Objective To develop and verify a cation exchange high performance liquid chromatography(CEX-HPLC)method for the detection of charge variants of pembrolizumab.Methods Pembrolizumab was bound to the exchange column matrix by using MabPac SCX-10 column,and the variants with different charges were eluted by gradually increasing the salt concentration of the mobile phase.The specificity,precision,linear range,accuracy and durability of the method were verified,and the charge variants of three batches of pembrolizumab finished products were detected by using the developed method.Results The resolution of the last acidic isomer peak and the first basic isomer peak of pembrolizumab from the main peak were 1.28 and 1.42,respectively.The mobile phase A and preparation buffer had no obvious interference peaks at the peak of the sample;The RSD values of the precision verification were all less than 2.0%;The total peak area,main peak area,acidic isomer peak area and basic isomer peak area of the standard all exhibited good linear relationship with the theoretical dilution concentration with each R~2 of 1.00;The recovery rates of the total peak area and main peak area of the standard at three concentrations were between 96.81% and 106.07%;When pH value of the mobile phase was within the range of 6.30±0.10,the RSD values of the total peak area and main peak area percentage of the standard were1.5% and 1.9%,and when the column temperature was within the range of(35±4) ℃,the RSD values of the total peak area and main peak area percentage of the standard were 0.4% and 0.3%,respectively.The RSD value of the main peak retention time of the three batches of finished products was 0.Conclusion The developed CEX-HPLC method can effectively separate the acidic isomers,main peaks and basic isomers of pembrolizumab with good specificity,precision and accuracy,which can be used for the follow-up research and development of pembrolizumab,the process verification of expanding production and the stability research.

BACKGROUND: It was uncertain that the migration, differentiation and survival of transplanted bone marrow mesenchymal stem cells (BMSCs) into myocardium after the acute myocardial infarction. OBJECTIVE: To investigate the migration, differentiation and survival of rabbit transplanted autologous BMSCs in myocardium after the acute myocardial infarction. METHODS: Rabbit BMSCs were isolated and labeled by DAPI in vitro. Rabbit left anterior descending branch was ligated to establish acute myocardial infarction models. Following successful model establishment, 30 New Zealand rabbits were assigned to BMSC and control groups (n = 15). In the BMSC group, autologous BMSCs were infused into the surrounding sites of the infracted region by 4 points 1 hour following coronary artery ligation. In the control group, the same region was injected with an equal volume of saline. Injection volume was 30 μL in each point. Five animals from each group were sacrificed 10 minutes, 3 days and 4 weeks following transplantation. The heart was obtained to undergo frozen sections. The distribution of DAPI-labeled BMSCs was observed using fluorescence microscope. Immunofluorescence method was used to examine the troponin Ⅰ and α-actin. RESULTS AND CONCLUSION: DAPI-labeled BMSCs with blue nuclei were distributed extensively in the myocardium of the cell transplantation group, ovoid in shape and arranged in parallel with the cardiac muscle fibers. Troponin Ⅰ and α-actin were positive immunofluorescently in the cytoplasm of the labeled BMSCs. Results indicated that transplanted BMSCs in the ischemic myocardium could differentiate into myocardial cells under stimulation of local microenvironment.

Objective To further prove the multipotentiality of rabbit bone mesenchymal stem cells (MSCs) by inducing it into cardiomyocytes with 5-azacytidine. Methods MSCs from rabbit were abstracted, isolated, cultured. Then MSCs were induced by 5-azacytidine in vitro for 24 hours. After cultured for 4 weeks, MSCs’ differentiation was studied by immunohistochemistry and electron microscopy technique. Results Some of MSCs induced by 5-azacytidine expressed troponinT . Electron microscopy showed myofilaments. Conclusion MSCs which exist in bone marrow are mutipotential stem cells, and they can be induced into cardiomyocytes by 5-azacytidine in vitro.

