BACKGROUND: The functional gene fragments integrate into gene vector, which is then transfacted into target cells or joint cavity, through the transgenic target cells continue to secrete a large number of functional gene product, local therapeutic concentrations could be maintained within a long period of time, thus repairing articular cartilage injury. OBJECTIVE: To transfect rabbit articular chondrocytes using recombinant retroviral vector-mediated transforming growth factor-βl (TGFβ_1) in vitro, and to observe its expression and its effect on biological characters of chondrocytes. METHODS: Rabbit chondrocytas were isolated by use of trypsin digestion method. Vector was PLNCX_2 Hind Ⅲ/Not Ⅰ doubly digested and dephosphorylated, connected with some multiple cloning sites and RFP gene following pDsRed_2 double digestion, to build PLNCX_2-RFP. TGFβ_1 gene was amplified from the PGEMT-TGF and connected with PLNCX_2-RFP following double digestion, to build PLNCX_2-TGFβ_1-RFP. Subsequent to packaging retroviral vector, viral supernatant titer was detected. The cultured and transfected chondrocytes in rabbit knee joint were divided into 3 groups: control group (without any transfection), transfected PLNCX_2 group and transfected PLNCX_2-TGFβ_1-RFP group, continued screening 2 weeks to observe the cellular changes. Cell supematant transfected stably were collected for detecting the effect of gene transfection on the chondrocytes with NO detection kit, ELISA assay was applied to determine human TGFβ_1 expression in cell culture supernatant. RESULTS AND CONCLUSION: The recombinant gene PLNCX_2-TGFβ_1-RFP was identified correct sequence by the enzyme digestion sequencing TGFβ_1 and RFP, which showed that the eukaryotic expression vector PLNCX_2-TGFβ_1-RFP had been successfully built as expectation. They were then transfected into packaging calls and cultured, the virus titer was defined as 1×10~6 CFU. Following stable transfection of cartilage cells, red fluorescence can be observed, proving successful transfection. After continuous screening 2 weeks, the scattered adherent calls formed positive clones, and gradually diffusely integrated, cell clusters appeared with common dual cores, the calls proliferated actively. NO concentration in the transfected PLNCX_2-TGFβ_1-RFP group was higher than that of transfected PLNCX_2 group (P < 0.05), no difference was significant between control group and transfected PLNCX_2 group. The control group and the group transfected PLNCX_2 showed no TGFβ_1 expression, while TGFβ_1 concentration was (28.08±3.73) ng/L in the transfected PLNCX_2-TGFβ_1-RFP group. PLNCX_2 ratroviral vector-mediated human TGFβ_1 can be effectively transfected into rabbit knee joint cartilage cells and obtain stable expression, while the transfected cartilage calls proliferate actively.

AlM:To investigate biological measurement parameters of anterior segment in acute angle- closure glaucoma ( AACG) .METHODS:Forty-six eyes of 46 patients with AACG and 52 eyes of 52 patients with shallow anterior chamber and 50 eyes of 50 normal individuals were examined. The parameters of anterior segment including chamber crowd rate (CCR), lens thickness (LT), lens position (LP) and anterior chamber depth ( ACD ) were measured by A-ultrasound according to different ages in each group. The data were performed statistical analysis in three groups.RESULTS: ln each age range group (≥50 ~ 59 years old, ≥60 ~ 69 years old, ≥70 years old ) , statistically significant differences in three groups ( AACG, shallow anterior chamber group and the controls ) were found in CCR, LT, LP, ACD (P<0. 01). There were statistically significant differences only in CCR (P<0. 01) but not in LT, LP and ACD ( P > 0. 05 ) between AACG and shallow anterior chamber group in each age range group.CONCLUSlON: CCR can be used as the index of evaluating crowding state of anterior segment in AACG patients and the sensitivity is better than LT and LP.

