Background and purpose:Members of the serine protease inhibitor(SERPIN)family can influence tumorigenesis,progression,and prognosis by modulating processes such as apoptosis,invasion,metastasis,and angiogenesis in tumor cells.However,their role in pancreatic cancer remains unclear.This study aimed to investigate the impact of high expression of serine protease inhibitor A3(SERPINA3)on the proliferation,apoptosis,migration,and chemoresistance of pancreatic cancer cells and its mechanism.Methods:This study analyzed the SERPINA3 expression levels in the normal pancreatic ductal epithelial cell line hTERT-HPNE and pancreatic cancer cell lines SW1990,Capan-1,PANC-1,and ASPC-1 by real-time reverse transcription quantitative polymerase chain reaction(qRT-PCR).We established gemcitabine-resistant pancreatic cancer cell lines PANC-1/R and ASPC-1/R,and used qRT-PCR assay and cell counting kit-8(CCK-8)to determine the SERPINA3 expression levels in the constructed resistant cell lines and their parental sensitive cell lines,as well as the differences in their chemosensitivity to gemcitabine.We constructed the SERPINA3-knockdown cell line si-SERPINA with siRNA,and the negative control group si-SERPINA#NC with siRNA negative control.We used MDA assay,CCK-8 assay,EdU cell proliferation assay,transwell migration assay,matrigel invasion assay,scratch assay,and apoptotic assay to respectively detect the lipid oxidation levels,proliferation,migration,invasion,wound-healing ability,and the influence on apoptosis of the gemcitabine-resistant pancreatic cancer cells in the si-SERPINA group and the si-SERPINA#NC group.Results:Compared with normal pancreatic ductal epithelial cells hTERT-HPNE,the expression level of SERPINA3 in various pancreatic cancer cell lines was significantly increased(P＜0.05).mRNA and protein expression levels of SERPINA3 in PANC-1/R and ASPC-1/R were significantly increased compared with those in parent cells(P＜0.001).When SERPINA3 was knocked down in PANC-1/R and ASPC-1/R cells,the survival rate of the cells under different concentrations of gemcitabine chemotherapy decreased,and MDA detected that the lipid oxidation level was increased(P＜0.001).In addition,the proliferation rate of PANC-1/R and ASPC-1/R cell lines with SERPINA3 knockout,the number of migrating/invading cells and the healing rate of scratch test were significantly decreased(P＜0.01),and flow cytometry demonstrated that the number of apoptotic cells was increased(P＜0.05).These results suggest that SERPINA3 knockdown can inhibit the proliferation,migration,invasion and wound healing ability of gemcitabine-resistant pancreatic cancer cells,and promote the apoptosis of these resistant cells.Conclusion:SERPINA3 overexpression was found in various pancreatic cancer cells.SERPINA3 overexpression promoted malignant progression and chemotherapy resistance of pancreatic cancer,and interference with SERPINA3 expression promoted ferroptosis and enhanced chemotherapy sensitivity of gemcitabine-resistant pancreatic cancer cells.

Purpose To investigate the correlation between tumor budding in penile squamous cell carcinoma(SCC)and its clinicopathological features.Methods Clinical data and pathological slides from 69 cases of penile SCC were collected.Tumor budding was examined microscopically,and statistical analyses were performed to assess the relationship between tumor budding and various clinicopathological features.Results Among the 69 cases of pe-nile SCC,41 cases exhibited low-grade tumor budding and 28 cases displayed high-grade tumor budding.Tumor bud-ding was significantly correlated with tumor size,urethral invasion,invasion of the urethral corpus spongiosum,inva-sion of the penile corpus cavernosum,tumor necrosis,nerve invasion,vascular tumor thrombus,and pathological T stage(all P＜0.05).In contrast,no significant association was found between tumor budding and HPV-related versus non-HPV-related penile SCC(P＞0.05).Log-rank survival analysis indicated that patients with high-grade tumor bud-ding had a significantly lower survival rate compared to those with low-grade tumor budding(P＜0.05).Conclusion Tumor budding is a distinct pathological feature of penile SCC,and high-grade tumor budding is associated with a more aggressive biological behavior.

