Objective To study the chemical constituents of Paeonia veitchii. Methods Isolation and purification were carried out on repeated silica gel column chromatography. The structures of the compounds were identified by physico-chemical properties and spectral analyses. Results Nine compounds were isolated and identified as paeoniflorin (Ⅰ), hydroxypaeoniflorin (Ⅱ), benzoylpaeoniflorin (Ⅲ), benzoylhydroxypaeoniflorin (Ⅳ), albiflorin (Ⅴ), paeonisothujone (Ⅵ), mudanpinoic acid A (Ⅶ), 2-hydroxybenzyl alcohol (Ⅷ), bis (2-hydroxybenzyl) ether (Ⅸ). Conclusion Compound Ⅸ is obtained from natural products for the first time. Compounds Ⅳ, Ⅵ-Ⅸ are isolated from this plant for the first time.

Objective To establish a HRM assay to screen for KRAS mutations in clinical colorectal cancer patients.Methods The sensitivity of HRM was analyzed by detecting somatic mutations in exon 2,notably codons 12 and 13 of the KRAS gene in the serial plasmid mixture samples which were mixed using the different proportions mutation plasmid and wide type plasmid of KRAS.HRM analysis was performed for KRAS on DNA insolated from a panel of 60 colorectal cancer samples derived from fresh tissues.The results were compared with the direct sequencing data.Results After the PCR amplification,the mutation results could be available by performing HRM analysis in the same tube on a real time PCR machine with HRM capability.HRM detection could identify KRAS mutation in a proportion of 10% of mutation plasmid DNA.All 60 samples identified the KRAS mutation by HRM and sequencing.17 samples were positive(28.3%) by HRM for KRAS exon 2 mutations,and 15 samples were confirmed the presence of codon 12 or 13 mutations(25.0%) and the other 2 samples were wild type by sequencing.The 60 samples detected by HRM were given 100% sensitivity with 96% specificity.Conclusions HRM is a sensitive intube methodology to screen for mutations in clinical samples.HRM will enable high-throughput screening to gene mutations to allow appropriate therapeutic choices for patients and accelerate research aimed at identifying novel mutations in human cancer.

Objective To investigate the risk factors for clinical poor outcome after microsurgical treatment of ruptured middle cerebral artery aneurysms (MCAA).Methods The patients with ruptured MCAA treated with microsurgery were enrolled retrospectively.The data of demography,medical history,Hunt-Hess grade,imaging characteristics,surgery-related factors,and postoperative complications were collected.Glasgow Outcome Scale (GOS) was used to assess the outcomes.GOS 4-5 was defined as a good outcome and 1-3 was defined as a poor outcome.Results A total of 44 patients with ruptured MCAA were enrolled,including 26 (59.1％) females and 18 (40.9％) males,aged 36-75 years old (mean 58.5 years).The aneurysms were clipped in 42 cases (95.5％) and wrapped in 2 eases (4.5％).They were followed up for 3-6 months,33 (75.0％) had good outcome and 11 (25.0％) had poor outcome.Univariate analysis showed that there were significant differences in the proportions of Hunt-Hess grade,midline shift degree,hematoma volume,intraoperative rupture,decompressive craniectomy,and postoperative pulmonary infection between the good outcome group and the poor outcome group.Multivariatelogistic regression analysis showed that Hunt-Hess grade Ⅳ-Ⅴ (odds ratio [OR] 20.885,95％ confilence interval [CI] 1.342-38.696;P =0.001),intraoperative anenrysm rupture (OR 18.906,95％ CI 2.918-20.915;P=0.011),and complicated with pulmonary infection (OR 38.865,95％ CI 18.718-40.509;P =0.001) were the independent risk factors for poor outcomes.Conclusion The high Hunt-Hess grade,intraoperative aneurysm rupture,and complicated with pulmonary infection after surgery are the independent risk factors for poor outcomes after microsurgical treatment in patients with ruptured MCAA.