Objective. To evaluate the respective value of dual-phase helical CT arterial portography (CTAP) and conventional angiography in preoperative predicting resectability of pancreatic ductal adenocarcinoma. Subjects and methods. Tumor resectability was prospectively evaluated in 54 patients with pathologically proven pancreatic ductal adenocarcinoma who later underwent surgery. Both dual-phase helical CT scanning and selective angiography were obtained in each patient preoperatively. For optimal enhancement of pancreas and major peripancreatic vessels, two catheters connected to an automatic injector via a Y-shaped tube were placed after selective angiography,one in celiac trunk, the other in superior mesenteric artery. Then the patient underwent dual-phase helical CTAP of pancreas and liver. The criteria of irresectability for CTAP include: tumor diameter≥ 5 cm,extrapancreatic invasion, distant metastases and vascular involvement(occlusion, stenosis or semicircular encasement of superior mesenteric artery, hepatic artery, splenic artery, celiac axis; portal vein, superior mesenteric vein or splenic vein). The results of both modalities were correlated with findings from surgery or pathology. Results. Thirty-eight of 54 patients had nonresectable disease. In prediction the irresectability, sensitivity, specificity, positive predictive value, negative predictive value and overall accuracy were 94.7％ ,100％ ,100％ ,88.9％ ,96.3％ respectively for helical CTAP and 63.2％ ,93.8％ ,96.0％ ,51.7％ ,72.2％ respectively for selective angiography. In assessing vascular involvements, dual-phase helical CTAP was also superior to selective angiography. Conclusion. Dual-phase helical CTAP is superior to angiography in assessing resectability of pancreatic ductal adenocarcinoma. The combination of the two modalities may further improve overall accuracy of assessment.

Objective To determine the effecting factors for diagnosis of binign or malignant in cystic lesions of pancreas(CLP).Methods One hundred twenty-six patients undergoing operations for CLP or suspected CLP in this hospital from January 1984 to June 2008 were reviewed.Patients were divided into two groups according to lesion's histological features after operation.The predictive effect of various preoperative factors on the malignant potential of CLP was evaluated.Results One hundred twenty-six patients underwent operations for suspected pancreatic cystic neoplasms.There were 89 benign and 37 malignant CLPs.The univariate analysis showed that gender, clinical symptoms(jaundice and weight loss), elevated serum CA199, and presence of one or more of three morphologic features such as solid component, nodule or septation were significantly different between benign and malignant cystic neoplasm of pancreas.The multivariate analysis indicated that imaging features and gender were independent predictors of malignancy.Conclusion In patients with suspected pancreatic cystic neoplasms, elevated serum CA199, clinical symptoms(jaundice and weight loss)and presence of suspicious morphologic features on imaging are predictors of malignant potential of CLP.Patients with a high likelihood of a potentially malignant or malignant lesion based on these three factors should undergo operation without additional investigations.

Objective To summarize the diagnostic and therapeutic experiences for multiple insulinoma. Methods Clinical data of 34 cKsefl of multiple insulinoma treated in Peking Union Medical College Hospital between 1984 and 2007 were analyzed retrospectively. Results Multiple insulinoma was identified in these 34 cases for 37 instances.Malignant insulinoma was found in 2 cases.Three cases suffered from postoperative recurrent multipie tumors.35.3% cases belonged to MEN1;13.5% cases were of insulinoma combined with islet hyperplasia;43.2% cases had 3 or more than 3 insulinomas;Fifteen cases (40.5%)had had a misdiagnosis.45.2%tumors were smaller than 1 cm in diameter:88.9%multiple insulinonla located at the body and tail of the pancreas.Enucleation of multiple tumors was performed for 48.7 percent of cases. Conclusions Most multiple insulinomas were small,it was difficult for preoperative examination to locate all the tumors therefore.Being on the alert against multiple insulinoma and such measures as careful exploration,intraoperative blood glucose determination.fine needle aspiration biopsy,frozen sections helps to avoid missing multiple imuhnoma.