Objective To investigate the safety and efficacy of combined confluence microdissection or balloon dilatation followed by laparoscopic transcystic common bile duct exploration (LTCBDE) in day surgery laparoscopic cholecystectomy. Methods The clinical data of 203 patients with day surgery laparoscopic cholecystectomy combined with LTCBDE from June 2021 to June 2024 in Beijing Friendship Hospital, Capital Medical University were retrospectively analyzed. They were divided into an observation group (59 cases, including 42 cases of confluent microdissection and 17 cases of balloon dilatation) and a conventional group (144 cases) according to the surgical technique used. Baseline characteristics, intraoperative exploration results, operation time, postoperative recovery and complications were compared between the two groups. Results The history of preoperative cholangitis or pancreatitis (P<0.001) was more common in the observation group. And total bilirubin level was significantly higher in the observation group than in the conventional group (P=0.035). The observation group had a longer operative time (P=0.014) and higher hospitalization costs (P=0.001), but there was no difference in intraoperative bleeding and postoperative discharge time. There were no serious postoperative complications in either group. Conclusions Under the premise of strict patient screening, day surgery LTCBDE combined with confluence microdissection or balloon dilatation can effectively solve the problem of difficult choledochoscopic access, with high safety and thoroughness of exploration. Both techniques provide a feasible minimally observation solution for day surgery biliary exploration.

Background and purpose:Members of the serine protease inhibitor(SERPIN)family can influence tumorigenesis,progression,and prognosis by modulating processes such as apoptosis,invasion,metastasis,and angiogenesis in tumor cells.However,their role in pancreatic cancer remains unclear.This study aimed to investigate the impact of high expression of serine protease inhibitor A3(SERPINA3)on the proliferation,apoptosis,migration,and chemoresistance of pancreatic cancer cells and its mechanism.Methods:This study analyzed the SERPINA3 expression levels in the normal pancreatic ductal epithelial cell line hTERT-HPNE and pancreatic cancer cell lines SW1990,Capan-1,PANC-1,and ASPC-1 by real-time reverse transcription quantitative polymerase chain reaction(qRT-PCR).We established gemcitabine-resistant pancreatic cancer cell lines PANC-1/R and ASPC-1/R,and used qRT-PCR assay and cell counting kit-8(CCK-8)to determine the SERPINA3 expression levels in the constructed resistant cell lines and their parental sensitive cell lines,as well as the differences in their chemosensitivity to gemcitabine.We constructed the SERPINA3-knockdown cell line si-SERPINA with siRNA,and the negative control group si-SERPINA#NC with siRNA negative control.We used MDA assay,CCK-8 assay,EdU cell proliferation assay,transwell migration assay,matrigel invasion assay,scratch assay,and apoptotic assay to respectively detect the lipid oxidation levels,proliferation,migration,invasion,wound-healing ability,and the influence on apoptosis of the gemcitabine-resistant pancreatic cancer cells in the si-SERPINA group and the si-SERPINA#NC group.Results:Compared with normal pancreatic ductal epithelial cells hTERT-HPNE,the expression level of SERPINA3 in various pancreatic cancer cell lines was significantly increased(P＜0.05).mRNA and protein expression levels of SERPINA3 in PANC-1/R and ASPC-1/R were significantly increased compared with those in parent cells(P＜0.001).When SERPINA3 was knocked down in PANC-1/R and ASPC-1/R cells,the survival rate of the cells under different concentrations of gemcitabine chemotherapy decreased,and MDA detected that the lipid oxidation level was increased(P＜0.001).In addition,the proliferation rate of PANC-1/R and ASPC-1/R cell lines with SERPINA3 knockout,the number of migrating/invading cells and the healing rate of scratch test were significantly decreased(P＜0.01),and flow cytometry demonstrated that the number of apoptotic cells was increased(P＜0.05).These results suggest that SERPINA3 knockdown can inhibit the proliferation,migration,invasion and wound healing ability of gemcitabine-resistant pancreatic cancer cells,and promote the apoptosis of these resistant cells.Conclusion:SERPINA3 overexpression was found in various pancreatic cancer cells.SERPINA3 overexpression promoted malignant progression and chemotherapy resistance of pancreatic cancer,and interference with SERPINA3 expression promoted ferroptosis and enhanced chemotherapy sensitivity of gemcitabine-resistant pancreatic cancer cells.