Objective To observe the effects of astragaloside Ⅳ on electrocardiogram (ECG) and action potential of ventricular myocytes in guinea pigs. Methods ECG was recorded in vivo and ex vivo by using conventional ECG recording method from anesthetic guinea pigs and Langendoff perfusion model of hearts. Action potentials were recorded from isolated papillary muscles of right ventricles of guinea pigs by using microelectrode techniques. Results RR interval was prolonged by Astragaloside Ⅳ in a dose-dependent manner both in vivo and ex vivo. Astragaloside Ⅳ shortened action potential duration (APD), while had no effects on resting potential, action potential amplitude and maximal rate of depolarization. Conclusion Astragaloside Ⅳ exerts a negative chronotropic effect on heart and shortens APD of cardiac myocytes, which may be involved with calcium channels.

Objective To study the effects of sample digestion conditions on measurement results of urinary iodine determined by As(Ⅲ)-Ce4+ catalytic spectrophotometry with ammonium persulfate digestion,and to promote the application of newly revised (the 2012 edition) national standard method for determination of urinary iodine.Methods According to the newly revised national standard method,various digestion conditions,such as ammonium persulfate concentration (0.8-1.3 mol/L,group interval 0.1),digestion instruments (heating block and drying oven) and standing time after digestion(0.5,1.0,2.0,4.0 and 22.0 h),were studied.The samples included 3 standard materials,which were GWB09108k,GWB09109f and GWB09110m containing iodine of (68.2 ± 9.0),(138.0 ± 10.0) and (221.0 ± 10.0) μg/L,and 5 urine samples with iodine concentration of 100-300 μg/L.Results Measurement results among the three groups of 0.9,1.0 and 1.1 mol/L ammonium persulfate digestion fluid showed no significant difference(P ＞ 0.05).The digestive effect showed no significant difference between heating block and drying oven (P ＞ 0.05) except one standard material in low concentration (GBW09108k).After digestion,samples were placed 0.5-22.0 h,the measurement results between groups showed no significant difference (P ＞ 0.05).Conclusions Appropriate concentrations of ammonium persulfate are from 0.9 mol/L to 1.1 mol/L.Heating block is recommended for the digestion,however,when absent,drying oven can be used alternatively.The standing times from 0.5 h to 22 h after digestion have not affected the measurement results.

Chinese traditional medicine has provided, since ancient times, a basis for health care and medicine to the Chinese nation and for China's national stability. Identification of the constituents responsible for therapeutic and undesired effects of Chinese herbal medicines is a type of key research facilitating the modernization of these medicines. For a complex Chinese herbal medicine, multi-compound pharmacokinetic research is a useful approach to identifying its constituents that are bioavailable (in their unchanged and/or metabolized forms) at loci responsible for the medicine's therapeutic action and to characterizing the compounds' disposition and pharmacokinetics related to the action. In addition, such pharmacokinetic research is also useful for identifying herbal compounds associated with the medicine's adverse effects and drug-drug interaction potential. Over the past decade, great advances have been achieved in the theory, methodology, associated techniques, and their application of such multi-compound pharmacokinetic research, which has become an emerging field in pharmacokinetics. In this perspective, we elaborate on the methodology, technical requirements, and key analytical techniques of multi-compound pharmacokinetic research on Chinese herbal medicines, describe research examples regarding investigation of pharmacokinetics and disposition of a class of bioactive herbal constituents (ginsenosides of Panax notoginseng root) and pharmacokinetics-based identification of potential therapeutic compounds from a dosed Chinese herbal medicine (LianhuaQingwen capsule), and discuss follow-up development for the multi-compound pharmacokinetic research.

