1.Inhibition of NLRP3 inflammasome activation improves PCOS granulosa cell apoptosis by regulating autophagy
Shanhua FU ; Lili BAO ; Da ZHAO ; Jun LI ; Fangting LIN ; Rong HU
Chinese Journal of Immunology 2024;40(8):1646-1652
		                        		
		                        			
		                        			Objective:To investigate the expression of NOD-like receptor protein 3(NLRP3)inflammasome and its relation-ship with granulosa cell apoptosis in patients with polycystic ovary syndrome(PCOS).Methods:Follicular fluid and ovarian granulosa cells were collected from 17 PCOS patients(PCOS group)and 20 non-PCOS patients(control group),and the expression levels of pro-inflammatory factor,TNF-α,IL-1β and IL-18 in follicular fluid were detected by ELISA.RT-PCR and Western blot were used to de-tect NLRP3 mRNA and NLRP3 inflammasome associated protein NLRP3,ASC and cleaved caspase-1 and autophagy related protein LC3-Ⅱ/LC3-Ⅰ and p62 expression level in granulosa cells.TUNEL assay was used to detect the apoptotic level of granulosa cells in both groups.NLRP3 silenced siRNA(si-NLRP3)and negative control sequence(si-NC)were transfected into human ovarian cancer granulosa cell line KGN cells by siRNA interference technique in vitro.TNF-α was used to simulate PCOS-related cell damage.KGN cells were divided into 4 groups according to different treatment methods:Ctrl group,TNF-α group,TNF-α+si-NLRP3 group and TNF-α+si-NC group.The levels of DHEA,testosterone,IL-1β and IL-18 in supernatant were detected by ELISA.TUNEL assay was used to detect the apoptosis level of KGN cells.The LC3-Ⅱ/LC3-Ⅰ and p62,NLRP3,ASC and cleaved caspase-1 protein expression levels,and NF-κB p-p65 level(NF-κB p-p65/NF-κB p65)in KGN cells were detected by Western blot.Results:Compared with con-trol group,the concentration of TNF-α,IL-1β,and IL-18 in follicular fluid of PCOS patients,and LC3-Ⅱ/LC3-Ⅰ,NLRP3 mRNA and the protein expression of NLRP3,ASC,and cleaved caspase-1 in granulosa cells of and apoptosis were significantly increased(P<0.05 or P<0.01),while the protein expression of p62 was significantly decreased(P<0.01).Compared with Ctrl group,the levels of DHEA,testosterone,IL-1β and IL-18 in supernatant of TNF-α group,TNF-α+si-NC group,and TNF-α+si-NLRP3 group,and LC3-Ⅱ/LC3-Ⅰ,ASC and cleaved caspase-1 protein expression,and the level of NF-κB p-p65 and apoptosis were significantly increased(P<0.05 or P<0.01),while p62 protein was significantly decreased(P<0.01).NLRP3 was significantly decreased except in TNF-α+si-NLRP3 group(P<0.01),while in the TNF-α group and TNF-α+si-NC group was significantly increased(P<0.01).However,com-pared with TNF-α group,the above indexes in TNF-α+si-NLRP3 group were significantly decreased(P<0.05),and TNF-α+si-NC group had no significant change(P>0.05).Conclusion:The over-activated NLRP3 inflammasome in granulosa cells may promote cell inflammatory injury and autophagy apoptosis in PCOS patients through NF-κB pathway.
		                        		
		                        		
		                        		
		                        	
2.Pancreatic glucagonoma with metastasis followed up for 15 years: A case report
Chen HAN ; Shanmei SHEN ; Min XIE ; Hong HUANG ; Shanhua BAO ; Feng WANG ; Yan BI ; Dalong ZHU
Chinese Journal of Endocrinology and Metabolism 2023;39(7):621-624
		                        		
		                        			
		                        			To explore the clinical characteristics, diagnosis, and management of pancreatic glucagonoma, a retrospective analysis of the clinical data and diagnostic algorithm of a patient with pancreatic glucagonoma was conducted, along with literature review. Pancreatic glucagonoma is a rare neuroendocrine tumor that originates from the pancreatic alpha cells. The main manifestations of glucagonoma syndrome(GS) include necrolytic migratory erythema, diabetes, anemia, and other systemic involvement. Early diagnosis of GS is challenging and crucial. Early identification and recognition of skin lesions contribute to timely diagnosis and treatment of the disease. Surgical resection is an effective treatment modality for glucagonoma.
		                        		
