sirt2 was generated with PCR by using the PMD20-T-Sirt2 as template, the amplified PCR fragment was inserted into the EcoR I and Hind Ⅲ sites of the pcDNA3. 1myc-his(-)A expression vector, and the sequence was confirmed by DNA sequencing. The expression of new construct pcDNA3.1 myc-his(-) A-Sirt2 in HEK293 cells was detected by immunofluorescence. RESULTS: The full length coding region of Sirt2 was obtained and confirmed by sequencing, the expression of Sirt2 was detected successfully in HEK293 cells. CONCLUSION: The eukaryotic expression vector of Sirt2 has been successfully constructed, which will provide a useful tool for designing an in-depth investigation of the role of Sirt2.

Objective To investigate SNP and distribution of haplotypes in differentially m ethylated region (DMR) upstream of H19 gene in Chinese Korean nationality in order to provide basic data for forensic application and population genetics research. Methods One hundred and one blood sam ples from unrelat-ed Chinese Korean individuals and 14 blood sam ples from 5 Chinese Korean intergenerational fam ilies which known genetic relationship were collected. The SNP in DMRupstream of H19 gene were investi-gated by PCR-cycle sequencing and McrB Cdigestion followed by PCR . The haplotypes detected by parentally im printed allele (PIA) m ethod and relevant genetic param eters were calculated. Results Thirteen SNPs (rs10840167, rs2525883, rs12417375, rs4930101, rs2525882, rs2735970, rs2735971, rs11042170, rs2735972, rs10732516, rs2071094, rs2107425, and rs4930098) and five haplotypes were detected in 1 174 bp target product in DMRupstream of H19 gene, with 9 SNPs having high discrim ination power as good genetic markers. The average gene diversity (GD) of haplotypes was 0.714. The maternal haplotype was confirm ed correctly by PIAm ethod from McrB C-digested products of genom ic DNA. Conclusion High polym orphism sexist in DMRupstream of H19 gene in Chinese Korean nationality. And determ ination of the maternal haplotype could furtherm ore enhance the forensic identification efficiency of im printed gene.

AIM:To study the role of amifostine on the formation of benzo [a]pyrene (BaP)-induced abdomi-nal aortic aneurysm ( AAA) in C57BL/6J mice and the underlying mechanism .METHODS: RAW246.7 mononuclear macrophage in vitro were divided into control group , DMSO group, BaP group, low dose (1 μmol/L) amfostine treated group, middle dose (5 μmol/L) amfostine treated group and high dose (25μmol/L) amfostine treated group .The influ-ence of BaP on the expression of matrix metalloproteinase (MMP)-9, MMP-12, TNF-α, NF-κB in the RAW246.7 mono-nuclear macrophages in vitro was determined by Western blot .Male C57BL/6J mice (8 months old) were divided into con-trol group, model group (AngII+BaP group), low dose (50 mg/kg) amfostine treated group and high dose (100 mg/kg) amfostine treated group.After 6 weeks, the abdominal aorta were isolated .The aortic tissues were subjected to HE and Masson staining.The vascular wall structure , infiltration of macrophage , the expression of MMP-9, MMP-12, TNF-α, NF-κB were evaluated by Western blot and immunochemistry staining .RESULTS:Amifostine attenuated BaP-induced expres-sion of TNF-α, MMP-9, MMP-12, NF-κB in the RAW246.7 mononuclear macrophages (P<0.05).The results of animal experiments showed that the incidence of AAA in high dose amifostine treated group were significantly lower than that in low dose amifostine treated group and model group (P<0.05).Immunohistochemistry staining observation showed that amifos-tine inhibited the aortic macrophage infiltration more obviously in high amifostine treated group compared with model group and low dose amifostine treated group (P<0.05).Compared with model group and low dose amifostine treated group , the MMP-9, MMP-12, TNF-αand NF-κB expression of abdominal aorta in high amifostine treated group was reduced signifi -cantly ( P<0.05 ) .CONCLUSION: Amifostine inhibits BaP-induced activation of macrophages , and also prevents the formation of abdominal aortic aneurysm in C 57BL/6J mice induced by BaP by inhibition of the NF-κB pathway, macro-phage infiltration and the expression of TNF-αand MMPs.

