BACKGROUND:Oxidative injury is considered to be one of the important factors of cerebral ischemia-reperfusion injury.Superoxide dismutase 2(SOD2)is a key mitochondrial antioxidant molecule,and fenofibrate can regulate the expression of SOD2 by activating peroxisome proliferator-activated receptor α. OBJECTIVE:To explore whether the mechanism of fenofibrate in the treatment of cerebral ischemia-reperfusion injury depends on the expression of SOD2. METHODS:The TALENs system was used to construct SOD2 transgenic mice.The transgenic mice were genotyped by PCR and DNA sequencing techniques.The expression of SOD2 protein in transgenic mice was detected by western blot assay.Wild-type and SOD2 transgenic mice were randomly divided into four groups:wild-type control group(n=6),wild-type fenofibrate group(n=6),SOD2 transgenic control group(n=5)and SOD2 transgenic fenofibrate group(n=5).A mouse model of middle cerebral artery occlusion was prepared using the suture-occlusion method.After 90 minutes of ischemia,the thread was removed to reperfuse cerebral blood flow for 30 minutes.A cerebral blood flow monitor was used to monitor local cerebral blood flow.Brain tissue slices were taken for 2,3,5-triphenyltetrazolium chloride staining to analyze the situation of cerebral infarction in each group. RESULTS AND CONCLUSION:After PCR and DNA sequencing analysis,nine SOD2+/+ transgenic mice were successfully constructed.After cerebral ischemia-reperfusion,the wild-type fenofibrate group showed partial recovery of cerebral blood flow and significantly reduced cerebral infarction volume compared with the wild-type control group(P<0.001).There was no significant difference in cerebral blood flow and cerebral infarction volume between the SOD2 transgenic fenofibrate group and the SOD2 transgenic control group.The SOD2 transgenic control was superior to the wild-type control group in terms of improving cerebral blood flow and cerebral infarction(P<0.001).There were also no significant differences in cerebral blood flow and cerebral infarction volume between the wild-type fenofibrate group and the SOD2 transgenic control group and between the wild-type fenofibrate group and the SOD2 transgenic fenofibrate group.To conclude,the expression of SOD2 is one of the mechanisms of fenofibrate in the treatment of cerebral ischemia-reperfusion injury.

Objective:To explore the genetic characteristics of a family with intellectual impairment and developmental delay caused by HERC2 gene mutation. Methods:A total of 10 individuals from a 3-generation family of a child with intellectual disability and developmental delay who was treated at the First Affiliated Hospital of Dali University from May to July 2020 were recruited. Clinical data of probands from the family and the illness status of family members were collected. Whole exome sequencing technology was used to screen for pathogenic genes in probands, meanwhile, Sanger sequencing was conducted to verify the family of suspected pathogenic genes. According to the American College of Medical Genetics and Genomics (ACMG) genetic variation interpretation standards and guidelines, pathogenicity classification of suspected pathogenic gene mutation sites was performed.Results:The patient, male, 12-year old, presented with the clinical characteristics of "developmental delay for 9 years and intellectual disability for 3 months". The patient was characterized by a short stature, spontaneous laughter and avoidance of the surrounding environment. One younger brother has symptoms similar to intellectual impairment with developmental delay, while the remaining family members are normal. The Wechsler Intelligence Test of the child indicates a low level of intelligence. The whole exome sequencing results showed that the proband carried a homozygous mutation at the c.7675A>G site of the HERC2 gene, while Sanger sequencing showed that the younger brother carried the same homozygous mutation at the c.7675A>G site of the HERC2 gene. The other family members carried the same heterozygous mutation at the c.7675A>G site of the HERC2 gene. According to the ACMG guidelines, the mutation at this gene site is clinically unclear. Conclusion:The mutation at the c.7675A>G locus of HERC2 gene is the genetic basis of mental retardation with stunting in this family.

