Objective In cartilage tissue engineering,it was a difficult problem to keep chongdro-cytes phenotype for long time.The aim was made to investigate the dedifferentiation of human articular chon-drocytes,and to verify the inhibition to dedifferentiation by transfection with recombinant pcDNA3.1(+)/TGF-? 2 in vitro.Methods Free chondrocytes were isolated from healthy articular cartilage harvested from6adult patients underwent operation because of other condition.The recombinant pcDNA3.1(+)/TGF-? 2 con-structed in advance was transferred by liposome mediated transfection to monolayer cultured articular chon-drocytes.The first,sixth and ninth generation chondrocytes being transfected and non-transfected were as-sayed by histological examination of HE and Safranine-O staining to observe their morphological changes.RT-PCR,ELISA,immunohistochemistry analysis and prime hybridization were performed to these cells to i-dentify the expression and biosynthesis of TGF-? 2 and cartilage-associated genes and proteins.Results The non-transfected articular chondrocytes in monolayer culture demonstrated the tendency of dedifferentia-tion,and lost the characteristics of their phenotype progressively in subculture.The expression of TGF-? 2 ,collagen typeⅡand proteoglycan in non-transfected cells were decreased,but expression of collagen typeⅠincreased gradually with time going on.On the contrary,as for the transfected chondrocytes,the trans-ferred gene was expressed in cells of all examined generations,and the expression levels were higher than that of non-transfected cell of same generations.The transfected chondrocytes maintained their phenotype and characteristics of articular cartilage.They expressed and produced collagen typeⅡand proteoglycan in a higher level,and collagen typeⅠin a lower level than that of non-transfected cells.Conclusion Human articular chondrocytes in monolayer culture have the tendency of dedifferentiation.The results suggested that chondrocytes tranfected by pcDNA3.1(+)/TGF-? 2 could produce endogenous TGF-? 2 .Transfection of pcD-NA3.1(+)/TGF-? 2 is able to provide transient and long lasting expression of TGF-? 2 in chondrocytes,and may be a feasible method to inhibit dedifferentiation of articular chondrocytes in vitro.

Objective To explore the clinical effect of whole body vibration training on stroke patients with hemiplegia and its effect on patients'balance function.Methods Eighty-six stroke patients with hemiplegia admitted to the Second Hospital of Jiaxing from June 2017 to December 2018 were divided into control group (43 cases)and observation group (43 cases) by random number table method.The control group received routine training,while the observation group received whole body vibration training on the basis of the control group.Both two groups were treated for 12 weeks.The changes of upper limb motor function,daily living ability and Berg balance scale (BBS)scores before and after intervention were compared between the two groups.Results After intervention,the FMA scores of the control group and the observation group were (37.89 ± 5.34)points and (49.61 ± 5.47)points,respectively,which were higher than before intervention (observation group:t =21.706,control group:t =10.272,all P ＜0.05).The FMA score of the observation group was higher than that of the control group (t =10.054,P ＜0.05).The improved Barthel index (MBI) scores of the observation group and the control group were (68.98 ± 7.91)points and(52.15 ± 6.87) points,respectively,which were higher than those before intervention (observation group:t =22.170,control group:t =10.119,all P ＜ 0.05).The MBI score in the observation group was higher than that in the control group (t =10.534,P ＜ 0.05).The BBS scores of the observation group and the control group were (59.98 ±6.47) points and (48.98 ± 3.71) points,respectively,which were higher than those before intervention (observation group:t =20.820,control group:t =13.877,all P ＜0.05).The BBS score in the observation group was higher than that in the control group (t =9.672,P ＜ 0.05).Conclusion Whole body vibration training has good clinical effect on stroke patients with hemiplegia,and it can improve upper limb motor function,daily living ability and balance function.

BACKGROUND: Bone marrow mesenchymal stem cells(MSCs) have been investigated initially. Because both fat tissue and bone marrow are tissues originated from mesoderm, so whether MSCs could be obtained from fat tissue as well, which also have multi-lineage differentiation potential?OBJECTIVE:To investigate the basic biological characteristics of adipose MSC and its differentiation into osteoblast under given culture condition for the exploration of its feasibility as seed cell in bone tissue engineering.DESIGN: A randomized and controlled study based on cells.SETTING: Department of Orthopaedics of an Affiliated Hospital of a university.MATERIALS: The study was conducted in the Department of Orthopaedics of the Third Hospital of Peking University. Adipose MSC extracted from the fat tissue of Lewis rats was used as subject.METHODS: Adipose MSCs were obtained from inguinal fat pads of Lewis rat after digestion, which were induced into adipocytes and osteoblasts with adipose and osteogenesis induced culture mediums. The differentiations were examined with cytochemical staining, immuncytochemical staining and Western blotting.MAIN OUTCOME MEASURES: Morphological and biological characteristic of adipose MSCs, and the specific mark of osteoblast after induction.RESULTS: Adipose MSCs were obtained from rat adipose tissue culture,which appeared fibroblast-likely in the culture in vitro, and could stably proliferate and passage in vitro. Primary adipose MSCs could differentiate into adipocytes spontaneously, and passaged cells could form fat drop under the reaction of insulin and dexamethasone, and then differentiate towards adipocytes after the expression of peroxidase proliferation activated receptor ?(PPAR-?) enhanced. There was significant difference between induction group and control group in alkaline phosphatase(ALP) activity detection under the induction of dexamethasone, ascorbic acid, and ?-sodium glycerophosphate(P ＜ 0.01) . Calcium node appeared in yon Kossa staining. Result of osteopontin (OPN) immunocytochemcial staining was positive,and OPN expression was detected by Western blotting after induction.CONCLUSION: MSCs with multi-lineage potential can be obtained from rat adipose tissue, and differentiate into adipocytes and osteoblasts after inductions. Therefore, adipose MSCs can possibly be served as one of optimal seed cells for bone tissue engineering.