Objective: To investigate the efficacy and safety of daclatavir combined with sofosbuvir treatment in chronic hepatitis C (CHC) in the real world. Methods: A total of 56 CHC patients administrated with daclatavir (60 mg/d) combined with sofosbuvir (400 mg/d) in Huzhou Central Hospital from February to June in 2018 were enrolled. All patients were administrated with daclatavir combined with sofosbuvir for 12 weeks and followed up for 24 weeks. The virological response and the effect of antiviral therapy on hepatic fibrosis were analyzed. Non-structural protein 5A (NS5A) region mutation sequence was detected by Sanger method. Safety and the adverse events were observed. The t test, chi-square test and Mann-Whitney U test were used to analyze the data. Results: Hepatitis C virus (HCV) RNA of all patients treated with daclatavir and sofosbuvir was undectable after eight-week treatment. Sustained virological response at 12 weeks post-treatment (SVR12) was 98.1% (52/53). Gender, globulin, insulin, triglyceride and hemoglobin were correlated with virus clearance (χ²= 4.47, t=2.51, U=1.98, U=2.32 and t=2.03, respectively, all P<0.05). At 12 weeks of the end of the treatment, serum procollagen type Ⅲ, collagen type Ⅳ, liver stiffness measurement (LSM) value, aspartate aminotransferase-to-platelet ratio Index (APRI) score and fibrosis index based on the four factors (FIB-4) score were all improved (U=2.03, 2.15, 2.19, 2.12 and 2.26, respectively, all P<0.05). At 24 weeks of the end of treatment, these indexes showed clear improvement (U=2.09, 2.28, 2.24, 2.33 and 2.46, respectively, all P<0.05). Eight patients had Y93H or L31M resistance-related substitution mutations. Two out of eight patients with variants had negative coversion of HCV RNA and 44.4% (20/45) showed no variants after two weeks of treatment. There was no significant difference (χ² = 1.11, P > 0.05). During the treatment, patients developed dizziness, fatigue, nausea and vomiting, panic, insomnia, sleepiness and sexual function enhancement. Conclusion Daclatavir in combination with sophobuvir shows high virological response and good safety in the treatment of chronic hepatitis C, and liver fibrosis is improved after clearance of HCV.

Objective To explore the operative technique and clinical results of posterior tibial artery perforator flap within saphenous nerve branch for sensory reconstruction.Methods From January,2016 to June,2018,9 patients suffered from soft tissue defect were treated by the posterior tibial artery perforator flap containing saphenous nerve branch.Seven patients were males and 2 were females,with age ranged from 31 to 62 years.Soft tissue defects located in hands in 5 patients,plantar in 2 patients,ankle in 1 patient and dorsal foot in 1 patient.The size of soft tissue defects ranged from 8.0 cm×2.5 cm to 21.0 cm×4.0 cm.The regular post-operative followed-up was performed.Results All flaps survived without complications.The size of flap ranged from 10.0 cm×3.5 cm-23.0 cm×5.0 cm.Donor sites were primarily closed in 5 patients and secondary closed in 4 patients.Followed-up ranged from 6 to 15 months with 10 months in average.The contour of flaps were satisfied and the sensory function of the donor sites were normal.At 6 months followed-up,SW test reached 5.07 in all flaps,and 2PD ranged from 14 to 35 mm.Conclusion The novel sensory flap can provide satisfied sensory outcome without sacrificing main artery and saphenous nerve,and is a good candidate for sensory reconstruction of soft tissue defects.