Objective:To explore the genetic characteristics of a family with intellectual impairment and developmental delay caused by HERC2 gene mutation. Methods:A total of 10 individuals from a 3-generation family of a child with intellectual disability and developmental delay who was treated at the First Affiliated Hospital of Dali University from May to July 2020 were recruited. Clinical data of probands from the family and the illness status of family members were collected. Whole exome sequencing technology was used to screen for pathogenic genes in probands, meanwhile, Sanger sequencing was conducted to verify the family of suspected pathogenic genes. According to the American College of Medical Genetics and Genomics (ACMG) genetic variation interpretation standards and guidelines, pathogenicity classification of suspected pathogenic gene mutation sites was performed.Results:The patient, male, 12-year old, presented with the clinical characteristics of "developmental delay for 9 years and intellectual disability for 3 months". The patient was characterized by a short stature, spontaneous laughter and avoidance of the surrounding environment. One younger brother has symptoms similar to intellectual impairment with developmental delay, while the remaining family members are normal. The Wechsler Intelligence Test of the child indicates a low level of intelligence. The whole exome sequencing results showed that the proband carried a homozygous mutation at the c.7675A>G site of the HERC2 gene, while Sanger sequencing showed that the younger brother carried the same homozygous mutation at the c.7675A>G site of the HERC2 gene. The other family members carried the same heterozygous mutation at the c.7675A>G site of the HERC2 gene. According to the ACMG guidelines, the mutation at this gene site is clinically unclear. Conclusion:The mutation at the c.7675A>G locus of HERC2 gene is the genetic basis of mental retardation with stunting in this family.

OBJECTIVE: To explore the genetic basis of a Chinese pedigree affected with chronic kidney disease (CKD). METHODS: A Chinese pedigree comprised of 10 individuals from four generation who had visited the First Affiliated Hospital of Dali University from August 15, 2018 to July 5, 2021 was selected as the study subject. Clinical data of the proband were collected, and a pedigree survey was conducted. The proband was subjected to whole exome sequencing (WES). Candidate variant was verified by Sanger sequencing and bioinformatic analysis. RESULTS: The proband, a 41-year-old female, has been diagnosed with chronic nephritis for more than 4 years. Routine urinary examination showed proteinuria and blood creatinine of 1 130 μmol/L. Renal biopsy has revealed hyperplastic glomerulonephritis, moderate tubulointerstitial disease and renal arteriosclerosis. Her elder sister, younger brother, younger sister and mother were all diagnosed with CKD stage 5. Except for her elder sister, all of them had deceased, whilst no abnormality was found in the remainders. Genetic testing revealed that the proband and four family members had harbored a c.467G>A missense variant of the PAX2 gene. The variant has been associated with focal segmental glomerulosclerosis and classified as likely pathogenic (PS1+PP3+PP4) based on the guidelines from the American College of Medical Genetics and Genomics (ACMG). CONCLUSION The c.167G＞A variant of the PAX2 gene probably underlay the CKD in this Chinese pedigree.
Adult ; Female ; Humans ; Male ; East Asian People ; Genetic Testing ; Mutation ; PAX2 Transcription Factor/genetics* ; Pedigree ; Renal Insufficiency, Chronic/genetics*

【Objective】 To investigate the relationship between preoperative albumin-to-alkaline phosphatase ratio (AAPR) and postoperative recurrence of localized penile cancer (T1-3N0M0). 【Methods】 Clinical data of patients with limited penile cancer admitted to our hospital during Jan.2012 and Jan.2017 were collected to compare the differences in age, body mass index (BMI), AAPR, hypertension, diabetes mellitus, tumor diameter, postoperative pathological grading and staging between the postoperative recurrence group and the non-recurrence group. 【Results】 The differences in AAPR(P=0.001), WHO/ISUP pathological grading(P=0.018), and pathological stage(P=0.012)between the recurrence and non-recurrence groups were statistically significant. Cox regression results showed that AAPR was an independent risk factor for recurrence (P=0.041). Survival curve results showed that patients in the high and low AAPR groups had an inverse relationship with recurrence-free survival (P=0.028). 【Conclusion】 Preoperative AAPR is an independent risk factor for recurrence after surgery for limited penile cancer and is associated with recurrence-free survival. As AAPR increases, the incidence of recurrence decreases and progression-free survival increases.