		                        		
		                        		
		                        	
3.Total pancreatectomy in treatment of pancreatic diseases: a single center experience
Gang LI ; Zhenghua CAI ; Chenglin LU ; Yinyin FAN ; Yudong QIU ; Shanhua BAO
Chinese Journal of Hepatobiliary Surgery 2021;27(6):407-410
		                        		
		                        			
		                        			Objective:To review our clinical experience on total pancreatectomy in treatment of pancreatic diseases.Methods:The clinical data of 25 patients with pancreatic diseases who underwent total pancreatectomy at Nanjing Drum Tower Hospital from February 2016 to October 2019 were retrospectively analyzed. The data on general information, surgical methods, postoperative complications, changes in postoperative pancreatic endocrine and exocrine functions and quality of life were analyzed.Results:There were 16 males and 9 females, aged (60.9±9.4) years. Planned selective surgery was carried out in 16 patients while unplanned operations in 9 patients. There were 15 patients with pancreatic neoplasms with 4 patients who received neoadjuvant or conversion therapy, 6 patients with intraductal papillary mucinous neoplasms, 3 patients with chronic pancreatitis and 1 patient with pancreatic neuroendocrine carcinoma. The total postoperative complication rate was 36% (9/25). One patient died in the perioperative period, and one patient underwent a second operation. After 6 months of operation, 2 of 24 patients had died. The remaining patients had a fasting blood sugar of (8.9±1.6) mmol/L, with an insulin dosage of (30.7±10.6) U/d. The average dosage of trypsin was (1.1±0.3) g/d. The quality of life score reached or exceeded the preoperative levels.Conclusions:Total pancreatectomy was safe and feasible for some pancreatic diseases. For patients with pancreatic cancer, its long-term oncological outcomes need further studies.
		                        		
		                        		
		                        		
		                        	
4.Analysis of influencing factors for pancreatic endocrine and exocrine insufficiency after pancreaticoduodenectomy
Zhenghua CAI ; Gang LI ; Shanhua BAO ; Xiaojie BIAN ; Yinyin FAN ; Xiaoyuan CHEN ; Yudong QIU
Chinese Journal of Digestive Surgery 2020;19(4):414-420
		                        		