Objective To evaluate the effect of latanoprost on cell proliferation of and melanogenesis in human epidermal melanocytes,and to explore its mechanism.Methods Latanoprost was added into the 254 medium to prepare latanoprost solutions at different concentrations of 10-5,10-6 and 10-7 mol/L.In vitro cultured human epidermal melanocytes were divided into 4 groups to be cultured with media containing no latanoprost (control group) or 10-5,10-6 and 10-7 mol/L latanoprost for 48 hours.Cell counting kit-8 (CCK8) assay was performed to evaluate the proliferative activity of melanocytes,dopa oxidation assay to estimate the activity of tyrosinase.Sodium hydroxide (NaOH)-lysis method was used to determine the content of melanin,and Masson-Fontana staining to observe the number and distribution of melanin granules.Westernblot analysis and real-time fluorescence-based quantitative PCR were performed to determine the protein and mRNA expression of melanogenesis-related genes including microphthalmia-associated transcription factor (MITF),tyrosinase (TYR) and tyrosinase-related protein 1 (TYRP1).Comparison among the 4 groups and multiple comparisons were done by using one-way analysis of variance and least significant difference (LSD)-t test.Results Compared with the control group,the 10-6-,10-5-mol/L latanoprost groups showed significantly increased proliferative activity of melanocytes (1.064 ± 0.172 and 1.078 ± 0.080 vs.0.784 ± 0.015;t =3.289,3.454 respectively,both P ＜ 0.05),increased activity of tyrosinase (0.510 ± 0.017 and 0.454 ± 0.009 vs.0.355 ± 0.041;t =6.139,3.939 respectively,P ＜ 0.01 or 0.05),and increased content of melanin (t =7.232,5.967,both P ＜ 0.01).However,there were no significant differences in the proliferative activity of melanocytes,activity of tyrosinase or content of melanin between the 10-7-mol/L latanoprost group and control group (all P ＞ 0.05).Masson-Fontana staining showed more and darker melanin granules on melanocyte dendrites in the 10-5-,10-6-,10-7-mol/L latanoprost groups than in the control group,and the color of melanin granules changed from light brown to black brown along with the increase in the concentration of latanoprost.The mRNA expression of MITF increased along with the increase in the concentration of latanoprost (P ＜ 0.01),and the protein expression of MITF wassignificantly higher in the 10-6,10-5-mol/L latanoprost groups than in the control group and 10-7-mol/L latanoprost group (all P ＜ 0.01).The 10-6-mol/L latanoprost group showed significantly increased mRNA and protein expression of TYR and TYRP1 compared with the control group,10-7-,10-5-mol/L latanoprost groups (all P ＜ 0.01).Conclusion Latanoprost can increase the proliferation of human epidermal melanocytes,and promote tyrosinase activity and melanogenesis likely by enhancing the mRNA and protein expression of MITF,TYR,TYRP1.

OBJECTIVETo determine the incidence and risk factors for in-hospital stroke in patients with acute myocardial infarction (AMI).

METHODSTwo thousand one hundred and thirty-three patients with acute myocardial infarction were studied retrospectively. History, demographic, clinical, thrombolytic and anticoagulant data of AMI were obtained through review of the medical record. The relative risk of in-hospital stroke in patients with AMI was estimated using multiple nonconditional logistic regression to adjust for potential confounding factors.

RESULTSAmong 2,133 consecutive patients admitted with acute myocardial infarction to hospitals in Shandong and Hubei provinces, 98 (4.59%) experienced strokes during hospitalization. In our analysis, anterior infarction was the most important risk factor for in-hospital stroke (ischemic and hemorrhagic) in patients with AMI (RR = 7.04), followed by history of hypertension (RR = 3.41), previous stroke (RR = 1.69), atrial fibrillation (RR = 2.22), advanced age (RR = 1.04), and higher heart rate at enrollment (RR = 1.03).

CONCLUSIONSThe incidence of in-hospital stroke in patients with AMI increased with lower rate of using thrombolytic agents. Thrombolytic therapy was noticed as a protective factor for stroke after AMI. The risk factors for in-hospital stroke with AMI were history of hypertension, previous stroke, atrial fibrillation, advanced age, anterior infarction, and higher heart rate at enrollment.

Adult ; Aged ; Female ; Humans ; Incidence ; Male ; Middle Aged ; Multivariate Analysis ; Myocardial Infarction ; complications ; Risk Factors ; Stroke ; epidemiology ; etiology

Objective:To investigate the clinical efficacy of laparoscopic hepatic caudate lobectomy.Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 5 patients with hepatic caudate lobe tumor who were admitted to the General Hospital of PLA Northern Theater Command from July 2018 to June 2021 were collected. There were 2 males and 3 females, aged 49(range, 26-55)years. All 5 patients underwent laparoscopic hepatic caudate lobectomy. Observation indicators: (1) intraoperative situations; (2) postoperative situations; (3) follow-up. Follow-up was conducted using telephone interview or outpatient examination to detect tumor recurrence. The follow-up was up to March 2023. Measurement data with skewed distribution were represented as M(range) and count data were described as absolute numbers. Results:(1) Intraoperative situations. All 5 patients underwent laparoscopic hepatic caudate lobectomy with the left approach, including 3 cases receiving laparoscopic partial resection of hepatic caudate lobe, 1 case receiving laparoscopic partial resection of hepatic caudate lobe+left lateral lobectomy and 1 case receiving laparoscopic partial resection of hepatic caudate lobe+cholecystectomy. Intraoperative hilar occlusion was performed in 3 patients. The operation time and volume of intraoperative blood loss of 5 patients was 240(range, 180-370)minutes and150(range, 100-200)mL, respectively. (2) Postoperative situations. On the first day after surgery, the levels of albumin, alanine aminotransferase, and aspartate aminotransferase in 5 patients was 32.9(range, 29.2-40.3)g/L, 104.09(range, 57.11-1 018.67)U/L, and 67.13(range, 58.00-852.66)U/L, respectively. Three of 5 patients showed no significant changes in prothrombin time (PT) on the first day after surgery, while two patients did not undergo PT test. On the third day after surgery, the visual analogue score of 5 patients was mild pain. Among the 5 patients, 1 case had anal exhaust on the postoperative 1st day, and 4 cases had anal exhaust on the postoperative 2nd day. The time to drainage tube removal for 5 patients was 6(range, 4-10)days. There was no postoperative complication such as bleeding, infection, bile leakage or liver failure in 5 patients. Results of postoperative histopathological examination showed that 2 cases with adenoma, 2 cases with cavernous hemangioma and 1 case with smooth myolipoma. The duration of postoperative hospital stay of 5 patients was 6(range, 5-11)days. (3) Follow-up. All 5 patients were followed up for 22 (range, 19-51)months, and there was no tumor recurrence.Conclusion:It is safe and feasible to selectively carry out laparoscopic hepatic caudate lobectomy by strictly gras-ping the indications.

