Objective:To compare the antiplatelet effects of vicagrel and clopidogrel in patients with different cytochrome P450 (CYP) 2C19 genotypes.Methods:This is a post-hoc analysis of a phase Ⅱ clinical trial of vicagrel, which included patients with coronary heart disease who underwent percutaneous coronary intervention from August 2018 to June 2019 in 18 centers. Patients were categorized based on the presence of CYP 2C19 *2 or *3 loss-of-function (LOF) alleles into LOF carrier group ( n=111) and non-LOF carrier group ( n=90). Each group included patients received vicagrel 5 mg, 6 mg, 7.5 mg, or clopidogrel 75 mg for 28 days per study protocol. P2Y 12 reaction units (PRU) were measured using VerifyNow at baseline, 6 to 8 hours after loading dose, 7 to 10 days after randomization, and 28 days after randomization and the percentage inhibition of platelet aggregation (%IPA) was calculated. The primary endpoint was %IPA on day 28. Within the patients from the General Hospital of Northern Theater Command, 8 to 12 patients in each study arms were enrolled in a prespecified pharmacokinetic sub-study, measuring the time to reach maximum plasma concentration (T max), peak plasma concentration (C max), and area under the plasma concentration-time curve (AUC). Results:Among 201 patients, the age was (58.8±8.5) years, and 139 (69.2%) were male. In non-LOF carriers, there was no significant differences in PRU values and %IPA between the vicagrel 5 mg, 6 mg, 7 mg, and clopidogrel groups at all time points (all P>0.05). In LOF carriers, %IPA was significantly higher in the vicagrel-treated groups than in the clopidogrel group at 6-8 hours after loading dose (22.9 (14.2, 31.5)% vs. 19.8 (11.0, 28.6)% vs. 29.5 (20.9, 38.0)% vs. 12.9 (3.9, 21.9)%, P=0.038) and 7-10 days after randomization (22.4 (14.2, 30.5)% vs. 34.4 (26.1, 42.6)% vs. 39.8 (31.8, 47.9)% vs. 24.7 (16.3, 33.2)%, P=0.001), with a trend towards higher %IPA in the vicagrel-treated groups at day 28 (30.4 (21.3, 39.6)% vs. 36.5 (27.2, 45.7)% vs. 40.8 (31.8, 49.8)% vs. 30.7(21.2, 40.2)%, P=0.056). Pharmacokinetic results of 35 patients showed that the C max and AUC of the active metabolite M15-2 of vicagrel was similar to that of clopidogrel in non-LOF carriers, but AUC between vicagrel 5 mg, 6 mg, 7 mg and clopidogrel were significantly different in LOF carriers ((5.6±0.6) h·μg -1·L -1 vs. (6.8±2.7) h·μg -1·L -1 vs. (9.2±3.3) h·μg -1·L -1 vs. (4.2±1.9) h·μg -1·ml -1, P=0.020). Conclusion:Vicagrel and clopidogrel have similar antiplatelet effects in non-LOF carriers, but vicagrel exhibits superior antiplatelet effects in LOF carriers.

Purpose To explore the value of the new generation tau PET tracer 18F-Florzolotau in Alzheimer's disease(AD)at different stages.Materials and Methods Twenty-five MCI patients and sixty-one AD patients with positive β-amyloid status in Huashan Hospital,Fudan University from February 2020 to January 2022 were retrospectively enrolled with 18F-Florzolotau PET imaging and demographic and clinical data.The pre-processed PET images were analyzed by SPM two-sample t-test between MCI and AD groups,and the standardized uptake value ratios(SUVR)were extracted from the region of interest defined by SPM analysis(P＜0.001);scaled subprofile model/principal component analysis was used to construct the different tau related patterns(MCItauRP,ADtauRP)and calculate the corresponding expression values.The classification efficiency of SUVR and MCItauRP,ADtauRP expression values was evaluated by receiver operating characteristic curve.Results Compared with MCI patients,tau protein deposition of AD patients was increased mainly in the bilateral temporal,occipital lobe(P＜0.001),and the SUVR of these brain region in the AD group was higher than that in the MCI group(Z=-3.164,P＜0.00l);the expression values of MCItauRP and ADtauRP were significantly different between the AD group and MCI group(t=3.72,Z=-3.51;both P＜0.001),and these expression values of AD patients were higher than those in the MCI group;the accuracy of tauRP expression values and SUVR for the differentiation between the AD and MCI group were 61.63％,65.12％and 65.12％,respectively;the sensitivity was 88.00％,96.00％and 100.00％,respectively;the specificity was 50.82％,52.46％and 50.82％,respectively.Conclusion The new tau PET can identify and distinguish the differences in tau protein deposition between AD and MCI patients.However,the classification and diagnosis efficiency is not high.In the future,it is necessary to find a more ideal analysis method.

