Objective:To investigate the effect of β-Ecdysterone (β-Ecd), an active ingredient in cow′s knee, on hormonal femoral head necrosis in young rabbits and explore the mechanism involved.Methods:An animal study.Sixty New Zealand young rabbits were divided into control, model and intervention groups by random number table method, with 20 rabbits in each group.Prednisolone acetate (7.5 mg/kg) was injected bilaterally into the gluteal muscle of rabbits in both model and intervention groups twice a week.β-Ecd (0.5 mg/kg) was injected subcutaneously at the time of the first injection of Prednisolone acetate in the intervention group for 5 times a week.An equal amount of saline was injected into rabbits in control and model groups.Eight weeks after modelling, animals were put to death, and femoral heads were taken from both sides for gross observation.Micro computed tomography(Micro-CT) was used to analyze the microstructure of bone trabeculae and to measure bone microstructural parameters.Histological staining was used to detect changes in the morphology of bone tissues.Immunohistochemistry and Western blot were used to examine the expression of osteogenic and lipogenic factors and proteins in the femoral head tissue.Rabbit bone marrow mesenchymal stem cells (BMSCs) were divided into a blank control group, a Dexamethasone group and a β-Ecd intervention group.Cell damage was induced by Dexamethasone in Dexamethasone group and β-Ecd intervention group, and the β-Ecd intervention group was given the optimal concentration of β-Ecd.Western blot was used to detect the expression of osteogenic and lipogenic proteins in the cells of each group.Real-time quantitative polymerase chain reaction (RT-qPCR) was used to determine the mRNA expression of osteogenic and lipogenic marker genes.Results:After excluding 9 rabbits that died during the experimental period, 51 rabbits were finally included in the study, with 19 in the control group, 15 in the model group and 17 in the intervention group.Gross observation and Micro-CT showed that compared with that of the control group, the femoral head of the model group was obscure and greyish, with dark red necrotic areas.The bone trabeculae of the model group were sparse, thinned, disordered, and partially fractured, compared with those of the control group.The changes in the femoral head and bone trabeculae of the intervention group were between those of control and model groups.The bone mineral density, the number, thickness and relative volume of bone trabeculae significantly decreased and trabecular separation significantly increased in both model and intervention groups, compared with those in the control group ( F=12.78, 45.52, 32.74, 64.08, 8.83, all P<0.05).However, these symptoms in the intervention group were better than those in the model group.Pathological histology showed that in the control group, bone trabeculae were neatly arranged, robust and full, with a high number of osteoclasts and occasional empty bone sockets.In the model group, bone trabeculae were sparsely arranged and broken, with fewer osteoclasts, and the number of empty bone sockets increased and enlarged.In the intervention group, bone trabeculae had a more complete morphology, with fewer necrotic osteoclasts and reduced empty bone sockets, compared with the model group.Immunohistochemistry results showed that compared with the control group, the model group and intervention group had increased content of fatty acid binding proteins (FABP) and CCAAT/enhancer binding proteins α (CEBP) in the femoral head bone tissue, and decreased content of osteopontin (OPN) and Runt-related transcription factor 2 (RUNX2) .The changes in each index were greater in the model group than those in the intervention group ( F=21.07, 24.06, 17.92, 21.36, all P<0.05). Western blot detection showed that compared with the control group, the CEBP protein expression content of the femoral head in the model group and the intervention group was increased and the RUNX2 protein expression content was decreased. The changes of CEBP and RUNX2 were greater in the model group than those in the intervention group( F=73.43, 197.87, all P<0.05).Western blot detection of BMSCs showed that compared with the blank control group, the Dexamethasone group and β-Ecd intervention group had decreased expression of OPN and RUNX2 proteins and increased expression of FABP and CEBP proteins ( F=161.61, 358.01, 91.18, 69.04, all P<0.05).The changes in each index in the β-Ecd intervention group were smaller than those in the Dexamethasone group.RT-qPCR detection of BMSCs showed that in the Dexamethasone group had lower expression of OPN and RUNX2 and higher expression of CEBP and FABP than the blank control group ( F=19.71, 45.08, 61.46, 15.12, all P<0.05).The changes in each index in the β-Ecd intervention group were smaller than those in the Dexamethasone group. Conclusions:β-Ecd can attenuate the pathological changes of hormonal femoral head necrosis in young rabbits by promoting osteogenic differentiation of BMSCs, inhibiting lipogenic differentiation of BMSCs, and improving bone microstructure.

