Objective:To investigate the regulatory effect of transient receptor potential cation channel subfamily C member 3 (TRPC3) on the retina in oxygen-induced retinopathy (OIR) mice and biological behavior of human retinal vascular endothelial cells (HREC).Methods:A total of 32 healthy SPF grade 7-day-old C57BL/6 mice were selected and randomly divided into a control group and an OIR group by the random number table method, with 16 mice in each group.The control group received no special treatment, and the OIR model was established in the OIR group.On postnatal day 17 (PN17), the success of the model establishment was verified by immunofluorescence staining of the retinal patch.The in vitro cultured HREC were divided into a normal control group, a transfection reagent group, and a si-TRPC3 group.The normal control group received no special treatment, while the transfection reagent group and the si-TRPC3 group were transfected with transfection reagent or transfection reagent + si-TRPC3.The relative expression of TRPC3 mRNA was detected by real-time quantitative fluorescence PCR.The relative expressions of TRPC3, transcription factor NF-E2 related factor (Nrf2), and superoxide dismutase (SOD) proteins were determined by Western blot.HREC were further divided into a normal control group, a vascular endothelial growth factor (VEGF) group, a si-TRPC3 group, and a Pyr3 (TRPC3 channel inhibitor) group, which were cultured in complete medium, medium containing 20 ng/ml VEGF recombinant protein, medium containing 20 ng/ml VEGF recombinant protein (si-TRPC3 transfection for 72 hours), and medium containing 20 ng/ml VEGF recombinant protein+ 1 μmol/L Pyr3 for 48 hours, respectively.The proliferation ability of HREC was detected using cell counting kit 8 (CCK-8). The horizontal and vertical migration ability of cells were detected by cell scratch assay and transwell assay, respectively.This study followed the 3R principles of animal welfare and was approved by the Ethics Committee of Hebei Eye Hospital (No.2023LW04). Results:Pathological neovascular clusters with strong fluorescent staining appeared in the retina of OIR mice on PN17.The relative expressions of TRPC3 mRNA and protein in the retina of OIR mice were 2.057±0.244 and 1.517±0.290, respectively, significantly higher than 0.983±0.033 and 0.874±0.052 of control group ( t=6.165, 3.094; both at P<0.05). The relative expression levels of TRPC3 mRNA and protein were significantly lower, and the relative expression levels of Nrf2 and SOD proteins were higher in the si-TRPC3 group than in the normal control and transfection reagent groups, and the differences were statistically significant (all at P<0.05). The CCK-8 experiment results showed that the cell absorbance value was higher in the VEGF group than in the normal control group, and lower in the si-TRPC3 and Pyr3 groups than in the VEGF group, with statistically significant differences (all at P<0.05). The results of the cell scratch experiment showed that the lateral migration rate of VEGF group cells was higher than that of normal control group, while the lateral migration rate of si-TRPC3 group and Pyr3 group cells was lower than that of VEGF group, and the differences were statistically significant (all at P<0.05). The transwell experiment results showed that the number of stained cells in the VEGF group was higher than that in the normal control group, and the number of stained cells in the si-TRPC3 group and Pyr3 group was lower than that in the VEGF group, with statistically significant differences (all at P<0.05). Conclusions:Hypoxia induces increased TRPC3 expression in OIR mouse retina, and downregulation of TRPC3 inhibits HREC proliferation and migration.The mechanism is related to the activation of the Nrf2-related oxidative stress pathway.