Objective To investigate the Effects of inflammatory response and postoperative analgesia on thoracic paravertebral nerve block in patients undergoing radical resection of lung cancer. Methods In this study, 68 patients with radical resection of lung cancer were divided into observation group(34 cases)and control group (34 cases)randomly. The anesthesia method of the observation group was general anesthesia combined with thorac-ic paravertebral nerve block ,and the anesthesia method of control group was general anesthesia. Serum IL-6 and IL-10 concentrations were measured before anesthesia(T0),30 min after surgery starts(T1),surgery end(T2),6h after surgery(T3). The visual analogue score of 2 h,12 h,24 h and 48 h postoperatively in two groups were re-corded. The postoperative 24h sufentanil dosage,the number of PCIA pump pressure and the total amount of back-ground infusion were measured. Results Compared with the control group,the serum IL-6 level of T2 and T3 in the observation group were significantly lower than those of the control group ,while the IL-10 concentration was significantly higher in the observation group(P<0.05). The VAS scores of 2 h,12 h and 24 h in the observation group were significantly lower than those in the control group(P<0.05). The 24h sufentanil dosage,PCIA pump pressure and the total amount of background infusion in the observation group were significantly lower than those in the control group(P < 0.05). Conclusions Preoperative thoracic paravertebral nerve block can significantly re-duce the inflammatory response and enhance postoperative analgesia.

Objective To summarize our experience with surgical management of renal neoplasm involving inferoir veno cava.Methods We review the data of 115 patients,including 74 male patients and 41 female patients,with renal neoplasm involving venous system between March 1993 and December 2015.The mean age was 52 years old,ranging 22 to 77 years old.The tumor was found in right side in 77 cases and left side in 38 cases.There were 15 patients (13.0％) with renal vein thrombus,38 (33.1％) with infrahepatic thrombus,29 (25.2％) with low retrohepatic thrombus,20 (17.4％) with high retrohepatic thrombus,and 13 (11.3％) with supradiaphragmatic thrombus.The mean age was 52 years old,ranging 22 to 77 years old.The tumor was found in right side in 77 cases and left side in 38 cases.All patients accepted the radical nephrectomy.Primary outcomes were overall survival (OS),and cancer special survival (CSS) in patients with renal cell carcinoma (RCC) estimated by Kaplan-Meier method.Secondary outcome included operative and oncological features,past-operative complications and hospital mortality.Cox proportional hazard model was used to univariate and multivariate analysis for risk factor impacting on OS of RCC patients.Results Complete resections of renal neoplasm with tumor thrombus were achieved in 113 patients (98.3％),2 patients died intraoperatively due to the dropping of thrombus.Postoperative complication rate was 16.8％.Hospital mortality was 2.6％.Mean follow-up interval was 48 months.OS rates at 5-,and 10-year in RCC patients were 66.5％,and 61.8％,respectively.Metastasis was an independent risk factor affecting on OS (P =0.000).However,the level of thrombus was not an risk factor affecting the prognosis.Conclusions Radical resection of renal tumor and caval thrombus is an effective treatment for prolonging survival in patients with RCC extending into venous system.Retrohepatic caval tumor thrombus below or above the main hepatic vein should be dichotomized and managed respectively with diverse techniques.Metastatic rather than the level of thrombus was a risk factor impacting on RCC patient survival.