Objective:To investigate the effect of β-Ecdysterone (β-Ecd), an active ingredient in cow′s knee, on hormonal femoral head necrosis in young rabbits and explore the mechanism involved.Methods:An animal study.Sixty New Zealand young rabbits were divided into control, model and intervention groups by random number table method, with 20 rabbits in each group.Prednisolone acetate (7.5 mg/kg) was injected bilaterally into the gluteal muscle of rabbits in both model and intervention groups twice a week.β-Ecd (0.5 mg/kg) was injected subcutaneously at the time of the first injection of Prednisolone acetate in the intervention group for 5 times a week.An equal amount of saline was injected into rabbits in control and model groups.Eight weeks after modelling, animals were put to death, and femoral heads were taken from both sides for gross observation.Micro computed tomography(Micro-CT) was used to analyze the microstructure of bone trabeculae and to measure bone microstructural parameters.Histological staining was used to detect changes in the morphology of bone tissues.Immunohistochemistry and Western blot were used to examine the expression of osteogenic and lipogenic factors and proteins in the femoral head tissue.Rabbit bone marrow mesenchymal stem cells (BMSCs) were divided into a blank control group, a Dexamethasone group and a β-Ecd intervention group.Cell damage was induced by Dexamethasone in Dexamethasone group and β-Ecd intervention group, and the β-Ecd intervention group was given the optimal concentration of β-Ecd.Western blot was used to detect the expression of osteogenic and lipogenic proteins in the cells of each group.Real-time quantitative polymerase chain reaction (RT-qPCR) was used to determine the mRNA expression of osteogenic and lipogenic marker genes.Results:After excluding 9 rabbits that died during the experimental period, 51 rabbits were finally included in the study, with 19 in the control group, 15 in the model group and 17 in the intervention group.Gross observation and Micro-CT showed that compared with that of the control group, the femoral head of the model group was obscure and greyish, with dark red necrotic areas.The bone trabeculae of the model group were sparse, thinned, disordered, and partially fractured, compared with those of the control group.The changes in the femoral head and bone trabeculae of the intervention group were between those of control and model groups.The bone mineral density, the number, thickness and relative volume of bone trabeculae significantly decreased and trabecular separation significantly increased in both model and intervention groups, compared with those in the control group ( F=12.78, 45.52, 32.74, 64.08, 8.83, all P<0.05).However, these symptoms in the intervention group were better than those in the model group.Pathological histology showed that in the control group, bone trabeculae were neatly arranged, robust and full, with a high number of osteoclasts and occasional empty bone sockets.In the model group, bone trabeculae were sparsely arranged and broken, with fewer osteoclasts, and the number of empty bone sockets increased and enlarged.In the intervention group, bone trabeculae had a more complete morphology, with fewer necrotic osteoclasts and reduced empty bone sockets, compared with the model group.Immunohistochemistry results showed that compared with the control group, the model group and intervention group had increased content of fatty acid binding proteins (FABP) and CCAAT/enhancer binding proteins α (CEBP) in the femoral head bone tissue, and decreased content of osteopontin (OPN) and Runt-related transcription factor 2 (RUNX2) .The changes in each index were greater in the model group than those in the intervention group ( F=21.07, 24.06, 17.92, 21.36, all P<0.05). Western blot detection showed that compared with the control group, the CEBP protein expression content of the femoral head in the model group and the intervention group was increased and the RUNX2 protein expression content was decreased. The changes of CEBP and RUNX2 were greater in the model group than those in the intervention group( F=73.43, 197.87, all P<0.05).Western blot detection of BMSCs showed that compared with the blank control group, the Dexamethasone group and β-Ecd intervention group had decreased expression of OPN and RUNX2 proteins and increased expression of FABP and CEBP proteins ( F=161.61, 358.01, 91.18, 69.04, all P<0.05).The changes in each index in the β-Ecd intervention group were smaller than those in the Dexamethasone group.RT-qPCR detection of BMSCs showed that in the Dexamethasone group had lower expression of OPN and RUNX2 and higher expression of CEBP and FABP than the blank control group ( F=19.71, 45.08, 61.46, 15.12, all P<0.05).The changes in each index in the β-Ecd intervention group were smaller than those in the Dexamethasone group. Conclusions:β-Ecd can attenuate the pathological changes of hormonal femoral head necrosis in young rabbits by promoting osteogenic differentiation of BMSCs, inhibiting lipogenic differentiation of BMSCs, and improving bone microstructure.