Objective To explore the action mechanism of tauroursodeoxycholic acid(TUDCA)promoting intracellular clearance of Burkholderia pseudomallei(B.pseudomallei)in RAW264.7 macrophages.Methods After TUDCA of different concentrations were used to treat RAW264.7 cells pre-infected with B.pseudomallei for 8 h or not,flow cytometry was applied to detect the apoptosis of the infected and control cells.In addition,another endoplasmic reticulum stress(ERS)inhibitor 4-PBA was used to detect the apoptosis and proliferation of host cells after B.pseudomallei infection with Annexin-V/PI double staining and MTT cell proliferation assay.Furthermore,after transfected with CHOP siRNA,Western blotting and flow cytometry were employed to detect the effect of TUDCA on the expression levels of Caspase-3 and Caspase-12 and the changes in apoptotic rate after B.pseudomallei infection,respectively.Finally,the effect of TUDCA on intracellular multiplication of infected RAW264.7 cells were observed to estimate the CFU value in the presence and absence of CHOP siRNA.Results Under different concentrations of TUDCA,100 or 200 μmol/L TUDCA significantly reduced B.pseudomallei-induced apoptosis in RAW264.7 cells(P＜0.05).Meanwhile,both TUDCA and 4-PBA treatment could decrease the apoptosis induced by B.pseudomallei infection by ERS(P＜0.05).Further,the expression levels of Caspase-3 and Caspase-12 were obviously increased after B.pseudomallei infection compared with uninfected groups,but their expression levels in the siCHOP group was significantly lower than that in the siC group.Besides,flow cytometry also showed that TUDCA could reduce apoptosis induced by B.pseudomallei infection(P＜0.05),but no significant effect of TUDCA on apoptosis was observed under CHOP knockdown.Finally,intracellular CFU assay indicated that TUDCA treatment promoted the host cell clearance of B.pseudomallei(P＜0.05),but no such effect was observed in siCHOP group.Conclusion In B.pseudomallei infected RAW264.7 cells,TUDCA promotes the intracellular clearance of the bacteria by inhibiting ERS-induced apoptosis.

Objective:To investigate the risk factors of urethrovesical anastomotic leakage after laparoscopic radical prostatectomy.Methods:The clinical data of 292 patients who underwent laparoscopic radical prostatectomy in the Tenth People's Hospital Affiliated to Tongji University from January to December 2021 were retrospectively analyzed. According to whether there was anastomotic leakage, the patients were divided into leakage group (27 cases) and non-leakage group (265 cases). There were no significant differences in age [(71.5±6.5) years vs. (70.2±6.4) years], body mass index [(24.5±3.6) kg/m 2 vs. (24.2±3.0) kg/m 2], prostate volume[40(27.3, 63.2)ml vs. 38(28.1, 56.2)ml], Gleason score, clinical stage, and risk classification between the leakage group and the non-leakage group ( P>0.05), but the total prostate-specific antigen in the leakage group was significantly higher than that in the non-leakage group[20.0 (9.6, 79.0) ng/ml vs. 13.7 (8.5, 25.0) ng/ml, P=0.049]. Propensity score matching (PSM) was used to match the above indicators between the leakage group and the non-leakage group as 1∶1, so that the baseline of the two groups was balanced. The perioperative indicators of the matched two groups of patients were compared and analyzed. Statistically significant indicators were selected and included in univariate and multivariate logistic regression to analyze the risk factors of anastomotic leakage after radical prostatectomy. Finally, the receiver operating characteristic (ROC) curve was drawn, and the area under the curve (AUC) was calculated. The accuracy of each factor in predicting urine leakage was obtained. Results:After PSM, 24 cases were successfully matched. The leakage group had shorter membranous urethral length (MUL) [(15.5±2.2)mm vs. (17.5±1.5)mm, P<0.001], thinner membranous urethral wall thickness (UWT) [(9.5±1.9)mm vs. (10.6±1.5)mm, P=0.024], longer anastomotic time of urethrovesical neck[(21.6±4.1)min vs. (16.9±2.9)min, P<0.001] and higher failure rate of water injection test [16.7% (4/24) vs. 4.2% (1/24), P=0.045] than the non-leakage group. There was no significant difference in other indicators between the two groups. The results of multivariate logistic regression analysis showed that short MUL ( OR=0.544, 95% CI 0.335-0.884, P=0.014), narrow UWT ( OR=0.538, 95% CI 0.313-0.924, P=0.025) and long anastomotic time of urethrovesical neck ( OR=1.519, 95% CI 1.122-2.110, P=0.009) were independent risk factors for anastomotic urine leakage. ROC curve analysis showed that the AUC of MUL, UWT, and anastomotic time were 0.789 (95% CI 0.651-0.927), 0.715 (95% CI 0.562-0.868), and 0.842 (95% CI 0.731-0.953), respectively. Conclusions:Narrow and short membranous urethra and long anastomosis time in patients with laparoscopic radical prostatectomy may be independent risk factors for postoperative anastomotic leakage, which may predict the occurrence of anastomotic leakage.