		                        			
		                        			Objective:To investigate the influencing factors for pancreatic endocrine and exocrine insufficiency after pancreaticoduodenectomy.Methods:The retrospective case-control study was conducted. The clinicopathological data of 168 patients who underwent pancreaticoduodenectomy in the Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School from January 2016 to December 2017 were collected. There were 96 males and 72 females, aged (64±13)years, with a range from 38 to 75 years. Of the 168 patients, 36 had pancreatic endocrine insufficiency while 8 had pancreatic exocrine insufficiency preoperatively. All patients underwent pancreaticoduodenectomy. Observation indications: (1) surgical situations and follow-up; (2) analysis of influencing factors for pancreatic endocrine insufficiency after pancreaticoduodenectomy; (3) analysis of influencing factors for pancreatic exocrine insufficiency after pancreaticoduodenectomy. Follow-up using out-patient examination and telephone interview was performed to detect postoperative condition of blood glucose control, diet and nutrition, tumor recurrence and metastasis up to June 2018. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was analyzed using the independent sample t test. Measurement data with skewed distribution were described as M (range), and comparison between groups was analyzed using the Mann-Whitney U test. Count data were expressed as absolute numbers or percentages, and comparison between groups was analyzed using the chi-square test. Univariate analysis was conducted using the chi-square test. Multivariate analysis was conducted using the Logistic regression model. Results:(1) Surgical situations and follow-up: all the 168 patients underwent pancreaticoduodenectomy successfully and recovered well after operation. All patients were followed up for 6 months. The level of fasting and postprandial blood glucose of the 168 patients after surgery were 7 mmol/L(range, 5-9 mmol/L) and 10 mmol/L(range, 7-14 mmol/L), respectively. The defecation frequency was (2.4±1.2)times per day. No tumor recurrence or metastasis occurred in either patient. One hundred and thirty-two of the 168 patients were included in the study excepting patients with pancreatic endocrine insufficiency before operation. At postoperative 6 months, 47 patients developed pancreatic endocrine insufficiency, with an incidence of 35.61%(47/132). One hundred and sixty of the 168 patients were included in the study excepting patients with pancreatic exocrine insufficiency before operation. At postoperative 6 months, 68 patients had pancreatic exocrine insufficiency, with an incidence rate of 42.50%(68/160). (2) Analysis of influencing factors for pancreatic endocrine insufficiency after pancreaticoduodenectomy. Results of univariate analysis showed that gender, metabolic syndrome, chronic pancreatitis, excision point, and postoperative chemotherapy were the related factors for pancreatic endocrine insufficiency after pancreaticoduodenectomy ( χ2=5.300, 6.270, 4.473, 4.392, 5.397, P<0.05). Results of multivariate analysis revealed that male and metabolic syndrome were independent risk factors for pancreatic endocrine insufficiency after pancreaticoduodenectomy [ hazard ratio ( HR)=5.252, 5.364, 95% confidence interval ( CI): 1.362-6.382, 1.891-12.592, P<0.05)]. (3) Analysis of risk factors for pancreatic exocrine insufficiency after pancreaticoduodenectomy. Results of univariate analysis showed that body mass index (BMI), chronic pancreatitis, total bilirubin, excision point, postoperative pancreatic fistula as grade B or C, and pancreatic fibrosis were related factors for pancreatic exocrine insufficiency after pancreaticoduodenectomy ( χ2=1.691, 4.910, 7.763, 5.605, 4.663, 7.700, P<0.05). Results of multivariate analysis showed that BMI<18.5 kg/m 2, chronic pancreatitis, total bilirubin ≥171 μmol/L were independent risk factors for pancreatic exocrine insufficiency after pancreaticoduodenectomy ( HR=3.695, 5.231, 7.623, 95% CI: 1.232-7.324, 2.161-6.893, 1.562-5.235, P<0.05). Conclusions:Male and metabolic syndrome are risk factors for pancreatic endocrine insufficiency after pancreaticoduodenectomy. BMI<18.5 kg/m 2, chronic pancreatitis, and total bilirubin ≥171 μmol/L are risk factors for pancreatic exocrine insufficiency after pancreaticoduodenectomy.
		                        		
		                        		
		                        		
		                        	
5.Analysis of the clinical features of 20 cases of non-functioning pancreatic neuroendocrine tumors
Ge YAN ; Shanmei SHEN ; Shanhua BAO ; Yan BI ; Dalong ZHU
Chinese Journal of Postgraduates of Medicine 2019;42(2):127-130
		                        		
		                        			
		                        			Objective To investigate the clinical features of non- functioning pancreatic neuroendocrine tumors and to summarize experiences in the diagnosis and treatment. Methods The clinical pathology and follow-up data of 20 patients with non-functioning pancreatic neuroendocrine tumors treated in Nanjing Drum Tower Hospital Affiliated to Medical School of Nanjing University, from January 2008 to February 2018 were retrospectively analyzed. Results Among the 20 patients, 9 patients (9/20) were asymptomatic, and the tumor was found unexpectedly by physical examination. Eleven patients (11/20) with symptoms were admitted to the hospital due to non-specific gastrointestinal symptoms or compression symptoms caused by tumor space occupying. None of the patients had endocrine disorders. There were nineteen patients who received surgical treatment. According to the grading system, there were 7 patients (7/19) with G1, and 12 patients (12/19) with G2 . Among the 14 patients with diameter of tumor greater than 2 cm, there are 4 patients (4/14) with lymph node metastasis, and 4 patients (4/14) with liver metastasis. Five patients with diameter of tumor less than or equal to 2 cm had no distant metastasis. Conclusions NF-pNETs is often with nonspecific symptoms. Imaging examination is an important diagnostic method. Operation is the primary therapy for NF-pNETs. Because most of the small NF-pNETs with no symptoms are benign and grow slowly, and the metastatic rate is very low. It is important to weigh the pros and cons of surgical treatment for these patients.
		                        		