Objective To compare the effect of the bipolar electric coagulation and unipolar electric coagulation on cranioplasty of scalp separation. Methods The clinical data of 67 patients who underwent unilateral frontotemporal cranioplasty from 2014 to 2017 were retrospectively analyzed. According to coagulation method during operation, these patients were divided into two groups, unipolar electric coagulation group (32 cases) and bipolar electric coagulation group (35 cases). The operation time, postoperative intracranial hemorrhage, infection, epilepsy and subcutaneous effusion were compared between two groups. Results The operation time of two groups had no significant difference (P > 0.05). The incidence of intracranial hemorrhage, infection and epilepsy of two groups had no significant differences (P > 0.05). But the incidence of subcutaneous effusion in unipolar electric coagulation group was significantly higher than that in bipolar electric coagulation group: 28.1%(9/32) vs. 5.7%(2/35), P<0.05. Conclusions The use of unipolar electric coagulation during the scalp separation in cranioplasty can reduce operation time in a certain extent, but significantly increase the incidence of postoperative subcutaneous effusion.

OBJECTIVETo identify the endophytic fungal strain PR35 separated from Paeonia delavayi and study chemical constituents of its secondary metabolites.

METHODThe fungal strain PR35 was identified by morphological observation and ITS rDNA sequence analysis. Various chromatographic methods were adopted to separate and purify its secondary metabolites, and their structures were identified by physiochemical properties and spectral data

RESULTThe fungal strain PR35 was identified as Trichoderma longibrachiatum. Five compounds were separated from fermentation products of fungal strain PR35 and identified as 1-(2,6-dihydroxyphenyl)-3-hydroxybutan-1-one (1), 1-(2,6-dihydroxypheny) propan-1-one (2), 1-(2,4,6-trihydroxyphenyl) butan-1-one (3), 4-methoxy-1-naphthol (4), and cerevisterol (5). Among them, compounds 1-3 showed notable antifungal activities against Botrytis cinerea, Fusarium avenaceum and Hormodendrum compactum.

CONCLUSIONThe endophytic fungus T. longibrachiatum was separated from the plant P. delavayi for the first time. Five compounds were first separated from endophytic fungus of P. delavayi. Among them, compound 4 was separated from microbial fermentation products for the first time.

DNA, Fungal ; genetics ; DNA, Intergenic ; genetics ; Endophytes ; classification ; genetics ; isolation & purification ; metabolism ; Paeonia ; microbiology ; Phylogeny ; Trichoderma ; classification ; genetics ; isolation & purification ; metabolism

Objective To investigate the efficacy of neuroendoscopy combined with urokinase in the treatment of spontaneous intraventricular hemorrhage.Methods From August 2014 to August 2017,91 spontaneous thalamic hemorrhage ruptured into ventricles patients in Affiliated Hospital of the Yangzhou University were enrolled,who were underwent surgical treatment in this retrospective study.The patients were divided into the study group(n =41) and control group(n =50) based on different methods of treatment.The patients in the study group were given with remove visible intraventricular hematoma by neuroendoscopy,followed by External Ventricular Drainage (EVD) combined with urokinase fibrinolysis.The patients in control group were given with EVD combined with urokinase fibrinolysis.The time of postoperative drainage,ICU stay,duration of onset of fever,the number of intracranial infections,and the proportion of Glasgow outcome scale (GOS) (1 to 5) at 6 months postsurgery were compared between two groups.Measurement data were expressed as (Mean ± SD),and t test was used for measurement data.The count data were analyzed by x2 test or nonparametric rank sum test.Results The time of postoperative drainage,the number of intracranial infections,ICU stay in study group were (6.19 ± 1.1) d,5 cases,(2.8 ± 1.6) d,the indexes in control group were (7.06 ± 1.3) d,15 cases,(5.2 ± 2.0) d.The time of postoperative drainage,ICU stay,the number of intracranial infections were superior to those of the control group,and the difference was statistically significant.The proportion of GOS (1 to 5) at 6 months after surgery was 5 cases (12.2％),5 cases (12.2％),10 cases (24.4％),14 cases (34.1％),7 cases (17.1％) in study group,the indexes in control group were 10 cases(20.0％),13 cases (26.0％),11 cases(22.0％),10 cases(20.0％),6 cases(12.0％).The 6-month postoperative GOS of the study group were superior to those of the control group,and the difference was statistically significant (P ＜ 0.05).Conclusion Neruendoscopy combined with urokinase in the treatment of spontaneous intraventricular hemorrhage can reduce the time of postoperative drainage and the incidence of intracranial infection,shorten the time of ICU stay and improve the functional prognosis of the patients.