Objective:To analyze the mutations of globin genes among patients suspected for thalassemia from the Shanghai area.Methods:A total of 4 644 patients diagnosed at Ruijin Hospital, Shanghai Jiao Tong University School of Medicine between June 2016 and December 2019 were selected as the study subjects. The patients were tested for common mutations associated with thalassemia gene by Gap-PCR and reverse dot blotting (RDB). Patients were suspected to harbor rare mutations based on the inconsistency between hematological phenotypes and results of common mutation detection, and were further analyzed by Gap-PCR and Sanger sequencing.Results:Among the 4 644 patients, 2 194 (47.24%) were found to carry common thalassemia mutations, among which 701 (15.09%) were α-thalassemia, 1 448 (31.18%) were β-thalassemia, and 45 (0.97%) were both α- and β-thalassemia. Forty six samples were found to harbor rare mutations, which included 17 α-globin gene and 29 β-globin gene mutations. CD77(CCC>ACC) ( HBA2: c. 232C>A) of the α-globin gene, NG_000007.3: g. 70567_71015del449, codon 102(-A) ( HBB: c. 308_308delA) and IVS-Ⅱ-636 (A>G) ( HBB: c. 316-215A>G) of the β-globin gene were previously unreported new types of globin gene mutations. Conclusion:Among the 4 644 patients, the detection rate for common thalassemia mutations was 47.24%, whilst 46 samples were detected with rare gene mutations. The type of gene mutation types were diverse in the Shanghai area. The study has provided more accurate results for genetic diagnosis and counseling.

【Objective】 To develop and verify a kinetic method for the determination of prekallikrein activator (PKA) content. 【Methods】 The optimal reaction conditions were determined by comparing the factors of pH and ionic strength of different sample dilution buffers, incubation time of each procedure, and incubation temperature. The accuracy, specificity, precision, linearity, stability and durability of the method were validated. 【Results】 The sample was diluted with 0.05 mol/L Tris-HCl buffer (pH8.5, containing 0.15 mol/L NaCl) and incubated by prekallikrein (PK) at 37℃ for 20 min. After that, the substrate S-2302 was added. Within 10 min before the measurement, the absorbance change rate reached △A405/min. The validation results indicated that the linear range of the method was (0.5~4.0)IU/mL, while the recovery of calibration standard was 96.9%~103.7% with the R² value more than 0.99. The specificity test showed that human serum albumin, excipients of intravenous human immunoglobulin (pH4), low pH and protein content had no significant effect on the detection of PKA, The recovery rates of standard sample solution in the specificity experiment were 98.0% (0.9% sodium chloride solution), 95.3% (0.46% sodium caprylate solution), 96.7% (10% maltose solution, pH4.0), 94.0%(20%BSA), and 94.0%(5%BSA, pH4.0), respectively. The accuracy and precision of the method can meet requirements in the range between 0.5 and 4.0 IU/mL. The inter-batch recovery rate of quality control samples were between 96.4%~109.5% with the coefficients of variation(CV) between 0.2%~6.9%, while the intra-batch recovery rate were between 101.5%~102.9% with the CV between 2.6%~5.9%. The linearity, accuracy and precision of the assay can meet the requirements when PK and S-2302 were placed at room temperature for less than 6 hours, with the recovery rate of quality control samples between 94.9%~109.9%. The end-point method and kinetic method were used to determine the PKA in 20 batches of human serum albumin, and the consistency showed that there was no significant difference between the two methods(P>0.05). 【Conclusion】 A kinetic method for determination of PKA content with good linearity, specificity, accuracy, precision, stability and durability has been established. Compared with the method in ChP, the new method is more convenient, accurate and rapid to determine the content of PKA in human albumin and human immunoglobulin (pH4) for intravenous injection.

Objective: To summary the experience of 233 cases of laparoscopic pancreaticoduodenectomy (LPD) performed by a single surgical team. Methods: Data of patients undergoing LPD from September 2012 to October 2016 were reviewed. There were 145 males and 88 females with the mean age of(60.3±13.0)years old, ranging from 19 to 92 years old, and the mean body mass index of (22.8±3.5)kg/m^2, ranging from 16.3 to 36.8 kg/m². There were 195 patients with clinical manifestation and 54 patients who had the history of abdominal surgery. Results: LPD were performed on 233 patients by same surgical team consecutively. The mean operative time was(368.0±57.4)minutes. Mean blood loss was(203.8±138.6)ml. The postoperative morbidity rate was 33.5%, with 6.9% of grade B or C pancreatic fistula and 9.9% of bleeding. The reoperation rate was 5.6%. The mortality during 30 days after operation was 0.9%. Mean postoperative hospital stay was (18.1±11.2)days. Mean tumor size was (3.9±2.4)cm, and the mean number of lymph nodes harvested was 21.3±11.9.One hundred and sixty-three patients were diagnosed as malignant tumor, including pancreatic adenocarcinoma(n=84), cholangiocarcinoma(n=17), ampullary adenocarcinoma(n=55), duodenal adenocarcinoma(n=5), gastric cancer(n=1)and duel cancer (n=1) located in distal stomach and duodenum. Conclusion The key point to make laparoscopic pancreaticduodenectomy a routine safe procedure is to operate the procedure under skilled hands in selected patients via suitable surgical approaches.