OBJECTIVE: To discover the factors that may affect the use of selective tracheostomy among patients who have undergone head and neck surgeries with free flap reconstruction, so that the patients will not need tracheostomy nor receive the unnecessary treatment. METHODS: Five hundred and thirty-three patients who had undergone head and neck surgery with free flap reconstruction operated by the same team of surgery at Department of Oral and Maxillofacial Surgery at Peking University School of Stomatology from 2015 to 2016 were reviewed. Three hundred and twenty-one (60.2%) of these patients underwent selective tracheostomy. All the patients' demographic information, operation-related information, prior treatments, comorbidities and complications were recorded and analyzed. RESULTS: The patients with defects of the tongue, mouth floor, oropharynx and bilateral mandible, who underwent neck dissection and with previous radiotherapy and smoking habit were more likely to get selective tracheostomy. Usage of bulky soft tissue flap might also add to the risk of airway obstruction and the need of selective tracheostomy, while other factors were not significantly related to the risk of postoperative airway obstruction and the patients could be kept safe without selective tracheostomy. Most cases without tracheostomy were kept safe except one case, while 8.39% of the patients with tracheostomy suffered from tracheostomy related complications, mainly pneumonia and hemorrhage of the tracheostomy wound, yet none led to serious consequences or even death. CONCLUSION Selective tracheostomy is not necessary for patients who have undergone head and neck surgeries with free flap reconstruction except that there are defects at the tongue, oropharynx and mandible. Neck dissection, bulky soft tissue flap reconstruction, previous radiotherapy and smoking habit may also add to the risk of postoperative airway obstruction, while a favorable decision would involve a combination of all the above factors to assure the safety of the postoperative airway for the patients undergone head and neck surgeries with free flap reconstruction.
Airway Obstruction/surgery* ; Free Tissue Flaps ; Head and Neck Neoplasms/surgery* ; Humans ; Postoperative Complications/surgery* ; Reconstructive Surgical Procedures/adverse effects* ; Retrospective Studies ; Tracheostomy

Objective: To investigate the distribution of HIV-1 genetic subtypes and pretreatment drug resistance (PDR) among men who have sex with men (MSM) from 19 cities of 6 provinces in China. Methods: From April to November 2019, 574 plasma samples of ART-naive HIV-1 infected MSM were collected from 19 cities in Hebei, Shandong, Jiangsu, Zhejiang, Fujian, and Guangdong provinces, total ribonucleic acid (RNA) was extracted and amplified the HIV-1 pol gene region by nested polymerase chain reaction (PCR) after reverse transcription. Then sequences were used to construct a phylogenetic tree to determine genetic subtypes and submitted to the Stanford drug resistance database for drug resistance analysis. Results: A total of 479 samples were successfully amplified by PCR. The HIV-1 genetic subtypes included CRF01_AE, CRF07_BC, B, CRF55_01B, CRF59_01B, CRF65_cpx, CRF103_01B, CRF67_01B, CRF68_01B and unrecognized subtype, which accounted for 43.4%, 36.3%, 6.3%, 5.9%, 0.8%, 0.8%, 0.4%, 0.4%, 0.2% and 5.5%, respectively. The distribution of genetic subtypes among provinces is statistically different (χ²=44.141, P<0.001). The overall PDR rate was 4.6% (22/479), the drug resistance rate of non-nucleoside reverse transcriptase inhibitors, nucleoside reverse transcriptase inhibitors, and protease inhibitors were 3.5% (17/479), 0.8% (4/479) and 0.2% (1/479), respectively. The PDR rate of recent infections was significantly higher than that of long-term infections (χ²=4.634, P=0.031). Conclusions: The HIV-1 genetic subtypes among MSM infected with HIV-1 from 19 cities of 6 provinces in China are diverse, and the distribution of subtypes is different among provinces. The overall PDR rate is low, while the PDR rate of recent infections was significantly higher than that of long-term infections, suggesting the surveillance of PDR in recent infections should be strengthened.
China/epidemiology* ; Cities ; Drug Resistance ; Drug Resistance, Viral/genetics* ; Female ; Genotype ; HIV Infections/epidemiology* ; HIV Seropositivity/drug therapy* ; HIV-1/genetics* ; Homosexuality, Male ; Humans ; Male ; Phylogeny ; Reverse Transcriptase Inhibitors/therapeutic use* ; Sexual and Gender Minorities