Objective:To explore the expression level and clinicopathological characteristics of 14-3-3θ protein in distal cholangiocarcinoma tissues, and further analyze the long-term prognosis of patients with different expression levels.Methods:A retrospective cohort study was conducted to collect and analyze the clinical data of 135 patients with distal cholangiocarcinoma who underwent surgical resection at the Department of Hepatobiliary Surgery, Beijing Chaoyang Hospital Affiliated to Capital Medical University from January 2015 to December 2021, including 86 males and 49 females.(65.1±10.1) years old. Immunohistochemistry was used to detect the expression level of 14-3-3θ protein in cholangiocarcinoma tissue. The postoperative pathological sections of patients were evaluated based on the immune response score(IRS). The optimal cutoff value was determined through the receiver operating characteristic(ROC) curve was 3.5. Currently, the area under the curve was 0.741, the sensitivity was 73.5%, and the specificity was 71.3%. The patients were divided into two groups: IRS<4 was the 14-3-3θ low expression group( n=81), IRS≥4 was the 14-3-3θ high expression group( n=54). After surgery, the patient′s survival status was followed up through a combination of outpatient review and telephone follow-up. Normally distributed measurement data were expressed as mean ± standard deviation( ± s), and comparisons between groups were made using the t test; non-normally distributed measurement data were expressed as M( Q1, Q3), and comparisons between groups were made using the rank sum test. The chi-square test was used to compare enumeration data between groups. Results:The preoperative CA19-9 and lymph node metastasis in the 14-3-3θ low expression group were 44.3(20.8, 132.2) U/mL and 28 cases respectively. The preoperative CA19-9 and lymph node metastasis in the 14-3-3θ high expression group were 82.3(43.4, 396.9) U/mL and 32 cases respectively. The difference between the two groups was statistically significant( P<0.05). All patients had regular postoperative telephone calls or return to the hospital for follow-up. The median postoperative survival time of the 14-3-3θ low-expression group was 36 months. The 1-, 2-, and 3-year survival rates after surgery were 88.9%, 66.5%, and 66.5%, respectively. 49.4%, the median survival time after surgery in the 14-3-3θ high expression group was 13 months, and the 1, 2, and 3-year survival rates after surgery were 53.7%, 23.3%, and 13.3% respectively. The difference between the two groups was statistically significant. significance( P<0.01). Cox proportional hazard model performed multivariate analysis, CA19-9>37 U/mL ( RR=1.970, 95% CI: 1.186-3.272, P=0.009), lymph node metastasis( RR=1.681, 95% CI: 1.035-2.729, P=0.036) and 14-3-3θ staining intensity≥4 ( RR=2.438, 95% CI: 1.546-3.845, P<0.001) have worse long-term prognosis. Conclusions:The expression level of 14-3-3θ protein is related to CA19-9 and lymph node metastasis in distal cholangiocarcinoma. A high expression of 14-3-3θ protein indicates poor long-term prognosis in patients with extrahepatic cholangiocarcinoma.