Objective:To explore the efficacy and safety of immunoneoadjuvant therapy with pembrolizumab combined with chemotherapy in locally advanced resectable hypopharyngeal squamous cell carcinoma patients.Methods:This study was a prospective, single arm, single center clinical study that was opened for enrollment in April 2021. Patients who met the inclusion criteria at the Cancer Hospital of the Chinese Academy of Medical Sciences were treated with neoadjuvant therapy of pembrolizumab combined with cisplatin and paclitaxel, and after treatments, received surgery and postoperative adjuvant therapy. The main endpoint of this study was postoperative pathological complete response (pCR), and other observations included adverse reactions and long-term prognoses of patients after neoadjuvant therapy.Results:By September 2023, a total of 23 patients who underwent neoadjuvant therapy and surgery were enrolled in the study and all patients were males aged 49-74 years. All patients were locally advanced stage, including 3 patients in stage Ⅲ and 20 patients in stage Ⅳ. There were 12 cases of primary lesions with posterior ring involvement accompanied by fixation of one vocal cord and 20 cases of regional lymph node metastases classified as N2. Eighteen cases received a two cycle regimen and 5 cases received a three cycle regimen for neoadjuvant therapy. The postoperative pCR rate was 26.1% (6/23), with no surgical delay caused by adverse drug reactions. The laryngeal preservation rate was 87.0% (20/23). Pharyngeal fistula was the main surgical complication, with an incidence of 21.7% (5/23). The median follow-up time was 15 months, and 3 patients experienced local recurrence.Conclusions:The immunoneoadjuvant therapy of pembrolizumab combined with chemotherapy has a high pCR rate in locally advanced resectable hypopharyngeal squamous cell carcinoma, with increased laryngeal preservation rate and no significant impact on surgical safety.

Multi-centre clinical trials on PET/CT brain imaging are complex to organize and require careful co-ordination and management. This article describes considerations, which are necessary when designing and starting a multi-centre clinical trial on PET/CT brain imaging, based on guidelines and multi-center clinical brain imaging studies, providing references for further studies.

Objective To explore the function and survival of islet grafts during co-transplantation with adipose mesenchymal stem cells (AMSCs) in diabetic mice .Methods After human AMSCs and islet cells were isolated ,purified and then subcutaneously co-transplanted into nude mice with diabetes mellitus . Four groups of AMSCs + islet co-transplantation , islet transplantation alone ,phosphate buffered solution (PBS) and normal control mice were designated . Islet cell activity and apoptosis/revascularization degree of islet grafts were observed by immunohistochemical double staining of insulin ,factor associated suicide (Fas) and CD31 antibody . The blood glucose and serum insulin levels of mice and the survival time of islet grafts were compared . Results The blood glucose and serum insulin levels of diabetic mice analyzed by multivariate analysis in AMSCs + islet co-transplantation group were better than those in islet transplantation alone group (P< 0 .05 ) . The mean survival time (MST ) of islet grafts was longer in AMSCs + islet co-transplantation group than that in islet transplantation alone group [(81 .33 ± 7 .58) vs .(58 .17 ± 6 .91) days] (P<0 .05) .At Day 7 post-transplantation ,insulin staining intensity of islet grafts was higher in AMSCs + islet co-transplantation group than that in islet transplantation alone group while Fas staining intensity of islet grafts was lower .And mean microvascular density (MVD) of islet grafts per square millimeter was higher in AMSCs + islet co-transplantation group than that in islet transplantation alone group [(21 .8 ± 5 .6 ) vs . (14 .6 ± 4 .1 )] ( P< 0 .05 ) .Conclusions Co-transplantation with AMSCs may improve the function of islet grafts ,prolong its survival and promote its revascularization .

