BACKGROUND:Studies have shown that the number of osteoblasts is often decreased after osteoporosis, and osteoblast replacement therapy becomes a new target for the treatment of osteoporosis. OBJECTIVE:To observe the osteogenic differentiation of human bone marrow mesenchymal stem cels cultured in dexamethasone, vitamin C and beta-glycerophosphate. METHODS:Mesenchymal stem cels were isolated and purified from adult bone marrow using human lymphocyte separation medium. The expression of cel surface markers was detected by flow cytometry. Cel ultrastructure was observed by transmission electron microscope. Then, the bone marrow mesenchymal stem cels were cultured in osteogenic induction medium containing dexamethasone, vitamin C andβ-glycerophosphate, and RT-PCR was used to detect the bone morphogenetic protein-2 mRNA expression after osteogenic induction. RESULTS AND CONCLUSION:A large number of adherent cels were visible as fibrous growth at 2 weeks after culture and strongly expressed CD44, CD29, but did not express CD34, CD45. These cels could be induced to differentiate into osteoblasts, and express bone morphogenetic protein-2 mRNA. Alizarin red staining and alkaline phosphatase staining were positive for the cels. These findings suggest that human bone marrow mesenchymal stem cels cultured in dexamethasone, vitamin C and beta-glycerophosphate can differentiate into osteoblasts, and has a potential for the treatment of osteoporosis.

BACKGROUND:The main clinical manifestation of senile arteriosclerosis obliterans is lower limb ischemia, which is currently difficult to treat. One method is by autologous stem cell transplantation into the muscles of ischemic limbs to improve the formation of new capillaries and restore lower limb blood flow. Endothelial progenitor cell marker CD34+ cell transplantation has been shown to promote angiogenesis in ischemic limbs. Therefore, we propose that peripheral blood autologous CD34+ cell transplantation in older adult patients with atherosclerotic ischemia could effectively promote angiogenesis.OBJECTIVE:To assume that peripheral blood autologous CD34+ cell transplantation in the elderly with atherosclerotic ischemia could effectively promote angiogenesis.METHODS:This is a prospective, single-center, open-label, randomized, and controlled clinical trial that will be completed at the Qingdao No. 9 People's Hospital, China. Twenty older adult patients with atherosclerotic lower limb ischemia will be randomized into two groups. In the cell transplantation group (n=10), peripheral blood CD34+ cells transfected with vascular endothelial growth factor 165 (VEGF165) gene will be intramuscularly transplanted into the ischemic limbs in older adult patients with atherosclerotic lower limb ischemia. In the control group (n=10), normal saline will be intramuscularly injected into the ischemic limbs. All patients will be followed up for 6 months. The primary outcome will be ankle-brachial indices before and 6 months after transplantation to assess lower limb ischemia in both groups.The secondary outcomes will be the number of microvessels in the lower limb muscles before and 6 months after transplantation, the morphology of new blood vessels revealed by CT angiography, the number of VEGF-immunoreactive cells 6 months after transplantation and the incidence of adverse reactions. The trial was registered at the ClinicalTrials.gov (identifier:NCT03098771), and the study protocol was approved by the Ethics Committee of Qingdao No. 9 People's Hospital of China. All protocols will be in accordance with Declaration of Helsinki,formulated by the World Medical Association. All patients will be informed of study protocols and provide a written informed consent prior to the beginning of the trial.DISCUSSION:This trial will begin in January 2018 and finish in December 2019. We aim to quantify the effects of VEGF165 gene-modified CD34+ cell transplantation in the treatment of older adult patients with atherosclerotic ischemia to develop a new effective treatment of lower limb ischemia.

Objective To explore the efficacy and safety of bifidobacterium tetragenous viable bacteria tablets (BTVBT) in blood glucose control in patients with type 2 diabetes mellitus (T2DM).Methods This study was a randomized, double-blind, placebo parallel comparison, multicentre clinical research.The subjects were T2DM patients who were using anti-hyperglycemic drugs.They were randomly divided into observation group and control group according to 1∶1 ratio.The subjects accepted the therapy of BTVBT or placebo by oral administration (3 tablets, tid) for eight weeks, followed up for 4 weeks, during which the basic treatment maintained unchanged.The primary outcomes: the changes of glycosylate hemoglobin A1c (HbA1c) from baseline.Results Totally 234 subjects (116 cases in observation group and 118 cases in control group) from 7 centers were included in the study.The baseline characteristics were comparable between these two groups.The HbA1c was (8.00±1.08)％ and (7.99±1.03)％ in observation group and control group, respectively, at baseline, and was (7.28±1.28)％ and (7.36±1.02)％ after 12 weeks of treatment [(-0.66±1.38)％ vs.(-0.64±1.14)％,P=0.914 5].The secondary outcomes were as follows: the fasting blood glucose (FBG) in the observation group were (7.91±1.87)mmol/L and (8.05±2.33)mmol/L at baseline and after 12 weeks of treatment;while in the control group, the FBG were (8.51±1.68)mmol/L and (8.00±2.02)mmol/L, and comparisons between two groups showed no significant change (P>0.05).The glycated albumin in the observation group and control group were (21.38±5.74)％ and (21.93±6.51)％ at baseline;after 4 weeks of treatment, they were (20.08±6.05)％ and (20.58±7.30)％ (the changes from baseline in these two groups were (-1.19±4.37)％ and (-1.20±5.08)％];after 8 weeks of treatment, they were (19.07±5.56)％ and (20.83±8.74)％ [the changes from baseline were (-2.09±4.51)％ and (-0.98±6.85)％];after 12 weeks of treatment, they were (19.03±5.19)％ and (19.36±6.14)％ [the changes from baseline were (-2.18±4.60)％ and (-2.47±5.20)％], there were no significant differences between two groups (P>0.05).The subgroup analysis showed that in those patients with the characteristics including body mass index (BMI)≥25 kg/m2 at baseline, the duration of diabetes mellitus longer than 8 years, fasting blood glucose less than 8 mmol/L and using insulin at baseline, the changes of HbA1c from baseline to the end of 12 weeks therapy in the observation group were more than in the control group.There were no significant differences between the two groups in terms of safety profiles, including the vital signs and laboratory findings (blood cell counts, liver function, and kidney function, all P>0.05).Conclusion Administration of BTVBT in T2DM patients for 12 weeks does not remarkably improve the HbA1c.