Objective:To compare the efficacy of lower extremity three dimensional CT venography (CTV) and lower extremity ascending phlebography in evaluating recurrent varicose veins.Methods:A retrospective analysis was conducted on clinical data from 235 patients with unilateral recurrent varicose veins who were treated at the Department of Vascular Surgery,Beijing Shijitan Hospital,Capital Medical University, between January 2015 and December 2020.There were 112 males and 123 females, with an age of (62.5±11.4)years (range:24 to 75 years).Patients were stratified into two groups based on preoperative imaging examination:the CTV group (utilizing lower extremity venous ultrasound+lower extremity CTV) and the control group (employing lower extremity venous ultrasound+lower extremity ascending phlebography).The two groups were matched in a 1∶1 ratio using propensity score matching, resulting in 43 cases per group.Comparative analyses between the groups at the one-year postoperative follow-up were performed using independent sample t tests, Wilcoxon rank-sum tests, χ2 tests, and linear regression analysis. Results:One year post-surgery,the CTV group exhibited a lower venous clinical severity score (VCSS) compared to the control group( M(IQR),3.0(4.3) vs.4.0(5.8), Z=-2.038, P=0.040).Additionally, the chronic venous insufficiency patients′ quality of life questionnaire (CIVIQ-20) scores were significantly higher in the CTV group than in the control group (89.0(8.0) vs.82.5(17.0), Z=-2.627, P=0.010).Patients in the CTV group also experienced a shorter ulcer healing time compared to the control group (4.0(4.0) weeks vs.12.0(7.0) weeks, Z=-3.217, P<0.01).Both groups showed no clinically symptomatic recurrent varicose veins or ulcers.However, they exhibited ultrasound-detectable varicose vein recurrence, with no statistically significant difference ( χ2=0.453, P=0.500).The number of diseased vessels requiring management based on ultrasound supplemented by CTV was 16, while the number supplemented by ascending phlebography was 7,with a statistically significant difference ( χ2=4.800, P=0.030).Linear regression analysis demonstrated that clinical-etiology-anatomy-pathology clinical grading and the preoperative imaging examination method exerted independent influences on VCSS and CIVIQ-20 during the one-year postoperative assessment. Conclusions:CTV-assisted ultrasound enables a direct and comprehensive evaluation and localization of diseased veins in patients with recurrent varicose veins.The utilization of lower extremity vein ultrasound combined with CTV-guided management of lower extremity vessels in minimally invasive treatment significantly improves patient prognosis, surpassing the assessment provided by ascending phlebography.

Hepatocellular carcinoma is one of the most common malignant tumors in the world, which is a serious threat to human health. HBV infection is one of the most common causes of hepatocellular carcinoma.The diagnosis of most hepatocellular carcinoma has progressed to the middle and late stage, and the prognosis is poor. Early detection, diagnosis and treatment are important supports to improve the clinical outcome of hepatocellular carcinoma. In recent years, scholars at home and abroad have established various hepatocellular carcinoma risk prediction models, which are conducive to improving the early diagnosis rate of hepatocellular carcinoma and reducing the mortality rate. This article reviews the risk factors and risk prediction models of chronic hepatitis B associated hepatocellular carcinoma, in order to provide reference for HBV-associated liver cancer risk monitoring and management decision.