Purpose To investigate the correlation between tumor budding in penile squamous cell carcinoma(SCC)and its clinicopathological features.Methods Clinical data and pathological slides from 69 cases of penile SCC were collected.Tumor budding was examined microscopically,and statistical analyses were performed to assess the relationship between tumor budding and various clinicopathological features.Results Among the 69 cases of pe-nile SCC,41 cases exhibited low-grade tumor budding and 28 cases displayed high-grade tumor budding.Tumor bud-ding was significantly correlated with tumor size,urethral invasion,invasion of the urethral corpus spongiosum,inva-sion of the penile corpus cavernosum,tumor necrosis,nerve invasion,vascular tumor thrombus,and pathological T stage(all P＜0.05).In contrast,no significant association was found between tumor budding and HPV-related versus non-HPV-related penile SCC(P＞0.05).Log-rank survival analysis indicated that patients with high-grade tumor bud-ding had a significantly lower survival rate compared to those with low-grade tumor budding(P＜0.05).Conclusion Tumor budding is a distinct pathological feature of penile SCC,and high-grade tumor budding is associated with a more aggressive biological behavior.

Objective To investigate the mechanism behind the protective effects of gastrodin against microglia-mediated inflammatory responses following hypoxic-ischemic brain damage(HIBD)in neonatal mice.Methods Thirty-six 10-day-old C57BL/6J mice were randomized into sham-operated group,HIBD(induced by ligation of the left common carotid artery followed by hypoxia for 40 min)group,and HIBD with gastrodin treatment groups(n=12).In gastrodin treatment group,100 mg/kg gastrodin was injected intraperitoneally 1 h before and at 2 and 12 h after hypoxia.After the treatments,the expressions of CCR5,AKT,p-AKT,and TNF-α and the co-expression of IBA1 and CCR5 in the corpus callosum of the mice were detected with Western blotting and immunofluorescence double staining.In a BV2 microglial cell model of oxygen-glucose deprivation(OGD),the effects of pretreatment with gastrodin and Maraviroc(an CCR5 antagonist)on protein expressions of CCR5,AKT,p-AKT,TNF-α and IL-1β were evaluated using Western blotting and immunofluorescence double staining.Results The neonatal mice with HIBD showed significantly increased expressions of CCR5 and TNF-α with lowered p-AKT expression in the brain tissues,and GAS treatment obviously reversed these changes.HIBD also significantly increased the co-expression of IBA1 and CCR5 in the corpus callosum of the mice,which was obviously lowered by gastrodin treatment.In BV2 cells,OGD significantly increased the expressions of CCR5,TNF-α,and IL-1β and decreased the expression of p-AKT,and these changes were inhibited by treatment with gastrodin,Maraviroc or their combination;the inhibitory effect of the combined treatment did not differ significantly from that of gastrodin or Maraviroc alone.Conclusion Gastrodin can produce neuroprotective effects in neonatal mice with HIBD by inhibiting inflammatory cytokine production and activate AKT phosphorylation via inhibiting CCR5.