Objective To investigate the revascular and transplanted effects of islet grafts after co-transplantation with vascular endothelial cells ( ECs) in diabetic rats. Methods The rat ECs and islet cells were isolated and purified, then subcutaneously co-transplanted to the inbred SD male rats with diabetes mellitus, the group of islets transplanted alone, group of phosphate buffered solution, and group of normal rats served as control. Islet grafts revascularization degree, islet cells activity and apoptosis were observed by immunohistochemical double staining of CD31, insulin and factor associated suicide (Fas) antibody. The results of intraperitoneal glucose tolerance test (IPGTT) and intraperitoneal insulin tolerance test (IPITT), the blood glucose and insulin levels of the rats and the survival time of the islet grafts were compared. Results The blood glucose concentrations of IPGTT and IPITT, the blood glucose and insulin levels of rats analyzed by multivariate analysis in the ECs and islets transplantation group were better than those in the islets transplantation alone group (P<0.05). The islet grafts mean survival time of the ECs and islets transplantation group was longer than that of the islets transplantation alone group (P<0.05). On the 7th day after transplantation, mean microvascular density of islet grafts per square millimeter in the ECs and islets transplantation group was 26.4 ± 6.1, significantly greater than that in the islets transplantation alone group 18.3 ± 5.7 (P<0.05). In the ECs and islets transplantation group, insulin staining intensity was higher than that in the islets transplantation alone group (P<0.05), while factor associated suicide(Fas) staining intensity was lower than that in the islets transplantation alone group ( P<0. 05 ). Conclusion Co-transplantation with ECs could promote the revascularization of islet grafts and improve the effect of islet transplantation.

Objective To investigate the influence of VEGF165 gene transfection on the revascularization and transplantation effect of islet grafts.Methods The rat islet cells were transfected with lentivirus containing VEGF165 gene (LV-VEGF165) in vitro,then transplanted to the renal capsule of inbred SD male rats with diabetes mellitus.Islet cells transfected with lentivirus marked with AcGFP1 and no VEGF165 gene were set as null vector control group (LV-AcGFP1 group),individual islet cells as blank control group.VEGF165 expression in vivo and in vitro was detected by ELISA.The islet grafts revascularization degree and islet cell activity were observed by immunohistochemical double staining of insulin and CD31 antibody.Blood glucose and insulin level of rats with diabetes mellitus and survival time of islet grafts were compared.Results The VEGF165 concentrations in the LV-VEGF165 group secreted by islet cells in vivo and in vitro were significantly higher than those in the LV-AcGFP1 group and the blank control group (P＜0.01).Mean microvascular density (MVD) of islet grafts per square millimeter in the LV-VEGF165 group was (24.3 ± 3.7),significantly greater than that in the LV-AcGFP1 group (12.4 ± 2.5) and the blank control group (12.4 ± 2.5) (P＜ 0.01).Insulin staining intensity in the LV-VEGF165 group was also higher than that in the LVAcGFP1 group and the blank control group.The blood glucose and insulin levels in rats with diabetes mellitus analyzed by multivariate analysis in the LV-VEGF165 group were controlled more effectively than those in the LV-AcGFP1 group and the blank control group (P＜0.01).The islet grafts mean survival time (MST) after transplantation in the LV-VEGF165 group was longer than that of the LVAcGFP1 group and the blank control group (P＜0).01).Conclusion The VEGF165 gene transfection to islet cells could promote the revascularization of islet grafts and improve the effect of islet transplantation.

ABSTRACT:Objective For preparation of vascularized islets , to isolate and culture human adipose derived stem cells , investigate the role of adipose derived stem cells (ADSCs ) in promoting the proliferation and vascularization of human umbilical vein endothelial cells (HUVECs ) co‐cultured in vitro , and explore its mechanism .Methods ADSCs and HUVECs were isolated by collagenase digestion method ,then cultured ,and identified by morphology ,immunofluorescence or multi‐directional differentiation .The co‐culture system of ADSCs and HUVECs was established , HUVECs cultured alone were set up for control group . The proliferation , vascularization and concentration of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (b‐FGF)in the supernatant were compared between the two groups .Results The third generational ADSCs had uniform long spindle fiberous morphology and multi‐directional differentiational function . Immunofluorescence test of surface antigens on ADSCs revealed CD44/CD49d (+ ) ,CD31/CD34 (-) ,on HUVECs CD31/vWF (+ ) . High vascular density was found when co‐cultured in Matrigel of ADSCs and HUVECs than alone of HUVECs .Growth curve shown at days 3 , 4 and 5 of the logarithmic phase , HUVECs count in co‐culture group of ADSCs and HUVECs was (4 .52 ± 0 .31) × 104 ,(7 .18 ± 0 .45) × 104 ,and (8 .23 ± 0 .36) × 104 under indirect co‐culture condition , while that in individual HUVECs group was (2 .71 ± 0 .25) × 104 ,(4 .87 ± 0 .26) × 104 ,and (6 .86 ± 0 .33) × 104 ( P<0 .01) .Population doubling time of HUVECs was shorter in co‐culture group than in individual group .Also ,the OD value of HUVECs was higher in co‐culture group than in individual group when cultured at days 1 ,3 ,5 and 7 ( P<0 .01) .When cultured at days 3 ,7 and 13 ,the concentration of VEGF and b‐FGF in the supernatant was higher in co‐culture group than in individual group ( P< 0 .01 ) . Conclusion ADSCs can promote the proliferation and vascularization of HUVECs in vitro co‐culture conditions by secreting or increasing the HUVECs secretion of VEGF and b‐FGF .