OBJECTIVE: To compare clinical efficacy of intramedullary nail fixation following two-step closed reduction or limited open reduction for femoral subtrochanteric fractures. METHODS: Forty six patients with femoral subtrochanteric fractures were analyzed retrospectively from January 2014 to April 2020. Twenty four patients which including 16 males and 8 females, aged from 34 to 91 years old with an average of (55.42±18.25) years old, were treated with two step closed reduction and intramedullary nail fixation on supine position. Twenty-two patients which including 15 males and 7 females, aged from 33 to 87 years old with an average of (56.31±14.77) years old, were performed limited open reductionand intramedullary nail fixation. Operation time, intraoperative blood loss, complications and fracture healing time were recorded and compared between two groups. Postoperative Harris hip score at 8 months was applied to evalaute joint function. RESULTS: All patients were successfully complete operation without incision infection. All patients were followed up from 8 to 36 months with an average of (18.2± 6.1) months. Introperation blood loss, operation time in closed reduction group were (157.92±51.07) ml, (82.08±13.43) min respectively, while in limited open reduction group were (230.91±87.88) ml, (92.73±12.79) min respectively; while there were statistical difference between two groups ( CONCLUSION Femoral subtrochanteric fractures could be effectively treated by both methods. Two-step closed reduction and intramedullary nail fixation may be more advantageous in less tissue damage, shorter operation time and less intraoperative bleeding.
Adult ; Aged ; Aged, 80 and over ; Bone Nails ; Female ; Femoral Fractures ; Fracture Fixation, Intramedullary ; Fracture Healing ; Hip Fractures/surgery* ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome

OBJECTIVE: To study the prognosis and complications of proximal femoral nail antirotation(PFNA) in the treatment of elderly intertrochanteric fracture. METHODS: From January 2016 to November 2019, 127 cases of femoral intertrochanteric fracture were treated with PFNA, including 51 males and 76 females. The average age was 81.39±8.16 (range from 60 to 98). According to Evans classification, the numbers of cases of typeⅠa, typeⅠb, typeⅠc, typeⅠd and typeⅡwere 10, 46, 48, 16 and 7, respectively. Based on the AO /OTA classification, there were 10 cases of type 31-A1.2, 84 of type 31- A1.3, 17 of type 31-A2.2, 9 of type 31-A2.3, and 7 of type 31-A3. Preoperative and the final follow up, function of hip joint of patients treatedwith the PFNA were evaluated by harris hip score, and the postoperative complications were observed. RESULTS: No incision infection occurred in all patients after PFNA. In this research, 4 cases failed in internal fixation, three of them underwent artificial femoral head replacement;5 cases died within one year after operation;2 cases had multiple embolizations of pulmonary artery branches, which recovered after anticoagulation treatment. There were 48 cases complicated with pneumonia, among which 9 cases had pulmonary inflammation before operation;27 cases with pleural effusion, 3 cases with acute heart failure, 3 cases with acute renal insufficiency, except one case died of pneumonia, pleural effusion, hypoproteinemia and respiratory failure during hospitalization, the other patients recovered after treatment. All 119 patients were followed up for 6 to 36 months, with an average of (17.01±6.03) months, Harris hip score increased from 8.96±5.40 preoperation to 83.57±8.92 at the final follow-up ( CONCLUSION PFNA is a recommended option for the treatment of senile patients with intertrochanteric fracture. However, there were a lot of complications when femoral intertrochanteric fractures happened in aged patients, especially prevention and treatment of pulmonary embolism, promptly corrected low hemoglobin and low albumin, and reduce complications.
Aged ; Aged, 80 and over ; Bone Nails ; Female ; Fracture Fixation, Intramedullary/adverse effects* ; Hip Fractures/surgery* ; Humans ; Male ; Retrospective Studies ; Treatment Outcome