Objective To investigate the mechanism behind the protective effects of gastrodin against microglia-mediated inflammatory responses following hypoxic-ischemic brain damage(HIBD)in neonatal mice.Methods Thirty-six 10-day-old C57BL/6J mice were randomized into sham-operated group,HIBD(induced by ligation of the left common carotid artery followed by hypoxia for 40 min)group,and HIBD with gastrodin treatment groups(n=12).In gastrodin treatment group,100 mg/kg gastrodin was injected intraperitoneally 1 h before and at 2 and 12 h after hypoxia.After the treatments,the expressions of CCR5,AKT,p-AKT,and TNF-α and the co-expression of IBA1 and CCR5 in the corpus callosum of the mice were detected with Western blotting and immunofluorescence double staining.In a BV2 microglial cell model of oxygen-glucose deprivation(OGD),the effects of pretreatment with gastrodin and Maraviroc(an CCR5 antagonist)on protein expressions of CCR5,AKT,p-AKT,TNF-α and IL-1β were evaluated using Western blotting and immunofluorescence double staining.Results The neonatal mice with HIBD showed significantly increased expressions of CCR5 and TNF-α with lowered p-AKT expression in the brain tissues,and GAS treatment obviously reversed these changes.HIBD also significantly increased the co-expression of IBA1 and CCR5 in the corpus callosum of the mice,which was obviously lowered by gastrodin treatment.In BV2 cells,OGD significantly increased the expressions of CCR5,TNF-α,and IL-1β and decreased the expression of p-AKT,and these changes were inhibited by treatment with gastrodin,Maraviroc or their combination;the inhibitory effect of the combined treatment did not differ significantly from that of gastrodin or Maraviroc alone.Conclusion Gastrodin can produce neuroprotective effects in neonatal mice with HIBD by inhibiting inflammatory cytokine production and activate AKT phosphorylation via inhibiting CCR5.

Objective:To investigate the clinical effects of free posterior interosseous artery perforator flap on reconstruction of the soft tissue defects of multiple fingers in one hand.Methods:Clinical data of 9 patients with soft tissue defect of multiple fingers in one hand admitted to the Department of Hand Surgery, Yongkang Orthopedic Hospital between January 2021 and August 2023 were retrospectively studied. The patients were 7 males and 2 females, aged between 19 and 55 years old. The soft tissue defects of 2 patients with 3 fingers injury and 7 patients with 2 fingers injury were reconstructed with free posterior interosseous artery perforator flaps. The size of defects in single finger was 1.5 cm×2.5 cm-3.0 cm×4.5 cm. The size of flap was 2.5 cm×7.0 cm-3.5 cm×13.0 cm. The posterior cutaneous nerve of the forearm were sutured to the intrinsic nerve of palmar side of the 14 fingers. Donor sites in forearm were directly sutured. After discharge, regular follow-up at outpatient clinic and through WeChat interviews were conducted to observe the appearance and texture of the flaps, finger fullness, donor site function and appearance, as well as patient satisfaction. For the flaps with anastomosis of cutaneous nerve, the recovery of TPD of the flap was tested. Sensory recovery was assessed according to the British Medical Research Council (BMRC) sensory function assessment criteria. Hand functions were evaluated using the Total Active Movement (TAM) of fingers of the Hand Surgery of the Chinese Medical Association.Results:All of the 20 flaps survived and were evaluated through follow-up visits, which ranged from 6 to 25 months with an average of 13 months. The flaps were soft, unbloated and without obvious pigmentation. The protective sensations were recovered with an average static TPD of 14 flaps anastomosed with cutaneous nerve was of 9.7 (7-13) mm. Among them, S 3+ 10 fingers, S 3 4 fingers; 6 fingers of flap without sutured cutaneous nerve, S 2 5 fingers, S 1 1 finger. According to the TAM of Hand Surgery of Chinese Medical Association, the results were excellent for 15 fingers and good for 5 fingers. There was no scar contracture in the forearm donor sites and motor function was not affected. Conclusion:Posterior interosseous artery perforator flap has the advantages of thin flap, similar texture to the hand, good recovery in sensory, and minimal damage to the donor site. Therefore, it is a good option in reconstruction of soft tissue defects of multiple fingers in one hand.