Objective To evaluate the CT and MRI imaging features of primary ventricular lymphoma ,so as to provide a reference for the diagnosis and differential diagnosis of the disease .Methods The clinical data and imaging findings of primary ventricular lymphoma in one case was retrospectively analyzed with literature review and compared with primary lymphoma in parenchymal brain .Results Slightly high density tumor presented with regular shape in the trigone of lateral ventricles on CT findings , and slightly low signal intensity on T 1 and slightly high signal intensity on T 2 appeared on MRI findings with small cystic necrosis in tumour .An enhancement scan showed obviously enriched, and there was perilesional edema.After 14 day’s dehydration treatment, rechecked indicated that the lesions were significantly en-larged, the shape were irregula , the area of cystic necrosis was enlarged , heterogeneous enhanced scanning were enhanced , perilesional ede-ma became more heavier , and the imaging feature of “closed fist” appeared .It was proved to be diffuse large B cell lymphoma with surgical pathologic examination , and there was no lymphoma in other regions of the body with whole body PET-CT examination , which was clinically diagnosed as primary ventricular lymphoma .Conclusion Primary ventricular lymphoma is rarely reported ,and it has similar imaging mani-festations with primary lymphoma in parenchymal brain .The combination of multiple imaging modalities will be helpful to diagnosis and dif -ferential diagnosis for the suspected patients .

Objective To investigate the difference in the vulnerability of carotid atherosclerotic plaques in patients with unilateral and bilateral intraplaque hemorrhage (IPH). Methods A retrospective analysis was conducted among 44 patients with unilateral IPH (30 cases) or bilateral IPH (14 cases) in the carotid plaques detected by magnetic resonance imaging (MRI) in our hospital between December, 2009 and December, 2012. The age, maximum wall thickness and incidence of fibrous cap rupture were compared between the two groups. Results Compared with those with unilateral IPH, the patients with bilateral IPHs had a significantly younger age (66.6 ± 9.4 years vs 73.7 ± 9.0 years, P=0.027), a significantly greater maximum plaque thickness (6.3 ± 1.9 mm vs 5.0 ± 1.3 mm, P=0.035) and a higher incidence of ulcers (50% vs 13.3%, P=0.025). Logistic regression analysis revealed a significant association between bilateral IPHs and the occurrence of ulcer with an odd ratio (OR) of 6.5 (95%confidence interval [CI]:1.5-28.7, P=0.014). After adjustment for gender in Model 1, bilateral IPHs were still significantly associated with presence of ulcer (OR=5.7, 95%CI: 1.1-29.2, P=0.036). But after adjustment for age (P=0.131) or maximum plaque thickness (P=0.139) in model 2, no significant correlation was found between bilateral IPHs and the presence of ulcer. Conclusion Compared with patients with unilateral IPH, those with bilateral IPHs are at a younger age and have a greater plaque burden and a higher incidence of fibrous cap rupture, suggesting a greater vulnerability of the carotid plaques in patients with bilateral IPHs.