Objective:To investigate the clinical efficacy and prognostic influencing factors of radical surgery for duodenal gastrointestinal stromal tumor (GIST).Methods:The retrospective cohort study was conducted. The clinicopathological data of 741 duodenal GIST patients who under-went radical surgery in 17 medical centers, including 121 cases in Renji Hospital Affiliated to Shanghai Jiaotong University School of Medicine, 121 cases in Chinese PLA General Hospital, 116 cases in Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 77 cases in Fudan University Shanghai Cancer Center, 77 cases in West China Hospital, Sichuan University, 31 cases in Guangdong Provincial People′s Hospital, 24 cases in Fujian Cancer Hospital, 22 cases in Fujian Medical University Union Hospital, 25 cases in Shengjing Hospital of China Medical University, 19 cases in Xiangya Hospital, Central South University, 23 cases in Zhejiang Cancer Hospital, 17 cases in Liaoning Cancer Hospital＆Institute, 17 cases in the First Affiliated Hospital of Xiamen University, 15 cases in Sun Yat-sen University Cancer Center, 14 cases in the First Affiliated Hospital of Nanjing Medical University, 14 cases in Zhongshan Hospital Affiliated to Xiamen University and 8 cases in General Hospital of Chinese People′s Liberation Army Air Force, from January 2010 to April 2020 were collected. There were 346 males and 395 females, aged 55(range, 17?86)years. Observation indicators: (1) neoadjuvant treatment; (2) surgical and postoperative situations; (3) follow-up; (4) stratified analysis. Follow-up was conducted using outpatient examination or telephone interview. Patients were followed up once every 3?6 months during neoadjuvant therapy and once every 6?12 months after radical surgery to detect tumor recurrence and survival of patient up to April 2022. Measurement data with normal distribution were represented as Mean± SD. Measurement data with skewed distribution were represented as M(range), and comparison between groups was conducted using the Mann-Whitney U test. Count data were described as absolute numbers or percentages, and comparison between groups was conducted using chi-square test or Fisher exact probability. The Kaplan-Meier method was used to draw survival curves and calculate survival rates. Log-rank test was used for survival analysis. The COX regression model was used for univariate and multivariate analyses. Propensity score matching was done by the 1∶1 nearest neighbor matching method, and the matching tolerance was 0.02. Results:(1) Neoadjuvant therapy. Of the 741 patients, 34 cases received neoadjuvant therapy for 8(range, 3?44)months. Cases assessed as partial response, stable disease and progressive disease before the radical surgery of the 34 cases were 21, 9, 4, respectively. The tumor diameter of the 34 patients before the neoadjuvant therapy and before the radical surgery were 8.0(range, 3.0?26.0)cm and 5.3(range, 3.0?18.0)cm, with the regression rate as 31.9%(range, ?166.7% to 58.3%). (2) Surgical and postoperative situations. Of the 741 patients, 34 cases underwent radical surgery after receiving neoadjuvant therapy, and 707 cases underwent radical surgery directly. All the 741 patients underwent radical surgery successfully, in which 633, 102 and 6 cases received open surgery, laparoscopic surgery and endoscopic treatment, respectively. Of the 633 cases receiving open surgery and the 102 cases receiving laparoscopic surgery, cases with surgical resection range as pancreatoduodenectomy (PD) was 238, and cases with surgical resection range as duodenal limited resection, including duodenal wedge resection, distal gastrectomy, segmental duodenal resection, local resection of duodenal tumor or segmental duodenum combined with subtotal gastrectomy, was 497, 226, 55, 204, 12. Of the 741 patients, 131 cases had post-operative complications including 113 cases with grade Ⅰ?Ⅱ complications and 18 cases with ≥ grade Ⅲ complications of the Clavien-Dindo classification. The duration of postoperative hospital stay of the 741 patients was 13(range, 4?120)days. Of the 707 patients receiving direct radical surgery, 371 cases were evaluated as extremely low risk, low risk, medium risk of the modified National Institutes of Health (NIH) risk classification after surgery, and 336 cases were evaluated as high risk in which 205 cases receive postoperative adjuvant imatinib therapy with the treatment time as 24(range, 6?110)months. (3) Follow-up. All the 741 patients were followed up for 58(range, 7?150)months. During the follow-up, 110 patients had tumor recurrence and metastasis. The 1-, 3-, 5-year overall survival rates and 1-, 3-, 5-year disease-free survival rates of the 741 patients were 100.0%, 98.6%, 94.5% and 98.4%, 90.9%, 84.9%, respectively. The 1-, 3-, 5-year overall survival rates and 1-, 3-, 5-year disease-free survival rates of the 707 patients receiving direct radical surgery were 100.0%, 98.5%, 94.3% and 98.4%, 91.1%, 85.4%, respectively. (4) Stratified analysis. ① Analysis of prognostic factors in patients undergoing radical surgery directly. Results of univariate analysis showed that primary tumor location, tumor diameter, mitotic count, modified NIH risk classification and tumor gene information were related factors affecting the overall survival of 707 patients with primary duodenal GIST who underwent direct radical surgery ( hazard ratio=0.43, 0.18, 0.22, 0.06, 0.29, 95% confidence intervals as 0.20?0.93, 0.09?0.35, 0.10?0.50, 0.03?0.12, 0.09?0.95, P<0.05). The primary tumor location, tumor diameter, mitotic count, modified NIH risk classification were related factors affecting the disease-free survival of 707 patients with primary duodenal GIST who underwent direct radical surgery ( hazard ratio=0.65, 0.25, 0.25, 0.10, 95% confidence intervals as 0.41?1.03, 0.17?0.37, 0.15?0.42, 0.07?0.15, P<0.05). Results of multivariate analysis showed that primary tumor located at the horizontal segment of duodenum, mitotic count >5/50 high power field, tumor gene KIT exon 9 mutation were independent risk factors affecting the overall survival of 365 patients with primary duodenal GIST after removing 342 patients without tumor gene information who underwent direct radical surgery ( hazard ratio=2.85, 2.73, 3.13, 95% confidence intervals as 1.12?7.20, 1.07?6.94, 1.23?7.93, P<0.05). Tumor diameter >5 cm