Objective:To explore independent risk factors and risk stratification for diagnosis of clinically significant prostate cancer (CsPCa) in biopsy-naive patients with nonsuspicious multiparametric magnetic resonance imaging (mpMRI).Methods:The data of 549 patients who underwent initial systematic biopsy (SB) in the First Affiliated Hospital of Soochow University, the Second Affiliated Hospital of Soochow University and Traditional Chinese Medicine Hospital of Kunshan between October 2015 and January 2021 were retrospectively reviewed. Nonsuspicious mpMRI was defined as Prostate Imaging-Reporting and Data System (PI-RADS)≤2. All patients received systematic 12 core prostate biopsy, 278 of them by transperineal and 271 by transrectal biopsies. The median age of the patients was 67 (62, 73) years, the median prostate specific antigen (PSA) was 9.01 (6.15, 13.64) ng/ml, the median prostate volume was 48.41 (35.85, 64.28) ml, and 54 patients were positive in digital rectal examination (DRE). Taking CsPCa as the outcome index, receiver operating characteristic (ROC) analysis was performed on age, tPSA, f/tPSA and PSA density (PSAD) to obtain the optimal cut-off value, and logistics regression was used to explore the independent risk factor of CsPCa in mpMRI negative patients. The optimal cut-off value when the negative predictive value (NPV) of mpMRI diagnosis of CsPCa was 100%, was taken as the protective factor, and the risk stratification model was finally proposed.Results:Of all 549 cases, 44 were CsPCa, 35 were clinically insignificant prostate cancer and 470 were non-prostate cancer. There were significant differences in age (71 vs. 67 years old), tPSA (11.95 vs. 8.75 ng/ml), PSAD [0.31 vs. 0.18 ng/(ml·cm 3)], f/tPSA (0.12 vs. 0.16) and DRE positive rate (38.6% vs. 7.3%) between CsPCa group and non-CsPCa group ( P<0.01). Cut-off values were taken in ROC analysis when the Youden index was at its maximum. The optimal cut-off values of each continuous variable were: age=65 years, tPSA=10ng/ml, f/tPSA=0.2 and PSAD=0.15 ng/(ml·cm 3). Multivariate analysis showed that ages over 65 years ( OR=3.43, 95% CI 1.55-7.58, P=0.002), f/t PSA ratio<0.2 ( OR=3.84, 95% CI 1.28-11.56, P=0.016), PSAD>0.15 ng/(ml·cm 3) ( OR=3.60, 95% CI 1.13-11.51, P=0.03) and positive DRE ( OR=5.20, 95% CI 2.39-11.32, P<0.001) were independent risk factors of CsPCa. When NPV was 100%, the cut-off values were taken as the protective factors: age≤55 years, f/tPSA≥0.3, PSAD≤0.1 ng/(ml·cm 3). Combined with independent risk factors, preliminary risk stratification was conducted: those with ≥2 high risk factors were considered as high risk group, those with ≥2 protective factors were considered as low risk group, and the middle region was considered as medium risk group. Conclusions:Patients with age>65 years, f/tPSA<0.2, PSAD > 0.15 ng/(ml·cm 3) and DRE positive are independent risk factors of CsPCa in mpMRI negative patients. Patients in the high-risk group were recommended to undergo prostate biopsy, while patients in the low-risk group could be considered to avoid biopsy.

In China's new healthcare reform, the pilot local governments explore the practice of establishing a new model of hierarchical diagnosis and treatment system.Zhejiang Province has adopted a special policy of effectively allocating hospital resources and human resources, and efficiently improving primary healthcare institution capability and patient satisfaction(hereinafter referred to as double allocation, double improvement), focusing on the implementation of the 'Healthcare Talents Project', in order to fill a vacancy of human resources in primary healthcare institutions.This paper uses system dynamics modeling and the WISN method of WHO to estimate the gap in physician supply in primary healthcare institutions.After building the system dynamics model of 'Healthcare Talents Project', this paper simulates the influence of the policy on the vacancy of doctors in primary healthcare institutions and analyzes the sensitivity of regulatory factors.The simulation results show that, there are a big gap in physician supply of about 14,000 to build the hierarchical diagnosis and treatment system.The project can gradually increase the number of primary doctors, and the policy may fill the vacancy by 2021.However, if the efficiency of the hospital doctors who give assistance to primary institutions is increased by 10%, the targeted training and recruitment 100% achieve the policy plans and objectives, the project goal may be achieved by 2020.Therefore, this project can effectively adjust the human resources structure quickly and reasonably, and it can be used as reference for the reform of hierarchical diagnosis and treatment system.