		                        		
		                        		
		                        	
6. Application of pegylated recombinant human granulocyte colony-stimulating factor to prevent chemotherapy-induced neutropenia in patients with lymphoma: a prospective, multicenter, open-label clinical trial
Huiqiang HUANG ; Bing BAI ; Yuhuan GAO ; Dehui ZOU ; Shanhua ZOU ; Huo TAN ; Yongping SONG ; Zhenyu LI ; Jie JIN ; Wei LI ; Hang SU ; Yuping GONG ; Meizuo ZHONG ; Yuerong SHUANG ; Jun ZHU ; Jinqiao ZHANG ; Zhen CAI ; Qingliang TENG ; Wanjun SUN ; Yu YANG ; Zhongjun XIA ; Hailin CHEN ; Luoming HUA ; Yangyi BAO ; Ning WU
Chinese Journal of Hematology 2017;38(10):825-830
		                        		
		                        			 Objective:
		                        			To evaluate the efficacy and safety of pegylated recombinant human granulocyte colony-stimulating factor (PEG-rhG-CSF) in prophylaxis neutropenia after chemotherapy in patients with lymphoma.
		                        		
		                        			Methods:
		                        			This was a multicenter, single arm, open, phase Ⅳ clinical trial. Included 410 patients with lymphoma received multiple cycles of chemotherapy and PEG-rhG-CSF was administrated as prophylactic. The primary endpoint was the incidence of Ⅲ/Ⅳ grade neutropenia and febrile neutropenia (FN) after each chemotherapy cycle. Meanwhile the rate of antibiotics application during the whole period of chemotherapy was observed.
		                        		
		                        			Results:
		                        			①Among the 410 patients, 8 cases (1.95%) were contrary to the selected criteria, 35 cases (8.54%) lost, 19 cases (4.63%) experienced adverse events, 12 cases (2.93%) were eligible for the termination criteria, 15 cases (3.66%) develpoed disease progression or recurrence, thus the rest 321 cases (78.29%) were into the Per Protocol Set. ②During the first to fourth treatment cycles, the incidences of grade Ⅳ neutropenia after prophylactic use of PEG-rhG-CSF were 19.14% (49/256) , 12.5% (32/256) , 12.18% (24/197) , 13.61% (20/147) , respectively. The incidences of FN were 3.52% (9/256) , 0.39% (1/256) , 2.54% (5/197) , 2.04% (3/147) , respectively. After secondary prophylactic use of PEG-rhG-CSF, the incidences of Ⅳ grade neutropenia decreased from 61.54% (40/65) in the screening cycle to 16.92% (11/65) , 18.46% (12/65) and 20.75% (11/53) in 1-3 cycles, respectively. The incidences of FN decreased from 16.92% (11/65) in the screening cycle to 1.54% (1/65) , 4.62% (3/65) , 3.77% (2/53) in 1-3 cycles, respectively. ③The proportion of patients who received antibiotic therapy during the whole period of chemotherapy was 34.39% (141/410) . ④The incidence of adverse events associated with PEG-rhG-CSF was 4.63% (19/410) . The most common adverse events were bone pain[3.90% (16/410) ], fatigue (0.49%) and fever (0.24%) .
		                        		