Objective: To explore the effect of enterostomy on analgesic pattern in advanced digestive tract cancer. Methods: A retrospective cohort study was carried out, which was approved by the Ethics Committee of the Sixth Affiliated Hospital of Sun Yat-sen University (E2018026). Inclusion criteria were as follows: (1)age and gender were not limited; (2) all the gastrointestinal malignancies were confirmed histologically, and local recurrence or metastasis were confirmed by CT or MR; (3) numerical rating scale (NRS) ≥4 points, opioid analgesic drugs were required; (4) informed consents were signed by patients of their own. Exclusion criteria were as follows: (1) malignancies of early stage; (2) suspicious adverse mental states which might lead to poor administration compliance; (3) hypersensitivity or allergic reactions to opioids. Clinical data of patients with advanced gastrointestinal cancer receiving comprehensive treatment at the Medical Oncology Department of the Sixth Affiliated Hospital of Sun Yat-sen University from September 2016 to April 2017 were retrospectively collected. The patients were divided into the stoma group and the non-stoma group. The clinical findings of two groups were analyzed, including age, sex, ostomy status, pain location, presence or absence of intestinal obstruction, pain characteristics, selection of opioid analgesic agents, treatment of side effects of analgesics. Pain was assessed using brief pain inventory(BPI) table and NRS score. Strong opioids were prescribed for patients of NRS ≥4. Patients who were intolerant to opioids required opioid titration. The titration drugs included oral or IV morphine and oxycodone. After achievement of adequate pain control, long-acting opioids were administered, which included sustained-release morphine tablets, controlled release oxycodone and transdermal fentanyl. Criteria for pain relief included NRS≤3, breakthrough pain <3 times/day and duration of adequate pain control >3 days. The χ² test and the Wilcoxon signed rank sum test (nonparametric test) were used to analyze the clinical features of patients in the stoma and non-stomach groups. In order to find the factors associated with maintenance therapy and the use of laxatives, the variables were compared as well as in multivariate analysis with multiple regression models. For all the statistical tests, a value of P<0.05 in a two-tailed test was established as the alpha significance level. Result: A total of 123 patients were enrolled in this study, including 79 males (64.2%) and 44 females (35.8%) with a median age of 51 years. Fifty-two patients were in stoma group, including 30 (24.4%) of ileostomy and 22 (17.9%) of colostomy, and 71 patients were in non-stoma group. Pain of 40 (76.9%) patients in stoma group located in abdomenopelvic site while the pain of 44 (62.0%) patients in non-stoma group located in other sites. Compared with non-stoma group, cases in stoma group complained more abdominopelvic pain (73% vs. 62.0%, P<0.001).The median NRS pain score before treatment in the stoma group and the non-stoma group was 5.7 and 5.6, respectively, without statistically significant difference (P=0.741). After analgesic management, the above scores reduced to 2.1 and 2.3, respectively, without statistically significant difference as well (P=0.092). Analgesic treatment was effective in 111 cases (90.2%), including 49 cases (94.2%) in the stoma group, and 62 cases (87.3%) in the non-stoma group, and there was no statistically significant difference between the two groups (P=0.202). There was more application of fentanyl transdermal patch [34.6%(18/52) vs. 9.8%(7/71)] in the stoma group, while more application of lactulose laxative [78.9%(56/71) vs. 61.5%(32/52)](χ²=10.023, P=0.002) in the non-stoma group. Multivariate analysis revealed that ostomy (OR=0.290, 95%CI: 0.102-0.824, P=0.009) and pain site (OR=5.691, 95%CI:1.709-18.948, P=0.005) were independent factors affecting the choice of the first line opioid sustained release agent. Of the 123 patients with maintaining analgesia, 98 had available data of laxative use, of whom 46 used laxatives to prevent or treat constipation, and the proportion of laxatives in stoma group (21.2%, 11/52) was significantly lower than that in non-stoma group (49.3%, 35/71) (χ²=6.957, P=0.008). Multivariate analysis of the application of laxative use showed that age (OR=0.281, 95% CI: 0.123-0.684, P=0.010) and ostomy (OR=2.621, 95% CI: 1.033-6.687, P=0.045) were independent factors affecting the use of lactulose laxatives. Conclusions Enterostomy may affect the analgesic pattern in advanced digestive tract cancer. Patients with stoma are more likely to use fentanyl transdermal patches and younger patients with stoma do not need prophylactic use of laxatives.

The STandards for Reporting Interventions in Clinical Trials Of Moxibustion (STRICTOM), in the form of a checklist and descriptions of checklist items, were designed to improve reporting of moxibustion trials, and thereby facilitating their interpretation and replication. The STRICTOM checklist included 7 items and 16 sub-items. These set out reporting guidelines for the moxibustion rationale, details of moxibustion, treatment regimen, other components of treatment, treatment provider background, control and comparator interventions, and precaution measures. In addition, there were descriptions of each item and examples of good reporting. It is intended that the STRICTOM can be used in conjunction with the main CONSORT Statement, extensions for nonpharmacologic treatment and pragmatic trials, and thereby raise the quality of reporting of clinical trials of moxibustion. Further comments will be solicited from the experts of the CONSORT Group, the STRICTA Group, acupuncture and moxibustion societies, and clinical trial authors for optimizing the STRICTOM.
Clinical Trials as Topic ; methods ; standards ; Humans ; Moxibustion ; methods ; standards ; Randomized Controlled Trials as Topic ; Research Design ; standards