Objective To explore the clinical symptoms and neuroimaging features of a patient with atypical hypomyelinating leukodystrophy with atrophy of the basal ganglia and cerebellum (H-ABC) caused by a novel TUBB4A mutation.Methods We analyzed the clinical data, imaging features and the result of genetic testing of a case diagnosed as atypical H-ABC.Results The initial symptoms were progressive spasticity, mild cerebellar ataxia and mild cognitive impairment.MRI showed regional blurring of slight high signal on T2-weight and FLAIR image in white matter of the bilateral midbrain ventral, internal capsule, posteior horn of lateral ventricle and centrum semiovale, with normal bilateral cerebellar and caudoputamen nucleus.Compared with normal subjects of the same age and gender, hypometabolism was found by 18F-FDG-PET in brainstem, cerebellar and caudoputamen nucleus in the patient.Genetic testing revealed a de novo pathogenic exome missense heterozygous mutations c.70G>A in TUBB4A, which was not reported in the human gene mutation database (HGMDpro) and was assessed to be a pathogenic mutation by pathogenic mutation prediction software.Conclusions The diversity of TUBB4A gene mutations may cause different functional and/or structural impairment in subcortical white matter, cerebellar and caudoputamen nucleus, leading to atypical symptoms and neuroimaging features.Genetic testing for pathogenic mutation in TUBB4A gene is a key for the diagnosis of H-ABC.

0.05).(4) Significant differences between CC and CIN Ⅰ for p16,CDH1,RASSF1A and TIMP3 genes(P

Objective To analyze the characteristics of auditory verbal memory impairment in mild Alzheimer's Disease (AD) and Mild Cognitive Impairment (MCI).Methods Auditory verbal memory test was performed in 72 patients with MCI,45 patients with mild AD,and 62 normal controls.Results Significant intergroup differences were found in total former five free recall and learning scores,The MCI subjects( 16.4?5.5,2.6?1.7)performed significantly more poorly than the normal control subjects(NC) (30.2?5.6,3.4?1.9),and mild AD categories (9.8?4.1,2.0?1.2) showed lower results than the MCI subjects(t=2.26,P

Objective This study was to explore the prevalence of overweight and obesity,and the effects of contextual and individual level determinants on them in the rural areas of Kunming city.China.Methods Shilin County was selected as the study site.Probability Proportional to Size(PPS)sampling method was used to select representative sample of 6006 residents aged 45 years and over from Shilin.Information was obtained from a cross-sectional survey on health.Data was analyzed using a muhilevel logistic modeling.Results The prevalence rates of overweight and obesity were 12.10% and 2.15% in the study area.Males had a higher prevalence of overweight than females(13.60% vs.10.71%).Similar situation was seen in the prevalence of obesity(2.82% vs.1.52%).Both village level and individual level variables were associated with obesity.whereas only individual level variables were related to overweight.Elderly had lower probability of being overweight and obese than younger people with odds ratio(OR)as 0.95(95% CI:0.83-0.97)and 0.93(95% CI:0.82-0.96),respectively.Males had higher probability of being overweight and obese than females:OR of 0.89(95% CI:0.78-0.98)and OR of 0.87(95% CI:0.78-0.97),respectively.Individuals with lower family income had increased probability of having obesity (OR=0.81,95% CI:0.73-0.95).Factor as living in a higher income village was associated with lower prevalence of obesity(OR=0.92,95% CI:0.85-0.98).Conclusion Interventions at village level on obesity in parallel with those at individual level were needed.Prevention and intervention on obesity should be emphasized in villages with higher income.

ObjectiveTo probe the clinical features,diagnosis and treatment of idiopathic generalized myokymia.MethodsSeven patients with idiopathic generalized myokymia were analysed retrospectively.ResultsAll 7 patients showed prominent myokymia characterized by undulating and vermicular movements spreading across the muscle surface. The myokymia in gastrocnemius muscles in all cases. The myokymia also appeared in both upper extremities in 5 patients,and in faces,waist,back,abdomen and all extremities in 2 patients. Muscle rippling movement was induced and increased by exercise,and persistent during sleep. The vermicular myokymia could be observed easily in the relaxation of the muscles. Electromyography tests showed myokymic discharges in 5 patients,but normal in 2 patients. 5 patients of them were cured with carbamazepine and phenytoin sodium.ConclusionThere are typical clinical features and effective treatment in the patients with idiopathic generalized myokymia.