		                        			Conclusion
		                        			During the chemotherapy in patients with lymphoma, the prophylactic use of PEG-rhG-CSF could effectively reduce the incidences of grade Ⅲ/Ⅳ neutropenia and FN, which ensures that patients with lymphoma receive standard-dose chemotherapy to improve its cure rate. 
		                        		
		                        		
		                        		
		                        	
7.Analysis of prognostic factors of portal hypertension treated with devascularization
Yajuan CAO ; Yiming PAN ; Shanhua BAO ; Chenglin LU ; Biyun XU ; Min XIE
Chinese Journal of Surgery 2016;54(6):434-438
		                        		
		                        			
		                        			Objective To explore the prognostic factors of portal hypertension treated with devascularization.Methods A total of 397 patients with portal hypertension underwent devascularization in Nanjing Drum Tower Hospital from February 1993 to April 2014,among which there were 242 male and155 female patients with median age of 48 years.The perioperative data were retrospectively collected.Logistic regression was used to find the risk factors which affect the operative complications.Follow-up evaluation was in progress regularly.Kaplan-Meier survival curve,Log-rank test and Cox regression model were used to find out factors which affect the long-term results.Results All together 397 patients underwent devascularization,in whom 8 patients died perioperative,389 patients discharged successfully.Logistic regression showed that age (≥ 48 years) (X2 =4.559,OR =2.048,P =0.033),red color sign before surgery (x2 =4.959,OR =2.129,P =0.026) and without portosystemic collateral vessels reserved (x2 =13.348,OR =5.122,P =0.000) were risk factors of perioperative complications.The follow-up time was (5.7±4.6) years.Totally 27 patients were lost from follow-up,103 patients died for the disease during follow-up.The survival rate at 1-,3-,5-,10-,15-and 20-years was 93.6%,86.9%,80.1%,59.3%,54.1% and 38.5% respectively.Univariate analysis showed that gender (male),age (≥ 48 years),hemorrhage before surgery (≥500 ml per time),hepatitis virus and without portosystemic collateral vessels reserved were risk factors of the long-term survival (P < 0.05).Cox regression analysis showed that age (≥48 years) (X2 =9.850,RR =1.904,P =0.002),hemorrhage before surgery (≥ 500 ml per time)(X2 =34.402,RR =3.273,P =0.000),hepatitis virus (X2 =7.573,RR =2.525,P =0.006) and without portosystemic collateral vessels reserved (x2 =5.905,RR =1.889,P =0.015) were independent risk factors that affect the long-term survival.Conclusion Devascularization with portosystemic collateral vessels reserved has favorable perioperative and long-term outcome,and it definitely is a very safe and effective technique for portal hypertension.
		                        		
		                        		
		                        		
		                        	
8.Analysis of prognostic factors of portal hypertension treated with devascularization
Yajuan CAO ; Yiming PAN ; Shanhua BAO ; Chenglin LU ; Biyun XU ; Min XIE
Chinese Journal of Surgery 2016;54(6):434-438
		                        		
		                        			
		                        			Objective To explore the prognostic factors of portal hypertension treated with devascularization.Methods A total of 397 patients with portal hypertension underwent devascularization in Nanjing Drum Tower Hospital from February 1993 to April 2014,among which there were 242 male and155 female patients with median age of 48 years.The perioperative data were retrospectively collected.Logistic regression was used to find the risk factors which affect the operative complications.Follow-up evaluation was in progress regularly.Kaplan-Meier survival curve,Log-rank test and Cox regression model were used to find out factors which affect the long-term results.Results All together 397 patients underwent devascularization,in whom 8 patients died perioperative,389 patients discharged successfully.Logistic regression showed that age (≥ 48 years) (X2 =4.559,OR =2.048,P =0.033),red color sign before surgery (x2 =4.959,OR =2.129,P =0.026) and without portosystemic collateral vessels reserved (x2 =13.348,OR =5.122,P =0.000) were risk factors of perioperative complications.The follow-up time was (5.7±4.6) years.Totally 27 patients were lost from follow-up,103 patients died for the disease during follow-up.The survival rate at 1-,3-,5-,10-,15-and 20-years was 93.6%,86.9%,80.1%,59.3%,54.1% and 38.5% respectively.Univariate analysis showed that gender (male),age (≥ 48 years),hemorrhage before surgery (≥500 ml per time),hepatitis virus and without portosystemic collateral vessels reserved were risk factors of the long-term survival (P < 0.05).Cox regression analysis showed that age (≥48 years) (X2 =9.850,RR =1.904,P =0.002),hemorrhage before surgery (≥ 500 ml per time)(X2 =34.402,RR =3.273,P =0.000),hepatitis virus (X2 =7.573,RR =2.525,P =0.006) and without portosystemic collateral vessels reserved (x2 =5.905,RR =1.889,P =0.015) were independent risk factors that affect the long-term survival.Conclusion Devascularization with portosystemic collateral vessels reserved has favorable perioperative and long-term outcome,and it definitely is a very safe and effective technique for portal hypertension.
		                        		