Objective To investigate the effect of galangin on proliferation and apoptosis of glioma cells in vitro.Methods (1) The glioma cells U87 and U25 1were divided into blank control group,DMSO group,100,200,300 and 400 μmol/L galangin treatment groups.MTT was used to study the effects of drugs on the proliferation of U251 and U87 cells.(2) Hoechest staining was used to observe cell apoptosis in the presence of different concentrations of galangin (0,100 and 200 μmol/L).(3) Flow cytometry was employed to detect the apoptosis of U251 and U87 cells in the presence of different concentrations of galangin (1 00 and 200 μmol/L).(4) Western blotting was employed to detect the expressions of apoptosis-related protein 3-Catenin,B-cell lymphoma-2 (Bcl-2),Bcl-2 related protein gene (Bax),cleaved-caspase-3,cleaved-caspase-9 and poly (ADP-ribose) polymerase (PARP) in the presence of different concentrations of galangin.Results (1) The proliferation of U251 and U87 cells was obviously inhibited atter 100,200,300 and 400 μmol/L galangin treatments,and dose-effect relation was noted.The concentrations of galangin at half rate of inhibition (IC50) were 281,321,276 and 229 μmol/L in U251 cells,and 289.4,261.1,247.4 and 225.3 μ mol/L in the U87 cells after 100,200,300 and 400μmol/L galangin treatments for 24 h.(2) Under the action of galangin,corresponding increase in apoptosis rates of U251 and U87 cells was noted following the increase of galangin concentrations (0,100 and 200 μmol/L),with significant differences (P＜0.05).(3) The detection of cell apoptosis by flow cytometry found similar changes.(4) Western blotting results indicated that galangin at the concentration of 0,100 and 200 μmol/L could significantly decrease the expressions of apoptosis-related protein 3-Catenin and Bcl-2,and increase the Bax,cleaved-caspase-3 and cleaved-caspase-9,and cleaved-PARP expressions;significant differences were noted between each two concentrations (P＜0.05).Conclusion Galangin can inhibit proliferation of glioma cells U251 and U87,and induce mitochondrial pathway of apoptosis via Wnt/β-Catenin signaling.

ObjectiveTo evaluate the value of intraoperative frozen section examination (IFS) in the diagnosis and surgical procedures selection for renal occupying lesions. MethodsFrom January 2006 to December 2010,IFS was used in 114 men and 81 women with renal occupying lesions.The mean age was 52 years (range 17 -78).In 104,89,and 2 patients,lesions were in the right,left and bilateral kidneys,respectively.All patients underwent physical examination,129 were asymptomatic at presentation while clinical symptoms were observed in 66.The largest dimension of the tumors were 4 cm or less in 128 patients,4- 7 cm in 49,and larger than 7 cm in 18,respectively.The outcomes between IFS and postoperative routine paraffin section examination were compared.In cases with renal tumors nephrectomy or partial nephrectomy was performed.The results of IFS were compared between the 2 groups. ResultsThe sensitivity and specificity of IFS for renal malignant lesions was 96.6％ and 100％,respectively.The total accuracy rate of IFS for renal occupying lesions was 97.4％.By subgroup analysis,the accuracy rate of clear cell carcinoma,papillary cell carcinoma,chromophobe cell carcinoma,sarcomatoid cancer,nephroblastoma,benign tumor and urothelial cancer was 94.3％,25.0％,16.7％,0,0,97.6％ and 100.0％,respectively.Partial nephrectomy and nephrectomy were performed in 57 and 123 patients with renal tumors,respectively.The surgical procedures selection was significantly associated with the lesion size (4 cm or less for 80.7％ vs 62.6％,P =0.015) and the malignant lesion diagnosed by IFS (31.6％ vs 93.5％,P＜0.001). Conclusion The accuracy of frozen section analysis for renal malignant lesions during surgery is reliable and significantly high,and the results can exert an important impact on surgical procedures selection.

BACKGROUND: Studies have demonstrated that incidence rate of acute rejection in renal transplant recipients with pre-production of major histocompatibility complex class I chain-related gene A (MICA), including parts of autoantibody, before transplantation in body, is obviously greater than that of recipients with negative antibody. OBJECTIVE: To investigate effects of exogenous antigen on MICA expression in endothelial cells. METHODS: The endothelial cells were cultured with exogenous recombinant MICA protein (group M5, M10 and M25) and heat shock protein-70 (group H5, H10 and H25) with dosages of 5, 10 and 25 μg/L, respectively, for 48 hours. Same volume of phosphate buffer saline was added into the control groups. RESULTS AND CONCLUSION: At 48 hours after induction, the expressions of MICA mRNA and protein were increased significantly in each experimental group (M5, M10 and M25) than that of the control group with significant (P < 0.05). The expression of MICA mRNA and MICA protein of group M5 and group M10 were remarkably higher than group M25 (P < 0.05); however, there was no significant difference between group M5 and M10 (P > 0.05). The expression of MICA membrane protein in the group M10 was obviously greater than that of the group M5 and M25 (P < 0.05). The level of soluble MICA (sMICA) in experimental groups (M5, M10 and M25) was decreased obviously comparing with that of the control group. These differences had statistical significances (P < 0.05). But there was no significant difference among the experimental groups (P > 0.05). However, the expression of MICA gene and sMICA level did not change after heat shock protein-70 stimulation. The exogenous MICA antigen up-regulates the expression of MICA mRNA and protein, especially increases the expression of membrane protein on the cell surface significantly, but sMICA in supernatant was dramatically decreased.