Objective To investigate the effects of exercising on a cycle ergometer on lower limb motor function and the walking ability of hemiplegic stroke patients.MethodsSixty hemiplegic stroke patients were randomly divided into a treatment group and a control group with 30 in each. Both groups were treated with routine rehabilitation, but the treatment group also undertook cycle ergometer training. The training lasted for 10-20 min/d, 6 d/week for 6 weeks. All patients were assessed with the Fugl-Meyer lower limb assessment ( FMA), functional ambulation category (FAC) classification and the modified Barthel index (MBI) pretreatment and 6 weeks post treatment.ResultsBefore intervention there was no significant difference between the two groups in any of the assessments. Six weeks post treatment, the average FMA, FCA classification and MBI results in the treatment group were significantly better than those pretreatment and better than those in the control group.ConclusionsCycling exercise combined with routine rehabilitation training can distinctly improve the motor function of the lower limbs and the walking ability of hemiplegic stroke patients.

OBJECTIVE: To observe the physiological and biochemical effects of intermittent fasting combined with hunger-resistant food on mice, and to evaluate the safety and beneficial effects of this regimen. METHODS: One hundred and forty-four adult ICR mice were divided into 4 groups: standard feed AL group (ad libitum intake of standard feed), hunger-resistant food AL group (ad libitum intake of hunger-resistant food), standard feed IF group (feeding standard feed and fasting on alternate days), and hunger-resistant food IF group (feeding hunger-resistant food and fasting on alternate days). The experiment lasted for 4-8 weeks and all mice drank water freely. The quality of life, body weight, fasting blood glucose, serum lipid, blood routine test, liver and kidney functions as well as the viscera indexes were examined. RESULTS: Compared to the standard feed AL group, the caloric taking and the increment of body-weight were reduced (P<0.01), and the viscera indexes of the liver and kidney were elevated (P<0.05) in the hunger-resistant food AL group and the hunger-resistant food IF group, the values of fasting blood glucose were reduced in standard feed IF group and hunger-resistant food IF group (P<0.01), the value of triglycerides was reduced in hunger-resistant food IF group (P<0.05), while the quality of life, blood routine test as well as the liver and kidney functions were not obviously affected in the hunger-resistant food AL group, standard feed IF group and hunger-resistant food IF group. CONCLUSION: The regimen of intermittent fasting combined with hunger-resistant food is safe and beneficial to metabolic regulation, such as controlling body-weight and adjusting blood glucose and serum lipid. It is expected that development of this regimen will be helpful to the control of obesity and diabetes, etc.

OBJECTIVE: To examine the effect of different reduced caloric intake on mice transplanted with S180 ascitic tumor. METHODS: The institute for cancer research (ICR) mice were randomly divided into control group, 3.0 standard feed (SF) group, 2.0 SF group and 1.3 SF group. The mice in control group were fed enough (about 5 g/d) dietary intake, while the amounts of dietary intake in the latter three groups were scaled down in the proportion of 65%, which were 3.0 g, 2.0 g and 1.3 g standard feed respectively. Meanwhile the essential vitamins were added to the latter three groups to keep the amount of intake the same as that of the control's. RESULTS: For most of the mice, the caloric intake obviously prolonged the mean survival days and improved the life quality was 7.14 kcal/d, and the fasting blood glucose level was 2-3 mmol/L. CONCLUSION: Properly reduced caloric intake and keeping lower blood glucose level is beneficial to prolonging the survival time of mice transplanted with S180 ascitic cancer.

Objective To examine the application of Montreal Cognitive Assessment (MoCA) in Parkinson' s disease (PD) patients with normal general cognitive function by Mini-Mental State Examination (MMSE) evaluation.Methods PD patients were examined with MMSE, and those having a normal ageand education-adjusted MMSE score were included in the further study of MoCA testing.The patients with MoCA score not less than 26 were selected into normal control PD-NC group, and the patients with less than 26 into cognitive impaired PD-CI group.Scores of MoCA subtests were used in PD-CI group and PD-NC group to characterize cognitive changes in PD patients with mild cognitive impairment (MCI).MoCA score in PD-CI group used as dependent variable, and sex, educational level, age, course of disease, Hamilton Depression Rating Scale (HAMD), Hamilton Anxiety Rating Scale (HAMA), Self-rating depression Scale (SDS), Self-rating Anxiety Scale (SAS) and Unified Parkinson' s Disease Rating Scale (UPDRS) were used as independent variable, the risk factors of CI in PD patients was analysed by Linear Regression Analysis.Results There are 52.6% (112/213) PD patients with MMSE ≥ 26 while their MoCA < 26.Significant differences were observed in subtests of MoCA in visuospatial, executive, naming, attention,language, abstract, delayed recall and orientation between PD-CI group and PD-NC group (all P <0.01).Univariate and multivariate regression analysis showed that educational level is the most significant factor in PD-CI (OR:0.72, 95% CI 0.64-0.81, P < 0.05).Conclusions There is a high proportion of PD patients whose MMSE test showed normal but MoCA test showed cognitive impairment.MoCA examination was used to detect cognitive function of PD patients.Furthermore we suggest consider the education level in PD patients when evaluate their cognitive function.