Objective To employ Logistic regression modeling to analyze the related factors influencing the accuracy of the high performance liquid chromatography (HPLC) determination of preservatives in beverages. Methods The HPLC separation was performed on a Zorbax Eclipse Plus C18 column with methanol-ammonium acetate solution as mobile phase. The external standard method was used to determine 5 beverage preservatives. The influencing factors on the measurement accuracy were statistically evaluated with univariate and multivariate analysis. Results Univariate analysis showed that the recovery rate of the added standard in the determination of coffee beverage preservatives by HPLC was affected by the pretreatment method, and the difference was statistically significant (P<0.05). Multivariate analysis showed that the main influencing factors on the accuracy of determination of sorbic acid was the pretreatment method (OR=5.406, P<0.05), while the sample type was a protective factor (OR=0.134, P<0.05). For the determination of benzoic acid, the main factor influencing the accuracy was the sample type (OR=1.112, P<0.05), while the pretreatment method was a protective factor (OR=0.447, P<0.05). Conclusion Logistic regression analysis can identify risk factors for the accuracy of the determination, and provide statistical modeling support for the experimental optimization.

OBJECTIVE: To investigate the method and clinical effects of arthroscopic rotator cuff repair with footprint ending shift for massive rotator cuff tear. METHODS: From March 2015 to April 2019, 27 patients with massive rotator cuff tear underwent arthroscopic repair, including 12 males and 15 females, ranging in age from 37 to 74 years old, with an average age of (56.1±8.9) years. According to the tension of rotator cuff, 5 patients were treated with double row suture bridge technique, and 22 patients were treated with single row technique, among which 7 patients underwent partial rotator cuff repair. Before and after operation, University of California Los Angeles(UCLA) score, American Shoulder and Elbow Surgeons (ASES) score and visual analogue scale (VAS) were used to evaluate shoulder joint function. RESULTS: All the patients were followed up, and the duration ranged from 8 to 40 months, with an average of (18.0±5.9) months. The UCLA score increased from preoperative 8.67±0.78 to final follow-up 30.89±1.07( CONCLUSION Arthroscopic rotator cuff repair with footprint ending shift for massive rotator cuff tear has satisfactory results, and single-row can be chosen to completely or partial repair rotator cuff tear for most patients.
Adult ; Aged ; Arthroscopy ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Range of Motion, Articular ; Rotator Cuff/surgery* ; Rotator Cuff Injuries/surgery* ; Treatment Outcome

BACKGROUND: Homozygous or compound heterozygous mutations in high temperature requirement serine peptidase A1 (HTRA1) gene are responsible for cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy (CARASIL). Recently, increasing evidence has shown that heterozygous HTRA1 mutations are also associated with cerebral small vessel disease (CSVD) with an autosomal dominant pattern of inheritance. This study was aimed to analyze the genetic and clinical characteristics of HTRA1-related autosomal dominant CSVD. METHODS: We presented three new Chinese cases of familial CSVD with heterozygous HTRA1 mutations and reviewed all clinical case reports and articles on HTRA1-related autosomal dominant CSVD included in PUBMED by the end of March 1, 2020. CARASIL probands with genetic diagnosis reported to date were also reviewed. The genetic and clinical characteristics of HTRA1-related autosomal dominant CSVD were summarized and analyzed by comparing with CARASIL. RESULTS: Forty-four HTRA1-related autosomal dominant CSVD probands and 22 CARASIL probands were included. Compared with typical CARASIL, HTRA1-related autosomal dominant probands has a higher proportion of vascular risk factors (P < 0.001), a later onset age (P < 0.001), and a relatively slower clinical progression. Alopecia and spondylosis can be observed, but less than those in the typical CARASIL. Thirty-five heterozygous mutations in HTRA1 were reported, most of which were missense mutations. Amino acids located close to amino acids 250-300 were most frequently affected, followed by these located near 150∼200. While amino acids 250∼300 were also the most frequently affected region in CARASIL patients, fewer mutations precede the 200th amino acids were detected, especially in the Kazal-type serine protease domain. CONCLUSIONS HTRA1-related autosomal dominant CSVD is present as a mild phenotype of CARASIL. The trend of regional concentration of mutation sites may be related to the concentration of key sites in these regions which are responsible for pathogenesis of HTRA1-related autosomal dominant CSVD.
Cerebral Infarction ; Cerebral Small Vessel Diseases/genetics* ; Heterozygote ; High-Temperature Requirement A Serine Peptidase 1/genetics* ; Humans ; Leukoencephalopathies/genetics* ; Mutation/genetics*