Objectives:To analyze the clinical efficacy of Cheneau brace in adolescent idiopathic scoliosis(AIS)patients and to discuss the changes of curve type during the treatment.Methods:75 patients with AIS treated with Cheneau brace from October 2016 to November 2021 were included.There were 69 females and 6 males,averaged 11.9±1.0 years old when starting to wear the brace,with a mean initial main curve Cobb angle of 29.8°±8.5° and an initial Risser's sign ≤2.Clinical and imaging data were collected,including age,gender,menarche age,time of daily wear,and full-length radiographs of the spine before initial treatment and at the final follow-up.All the patients were followed up for 1 year after finishing brace treatment.The curve types of the patients were counted using the main curve type and the modified Lenke classification system(mLenke),and changes in curve types and the efficacy of bracing in patients with different curve types were analyzed.Results:After 25.7±10.8 months of brace treatment,the patients averaged 14.2±1.2 years old when finishing brace treatment,and the mean main Cobb angle was 27.6°±12.0°.Among the patients,scoliosis was controlled in 63 while progressed in 12,and 23 patients were treated surgically.The rates of success of brace and control of curve were higher in patients with main lumbar curve than those in patients with main thoracic curve(P=0.043,P=0.003).There was a difference in the rate of brace success and control of curve in patients with each mLenke classification(P＜0.001,P=0.005).The highest rates of brace success and control of curve were found in patients of mLenke type Ⅴ,while mLenke type Ⅳ patients had the lowest success rate of brace treatment success and mLenke type Ⅱ patients had the lowest rate of control of curve.Before and after brace treatment,there was a difference in the proportion of patients with each main curve type(P=0.019),and the proportion of patients with each mLenke classification had no statistical difference(P=0.071).There were differences in the rate of change of curve type in each main curve type and each mLenke classification both(P＜0.001,P=0.020).No significant differences were seen in the rates of success of brace treatment and control of curve between patients with and without change in curve type under the two curve type classifications.Conclusions:The efficacy of Cheneau brace varies among AIS patients with different curve types.Patients with a main thoracic curve tend to have a poor response to bracing compared with patients of main lumbar curve;During the treatment with Cheneau brace,the patient's curve types may change,and the overall trend of curve type change is upward shifting of the main curve.The rate and outcome of curve type changes vary among patients with different curve types,and curve type changes may also occur in curve controlled or corrected patients.

Objective:To investigate the influencing factors for textbook outcome (TO) after hepatectomy for hepatolithiasis.Methods:The retrospective case-control study was conducted. The clinicopathological data of 216 patients with hepatolithiasis who were admitted to The First Affi-liated Hospital of Anhui Medical University from January 2015 to March 2023 were collected. There were 69 males and 147 females, aged 61(range, 22-85)years. Observation indicators: (1) treatment situations; (2) TO after hepatectomy; (3) Influencing factors for TO after hepatectomy. Measurement data with normal distribution were represented as Mean± SD. Measurement data with skewed distribution were represented as M(range). Count data were expressed as absolute numbers. Logistic regression models were used for univariate and multifactorial analyses. Results:(1) Treatment situations. All the 216 patients underwent hepatectomy, including 45 cases of laparoscopic hepatectomies and 171 cases of open hepatectomies, 161 cases of anatomical hepatectomies and 55 cases of non-anatomical hepatectomies. All the 216 patients underwent intraoperative choledochoscopy exploration and lithotripsy. There were 170 patients with normal Oddi sphincter function and 46 patients with Oddi sphincter dysfunction. All the 216 patients underwent biliary drainage, including 198 cases of external T-tube drainage and 18 cases of internal bile-intestinal drainage. The operation time was (226±75)minutes and volume of intraoperative blood loss was (106±82)mL. There were 29 patients with perioperative blood transfusion and 14 patients with intraoperative severe adverse events. There were 189 patients achieved immediate stone clearance. Of 183 patients with intraoperative bile cultures, 76 cases were positive for bacteria culture. (2) TO after hepatectomy. Of 216 patients, 93 cases had postoperative complications, all of which were successfully discharged after active treatment. One patient had surgery-related death within 90 days after surgery, and the cause of death was liver failure. Five patients were readmitted within 90 days after surgery, and 18 patients had postoperative stone residual. Of 216 patients, 164 cases achieved TO postoperatively and 52 cases did not achieve TO postoperatively. (3) Influencing factors for TO after hepatectomy. Results of multivariate analysis showed that cholangitis, stone distribution, surgical approaches, anatomical hepatectomy, immediate stone removal and postoperative review of choledochoscopy were independent influencing factors for TO after hepatectomy in patients with hepatolithiasis ( P<0.05). Conclusion:Cholangitis, stone distribution, surgical approaches, anatomical hepatectomy, imme-diate stone removal and postoperative review of choledochoscopy are independent influencing factors for TO after hepatectomy in patients with hepatolithiasis.