Objective To investigate the difference in the vulnerability of carotid atherosclerotic plaques in patients with unilateral and bilateral intraplaque hemorrhage (IPH). Methods A retrospective analysis was conducted among 44 patients with unilateral IPH (30 cases) or bilateral IPH (14 cases) in the carotid plaques detected by magnetic resonance imaging (MRI) in our hospital between December, 2009 and December, 2012. The age, maximum wall thickness and incidence of fibrous cap rupture were compared between the two groups. Results Compared with those with unilateral IPH, the patients with bilateral IPHs had a significantly younger age (66.6 ± 9.4 years vs 73.7 ± 9.0 years, P=0.027), a significantly greater maximum plaque thickness (6.3 ± 1.9 mm vs 5.0 ± 1.3 mm, P=0.035) and a higher incidence of ulcers (50% vs 13.3%, P=0.025). Logistic regression analysis revealed a significant association between bilateral IPHs and the occurrence of ulcer with an odd ratio (OR) of 6.5 (95%confidence interval [CI]:1.5-28.7, P=0.014). After adjustment for gender in Model 1, bilateral IPHs were still significantly associated with presence of ulcer (OR=5.7, 95%CI: 1.1-29.2, P=0.036). But after adjustment for age (P=0.131) or maximum plaque thickness (P=0.139) in model 2, no significant correlation was found between bilateral IPHs and the presence of ulcer. Conclusion Compared with patients with unilateral IPH, those with bilateral IPHs are at a younger age and have a greater plaque burden and a higher incidence of fibrous cap rupture, suggesting a greater vulnerability of the carotid plaques in patients with bilateral IPHs.

Objective To improve the diagnostic accuracy by analyzing the computed tomography ( CT) characteristic performance of mucoepidermoid carcinoma of the lung. Methods CT findings 15 cases proved by pathology of mucoepidermoid carcinoma of the lung were retrospectively analyzed,and CT characteristic performance were studied by reviewing the literatures. Results The mucoepidermoid carcino-ma were located in right bronchial of 8 cases,left bronchial of 6 cases,peripheral part of the lung of 1 case. Calcification was seen in 7 mas-ses. Most masses density was not homogeneous at plain scanning. Venous phase have significantly enhanced compared with arterial phase. Ten cases showed obstructive pneumonia and atelectasis,3 cases with mediastinal lymph node metastases. Conclusion Multiple mucoepidermoid carcinoma of the lung grows extension of the long axis of bronchial,calcification be seen in most of patients. The enhanced mode of the lesions appeared as continuous progressive enhancement,some were illustrated with post obstructive pneumonia or atelectasis,CT have a certain value of diagnosis and diffenertial diagnosis.

Objective To investigate clinical efficacy of Easy First strategy in laparoscopic pancreaticoduodenectomy (PD) for borderline resectable pancreatic cancer.Methods The clinical data of 9 patients with borderline resectable pancreatic cancer who were admitted to the Sir Run Run Shaw Hospital of Zhejiang University (8 patients) and Zhejiang Provincial People's Hospital (1 patient) from June 2013 to March 2015 were retrospectively analyzed.Nine patients underwent laparoscopic pancreaticoduodenectomy based on the Easy First strategy (to sequentially dissect and amputate jejunum,stomach,hepatoduodenal ligament,common bile duct,main portal vein,head of pancreas,second segment and third segment of the duodenum,uncinate process and neck of pancreas).The operation time,volume of intraoperative blood loss,postoperative pathological examination,complications and duration of hospital stay were recorded.Patients were followed up once every 3 months by outpatient examination and telephone interview up to June 2015.Results Of 9 patients,4 received totally laparoscopic PD (2 received partial resection and repair of portal venous wall),1 received laparoscopic assisted resection and digestive tract reconstruction,and 4 received laparoscopic transection of jejunum,bile tract and stomach and conversion to open surgery for resection and digestive tract reconstruction (1 with severe adhesion between tumor and portal vein,3 with bleeding due to dissection of neck and unciform process of pancreas).The operation time and volume of intraoperative blood loss in all patients and in 4 patients with totally laparoscopic PD were (404 ± 49) minutes and (456 ± 348) mL,(395 ± 61) minutes and (188 ± 25) mL,respectively.Of 9 patients,5 with postoperative complications were cured without perioperative death,including 2 with Grade B pancreatic fistula,1 with biliary leakage,1 receiving reoperation due to gastric stump bleeding at postoperative day 7 and 1 with abdominal infection.The extubation time of right drainage tube and left drainage tube was (9 ± 5) days and (11 ± 4) days,respectively.The duration of hospital stay was (24 ± 10)days.All patients were diagnosed as with pancreatic cancer by pathological examinations with the tumor diameter of (3.2 ± 0.8) cm.The number of harvested lymph nodes in all patients and in 4 patients with totally laparoscopic PD were 16.8 (range,6.0-25.0) and 19.8 (range,15.0-25.0).All the patients were followed up for mean time of 12 months (range,4-24 months),including 1 death at postoperative month 3,1 with tumor survival of 20 months and others with tumor-free survival.The postoperative survival time of 4 patients was more than 18 months at the end of follow-up.Conclusion Easy First strategy in laparoscopic PD is safe,feasible and practical for borderline resectable pancreatic cancer.