Objective To observe the effects of the intervention of Didang Decoction at different times on changes of AMPK signaling pathway related factors in macrovascular endotheliocytes of diabetic rats; To discuss the mechanism of mitochondria energy metabolism regulating the AMPK signaling pathway for macrovascular endothelial defense function. Methods Injection of STZ into the caudal vein and administration of high fat diet wer used to generate diabetic rat model. All rats were randomly divided into the following 7 groups: control, model, metformin, simvastatin, early-, middle-, and late-stage Didang Decoction group. Western blot was used to detect the expressions of APMKα1 and PGC-1α in rat aortic endothelial cells. Changes in the intracellular AMP and ATP levels were detected by ELISA. Real time fluorescence quantitative PCR was used to detected mRNA expressions of Caspase-3, eNOS, and Bcl-2 in tissue of thoracic aorta. Results Compared with the model group, the expressions of AMPKα1 and PGC-1α in the early-stage and middle-stage Didang Decoction group and simvastatin group increased (P<0.05); the gene expressions of Bcl-2, and eNOS significantly increased in the early-stage Didang Decoction group and simvastatin group (P<0.05), while the expressions of Caspase-3 decreased significantly (P<0.05). The expression of ATP increased significantly and the expression of AMP decreased significantly in the early-stage Didang Decoction group and simvastatin group (P<0.05), and the best effects were shown in the early-stage Didang Decoction group. Conclusion Early intervention of Didang Decoction can enhance energy metabolism in the mitochondria of macrovascular endothelial cells by regulating the AMPK signaling pathway, and then plays a role in strengthening the defense function of macrovascular endothelial cells.

Objective To isolate and culture human adipose-derived stem cells (hADSCs),investigate the influence of hADSCs on the cellular morphology,survival rate,and function of human islet cells under the in vitro non-contact co-culture conditions,and explore its mechanism.Method hADSCs were isolated by collagenase digestion method,then cultured,and identified by morphology,immunofluorescence and multi-directional differentiation.Adult islet cells were separated and purified by Liberase enzyme and Ficoll 400,then divided into co-culture group and individual group.The cellular growth morphology of islet cells was observed by inverted phase contrast microscope.The survival rate of islet cells,insulin secretory volume,insulin stimulation index and concentration of growth factor in the supernatant were compared between the two groups.Result hADSCs of the third generation showed uniform long spindle fibrocyte-like morphology,and had multi-directional differentiational potentials of osteogenesis and adipogenesis.Immunofluorescence test of surface antigens on hADSCs revealed CD44 + and CD49d +,CD31-,CD34-and CD106-.After 14-day culture,the islet cellular morphology in co-culture group was more intact than that in individual group.The survival rate of islet cells in co-culture group was (82.83 + 2.32) ％,and that in individual group was (53.00 + 2.82) ％ (P＜0.01).Insulin secretory volumes were (23.66 + 2.11) and (7.82 +1.09) mU/L respectively in co-culture group and individual group under high glucose concentration,and 13.22 + 0.77 and 6.40 + 0.44 mU/L respectively under low glucose concentration (P＜0.01 for all).Insulin stimulation index was decreased from 1.67 + 0.10 (at 3rd day) to 1.77 + 0.13 (at 14th day) in co-culture group,and from (1.67 + 0.10) (at 3rd day) to (1.77 + 0.13) (at 14th day) in individual group (P＜0.01).After 14-day culture,the concentrations of HGF,TGF-β,VEGF and bFGF in the supernatant were higher in co-culture group than in individual group (P＜0.01).Conclusion hADSCs were isolated and cultured successfully from adult adipose tissue.They could increase the survival rate and improve the function of islet cells when co-culture with the adult islet cells in vitro through secreting HGF,TGF-β,VEGF and b-FGF.