and mitotic count >5/50 high power field were independent risk factors affecting the disease-free survival of 707 patients with primary duodenal GIST who underwent direct radical surgery ( hazard ratio=3.19, 2.98, 95% confidence intervals as 2.05?4.97, 1.99?4.45, P<0.05). ② Effect of postoperative adjuvant therapy on prognosis of high-risk patients of modified NIH risk classification. Of the 336 patients evaluated as high risk of the modified NIH risk classification, the 5-year overall survival rate and 5-year disease-free survival rate were 94.6% and 77.3% in the 205 cases with postoperative adjuvant therapy, versus 83.2% and 64.4% in the 131 cases without postoperative adjuvant therapy, showing significant differences between them ( χ2=8.39, 4.44, P<0.05). Of the 205 patients evaluated as high risk of the modified NIH risk classification who received postoperative adjuvant therapy, there were 106 cases receiving postoperative adjuvant therapy <36 months, with the 5-year overall survival rate and 5-year disease-free survival rate were 87.1% and 58.7%, and there were 99 cases receiving post-operative adjuvant therapy ≥36 months, with the 5-year overall survival rate and 5-year disease-free survival rate were 100.0% and 91.5%. There were significant differences in the 5-year overall survival rate and 5-year disease-free survival rate between the 106 patients and the 99 patients ( χ2=13.92, 29.61, P<0.05). ③ Comparison of clinical efficacy of patients with different surgical methods. Before propensity score matching, cases with primary tumor located at bulb, descending, horizontal, ascending segment of duodenum, cases with tumor diameter ≤5 cm and >5 cm were 95, 307, 147, 34, 331, 252, in the 583 patients receiving open surgery with complete clinical data, versus 15, 46, 17, 5, 67, 16 in the 83 patients receiving laparoscopic surgery with complete clinical data, showing no significant difference in the primary tumor location ( χ2=0.94, P>0.05), and a significant difference in the tumor diameter ( χ2=17.33, P<0.05) between them. After propensity score matching, the above indicator were 16, 39, 20, 8, 67, 16 in the 83 patients receiving open surgery, versus 15, 46, 17, 5, 67, 16 in the 83 patients receiving laparoscopic surgery, showing no significant difference between them ( χ2=1.54, 0.00, P>0.05). Cases with postoperative complications, cases with grade Ⅰ?Ⅱ complica-tions and ≥grade Ⅲ complications of the Clavien-Dindo classification, duration of postoperative hospital stay, the 5-year overall survival rate and 5-year disease-free survival rate were 17, 12, 5, 11(range, 5?120)days, 92.0%, 100.0% in the 83 patients receiving open surgery, versus 9, 7, 2, 11(range, 5?41)days, 91.6%, 97.3% in the 83 patients receiving laparoscopic surgery, showing no signi-ficant difference in postoperative complications, duration of postoperative hospital stay, the 5-year overall survival rate and 5-year disease-free survival rate ( χ2=2.91, Z=3 365.50, χ2=3.02, 1.49, P>0.05) between them. There was no significant difference in complications of the Clavien-Dindo classification between them ( P>0.05). ④ Comparison of clinical efficacy of patients with primary tumor located at the descending segment of duodenum who underwent surgery with different surgical resection scopes. Before propensity score matching, cases with tumor diameter ≤5 cm and >5 cm, cases with tumor located at opposite side of mesangium and mesangium were 71, 85, 28, 128 in the 156 patients with primary tumor located at the descending segment of duodenum who underwent PD with complete clinical data, versus 92, 41, 120, 13 in the 133 patients with primary tumor located at the descending segment of duodenum who underwent duodenal limited resection with complete clinical data, showing significant differences between them ( χ2=16.34, 150.10, P<0.05). After propensity score matching, the above indicator were 28, 13, 16, 25 in the 41 patients with primary tumor located at the descending segment of duodenum who underwent PD with complete clinical data, versus 28, 13, 16, 25 in the 41 patients with primary tumor located at the descending segment of duodenum who underwent duodenal limited resection with complete clinical data, showing no significant difference between them ( χ2=0.00, 0.00, P>0.05). Cases with postopera-tive complications, cases with grade Ⅰ?Ⅱ complications and ≥grade Ⅲ compli-cations of the Clavien-Dindo classification, duration of postoperative hospital stay, the 5-year overall survival rate and 5-year disease-free survival rate were 13, 11, 2, 15(range, 9?62)days, 94.2%, 64.3% in the 41 patients with primary tumor located at the descending segment of duodenum who underwent PD with complete clinical data, versus 9, 8, 0, 15(range, 7?40)days, 100.0%, 78.8% in the 41 patients with primary tumor located at the descending segment of duodenum who underwent duodenal limited resection with complete clinical data, showing no significant difference in post-operative complica-tions, the 5-year overall survival rate and 5-year disease-free survival rate ( χ2=0.99, 0.34, 1.86, P>0.05) between them. There was no significant difference in complications of the Clavien-Dindo classification ( P>0.05) and there was a significant difference in duration of postopera-tive hospital stay ( Z=614.50, P<0.05) between them. Conclusions:The clinical efficacy of radical surgery for duodenal GIST are ideal. Primary tumor located at the horizontal segment of duodenum, mitotic count >5/50 high power field, tumor gene KIT exon 9 mutation are independent risk factors affec-ting the overall survival of patients undergoing direct radical surgery and tumor diameter >5 cm and mitotic count >5/50 high power field are independent risk factors affecting the disease-free survival of patients. There is no significant difference in the short-term efficacy and long-term prognosis between patients undergoing open surgery and laparoscopic surgery. For patients with primary tumor located at the descending segment of duodenum, the duration of postoperative hospital stay is longer in patients undergoing PD compared with patients undergoing duodenal limited resection. For patients evaluated as high risk of the modified NIH risk classification, posto-perative adjuvant therapy and treatment time ≥36 months are conducive to improving the prognosis of patients.