Objective To induce the differentiation of human umbilical cord blood MSCs into osteoblasts in vitro,and to study the method of inducing MSCs to differentiate into osteoblasts under specific microenvironment.Methods MSCs was obtained from human umbilical vein,and isolated by density gradient method.The morphological changes of MSCs were observed by using dexamethasone,beta sodium phosphate and vitamin C.The proliferation and differentiation of MSC in cord blood were studied by means of optical microscope,transmission electron microscope and alkaline phosphatase staining.The expression of bone morphogenetic protein 2 mRNA in human umbilical cord blood MSCs after osteogenic was inducted by RT-PCR.Results After the umbilical cord blood MSCs were differentiated into osteoblastic cells in microenvironment,fusiform cells became polygonal,irregular shape,local cells presented overlapping growth.After 10 days,the cells gradually presented square,crystal particles of high refraction,and began to show the characteristics of osteoblasts.The expression of bone morphogenetic protein 2 mRNA was positive in alkaline phosphatase staining and alizarin red staining.Conclusion Human umbilical cord blood MSCs can be induced into osteoblasts in vitro,which is an ideal seed cell for bone tissue engineering.

Objective: To investigate the clinical characteristics, treatment and curative effect of primary hepatic angiosarcoma. Methods: A retrospective study was conducted on 10 patients with primary hepatic angiosarcoma who were treated in the Department of Hepatobiliary Surgery, the First Affiliated Hospital of Zhengzhou University from April 2012 to July 2018. There were 6 males and 4 females, with an average age of 55.1 (range 39.0 to 71.0) years. The clinical features, imaging findings, treatment methods and survival time of the patients were analyzed. Results: The clinical manifestations of 10 patients with primary hepatic angiosarcoma were non-specific and all were confirmed by pathology. Three cases had diffuse liver echo changes on ultrasound, and 7 cases had hypoechoic lesions with blurred borders. Space occupying lesions were found in 10 cases by CT. A total of five patients underwent partial hepatectomy, 1 case received chemotherapy, 1 case received transcatheter arterial chemoembolization (TACE), 2 case received molecular targeted therapy, and 1 case was untreated. From diagnosis to the last follow-up, the survival time of the 5 patients receiving surgical resection were 9 months, 15 months, 19 months, 21 months and 26 months. The survival time of the patient receiving chemotherapy was 7 months. The survival time of the patient receiving TACE was 3 months. Two patients were targeted for treatment, one survived for 32 months, another had been targeted for 15 months, and treatment was continuing. One patient survived for 1 month without treatment. Conclusions Primary hepatic angiosarcoma has a high degree of malignancy and a poor prognosis. No specific clinical manifestations and imaging findings, confirmed by pathological examination. Patients with partial hepatectomy have a better survival. Targeted therapy may be the most effective treatment for primary hepatic angiosarcoma in the future.

Objective The incidence of gastrointestinal symptoms in diabetes is higher than that of non-diabetes.Thus,the aim of the present study was to observe the efficacy and safety of bifidobacterium tetragenous viable bacteria tablets in the treatment of constipation in patients with type 2 diabetes mellitus.Methods This is a multicenter,randomized,double-blind,placebo-controlled,parallel group-comparison clinical research.The subjects were randomly divided into study group and control group according to 1 ∶ 1 ratio by computer generated random number method.The subjects were either treated with bifidobacterium tetragenous viable bacteria tablets (study group) or placebo (control group) for eight weeks,and they were followed up for four weeks without changing foundation therapy for diabetes.The primary outcome was the change of complete spontaneous bowel movements (CSBMs).Results A total of 234 subjects (the study group:116 cases;the control group:118 cases) from 7 centers were included in the present study.The baseline characteristics were comparable between the two groups.In the study group,the CSBMs at 0,2,4,8 and 12 weeks were 0.0(0.0,1.0),1.0(0.5,2.0),2.0(1.0,3.0),3.0(2.0,3.5),2.0(1.0,3.0) times per week,respectively,while the CSBMs of the control group at each corresponding weeks were 0.0(0.0,1.0),1.0(0.0,1.5),1.0(0.0,1.5),1.0(0.0,2.0),1.0(0.0,1.5)times per week,respectively.There is significant difference in CSBMs between the two groups (P＜0.05).Moreover,after 12 weeks treatment,the CSBMs over spontaneous bowel movements (SBMs) ratio in the study group was higher than that in the control group [0.53 (0.40,0.67) vs 0.33 (0.00,0.50),P=0.048],indicating a more complete evacuation sensation in the study group.More subjects in the study group (66.38％) reached Bristol stool classification of normal criteria than those in the control group (48.31％,P=0.005).There were significantly improvement of bowel function index in the study group [study group 42.7 (33.3,56.7),control group 60.6 (51.7,75.7),P＜0.000 1].Furthermore,the symptoms of constipation was improved,and the satisfaction for the treatment was high in the study group.There were no significant differences of the safety indicators between the two groups.Conclusions Bifidobacterium tetragenous viable bacteria tablets can be used in patients with type 2 diabetes mellitus and constipation.Compared with placebo,it improves constipation and has no obvious adverse effects.