The clinical data of 13 patients diagnosed as Takayasu's arteritis with cardiomyopathy in Peking Union Hospital were reviewed.Echocardiography showed cardiomegaly (left chambers mainly),diffuse weakened myocardial wall movement and reduced left ventricular ejection fraction.Dilated cardiomyopathy was diagnosed in two patients.Glucocorticoid and cyclophosphamide were the essential medication for treatment.One patient died,1 patient was aggravated and the disease condition in remaining 11 patients was stable or improved.Takayasu's arteritis with cardiomyopathy is a rare disease and it attacks younger people.It is difficult to identify at onset,and the disease deteriorates rapidly.Echocardiography should be performed earlier to evaluate the heart function for early diagnosis and treatment.Administration of large dose of glucocorticoid and cyclophosphamide at early stage may improve the prognosis.

Objective To investigate the role of the 5-hydroxymethylcytosine (5-hmC) DNA modification in the onset of systemic lupus erythemosus (SLE),we compared tihe levels 5-hmC between SLE patients and normal controls.Methods With informed consent,whole blood was obtained from patients,and genomic DNA was extracted.Using hMeDIP-seq analysis and validation by quantitative real-time quantitative polymerase chain reaction (RT-PCR),we identified the differentially hydroxymethylated regions that were associated with SLE.Results There were 1 701 genes with significantly different 5-hmC levels at the promoter region in the SLE patients compared with the normal controls.The CpG islands of 3 826 genes showed significant difference at 5-hmC levels in SLE patients compared with the normal controls.Out of the differentially hydroxymethylated genes,three were selected for validation,including TREX1,CDKN1A,and CDKN1B.The hydroxymethylation levels of these three genes were confirmed by quantitative RT-PCR.Conclusion Our studies indicate that there are significant alterations of 5-hmC in SLE patients;these differentially hydroxymethylated genes may contribute to the pathogenesis of SLE.Such novel findings show the significance of 5-hmC as a potential biomarker or promising target for epigenetic-based SLE therapies.

Objective To compare clinical effect of gonadotropin releasing hormone agonist(GnRH-a) alone and GnRH-a combined with low-dose dydrogesteronea and estradiol valerate on sex hormone, hypoestrogenic symptoms, quality of life and bone mineral density (BMD)in treatment of endometriosis.Methods Seventy patients with moderate or severe endometriosis, who were diagnosed by laparotomy or laparoscopic surgery within two months, were randomly assigned into two groups.35 patients in GnRH-a group were treated by goserelin (3.6 mg)for three months, and 35 patients in add-back group were treated by goserelin (3.6 mg)combined with estradiol valerate 0.5 mg and dydrogesteronea 5 mg daily.Before and after the treatment, clinical parameters were recorded and analyzed, including visual analog scale (VAS), medical outcomes survey short form 36 (SF-36), Kupperman menopausal index(KMI), BMD, the serum level of follicle stimulating hormone (FSH), estradiol (E_2) and bone gla-protein (BGP) .The first menstruation and VAS were also followed up after treatment.Results Every 3 cases in two groups lost follow-up.(1)Reproductive hormone: the level of E_2 in add-back group [(94 ± 71) pmol/L]was significantly higher than (54±52) pmol/L in GnRH-a group(P <0.01).The level of FSH in add-back group [(3.0 ± 1.9) U/L]was significantly lower than (5.7 ± 2.9) U/L in GnRH-a group (P < 0.05).(2) VAS: after treatment, VAS in both group decreased significantly when compared with that before treatment(P < 0.05), and remained until menstruated.(3) KMI: KMI in add back-group (10 ± 8) was significantly lower than (14 ± 6) in GnRH-a group (P < 0.05).(4) BMD: compared with that before treatment, BMD decreased significantly after treatment in GnRH-a group (P < 0.05), no remarkable difference of BMD was observed before and after treatment in add-back group.Before treatment, serum BGP in both groups did not show statistical difference.After treatment, the level of BGP in GnRH-a group [(7932±5206) ng/L]was significantly higher than (5419±2917) ng/L in add-back group (P <0.05).Conclusions GnRH-a combined with estrogen-progesterone regimen could relieve pain from endometriosis as effectively as GnRH-a alone and reduce hypoestrogenic symptoms and bone loss.Therefore,it is a safe and effective treatment.