Objective:To analyze the clinical characteristics of patients with bisphosphonates related atypical femoral fractures(AFFS), thereby to facilitate early diagnosis.Methods:The clinical manifestations, biochemical indexes, imaging features and treatment follow-up of AFFS patients who were diagnosed in the Department of Osteoporosis and Bone Disease, the Sixth People′s Hospital Affiliated to Shanghai Jiaotong University from 2011 to 2019 were analyzed retrospectively, and the literature was reviewed.Results:A total of 5 cases of atypical bisphosphonate related femoral fractures were collected, all of them were female, with an average age of 68 years. All the 5 patients were treated with alendronate. Three patients were treated with 70 mg/week throughout the course, and two patients were treated with 10 mg/day at first, and changed to 70 mg/week later. The average course of treatment was 8.7 years, ranging from the shortest 5 years to the longest 17 years. Among the 5 cases, the shortest onset time was 3 years after taking medicine, and the longest was 16 years. The clinical features are as follows: all patients had prodromal pain before fracture which was characterized as dull except for case 4. Case 1 was bilateral thigh pain, the rest were unilateral thigh pain, which began to appear within 2-3 years before fracture. X-ray plain film showed thickening of the lateral bone cortex; radionuclide bone scan(ECT) showed active bone metabolism in the affected area. The abnormal manifestations of ECT were earlier than X-ray and MRI. The recognition of these features is helpful to the early diagnosis of AFFS. All 5 patients stopped bisphosphonates immediately, and continued to take calcium tablets. Active vitamin D was added to 4 cases. One case of incomplete fracture was treated conservatively with Teriparatide for one year, which was helpful to deter it from becoming complete fracture. 4 cases of complete fracture were treated with reduction and fixation, and all healed.Conclusion:Long-term use of bisphosphonates can increase the risk of AFFS. Strengthening the risk assessment during use can reduce the incidence of such fractures. Early diagnosis and reasonable treatment can improve the prognosis.