		                        		
		                        		
		                        	
9.Intraoperation localization of occult insulinoma by using step by step occlusion of the pancrease
Jian WANG ; Hui ZHANG ; Min XIE ; Shanhua BAO
Chinese Journal of Endocrine Surgery 2015;(6):473-475,479
		                        		
		                        			
		                        			Objective To explore clinical efficacy and safety of intraoperative localization of occult insu -linoma by using step-by-step occlusion of the pancreas .Methods 22 cases of occult insulinoma patients admit-ted from Mar.2003 to May 2013 were given intraoperative localization by adopting the technology of step -by-step occlusion of the pancreas .Results All the 22 patients were successfully completed the segmental resection of pancreas.The average operation time was(120 ±50)min, and the average intraoperative blood loss was (100 ± 80)ml.No blood transfusion was needed.Blood glucose rose rapidly when insulinoma was located within the scope of occlusion, blood glucose remained unchanged when insulinoma was beyond the scope of occlusion ,and blood glucose dropped swiftly when insulinoma was on the point of occlusion .Two patients had postoperative short-term pancreatic fistula and they were cured by conservative treatment .No other complications occurred .The average hospitalization time was(12 ±5)d.The result was good during the followed up of 8 to 24 months.Con-clusion The technique of step-by-step occlusion of the pancreas for localization of occult insulinoma is effective supplement for conventional methods , worthy of promotion .
		                        		
		                        		
		                        		
		                        	
10.Preoperative T staging of gastric cancer: comparison between MR including diffusion weighted imaging and contrast enhanced CT scan.
Song LIU ; Jian HE ; Wenxian GUAN ; Qiang LI ; Zhuping ZHOU ; Haiping YU ; Shanhua BAO ; Zhengyang ZHOU
Chinese Journal of Gastrointestinal Surgery 2014;17(3):245-249
OBJECTIVETo compare the accuracy of CT and MR including diffusion-weighted imaging(DWI) in preoperative diagnosis and T staging of gastric cancer.
METHODSForty-one patients with gastric cancers proved by gastroscopy biopsy from November 2011 to August 2012 were prospectively enrolled. They underwent contrast enhanced CT and MR imaging (including DWI, T2 weighted and dynamic enhanced imaging) preoperatively. Two radiologists interpreted CT and MR images for detecting and staging each patient independently. With the reference of post-operative histopathological findings, T staging accuracy of CT and MR imaging was calculated and compared. Inter-observer agreement was also evaluated.
RESULTSOverall T staging accuracy in MR including DWI was significantly higher than that in CT imaging(87.8% vs. 65.9%, P=0.004). MR had a better inter-observer agreement than CT(Kappa=0.813, 0.603, respectively).
CONCLUSIONMR including DWI can improve preoperative T staging accuracy of gastric cancer significantly, which deserves recommendation for clinical application.
Biopsy ; Contrast Media ; Diffusion Magnetic Resonance Imaging ; Gastroscopy ; Humans ; Magnetic Resonance Imaging ; Neoplasm Staging ; Stomach Neoplasms ; pathology
            
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