Aim To investigate the effect of TaorenHonghua drug pair on intervertebral disc degeneration (IVDD) in rats.Methods Fifty healthy Wistar rats were randomly divided into control group,model group,sham group,meloxicam group and Taoren-Honghua drug pair group,with 10 rats in each group.We established dynamic and static forces imbalance of cervical disc degeneration model or sham surgery in rats.12 weeks later,rats were intragastrically administered with meloxicam,Taoren-Honghua drug pair or saline for 30 days.C4/5 and C6/7 discs were harvested from rats.ABOG staining was used for observation of intervertebral disc morphology,real time PCR for mRNA expressions of type Ⅱ collagen (Col Ⅱ) and type Ⅹ collagen (Col Ⅹ),and immunohistochemical staining for Col Ⅱ and Col Ⅹ.Results Compared with model group,Col Ⅱ expression increased,while Col X expression decreased in chondrocyte of intervertebral disc in Taoren-Honghua-treated group(P ＜ 0.01).Conclusion Taoren-Honghua drug pair could delay the degeneration of cartilage endplate in rat intervertebral disc.

OBJECTIVETo study the effect of Charcot-Marie-Tooth 2L disease causing gene K141N mutation in heat shock protein B8 gene (HSPB8) on cell viability.

METHODSBy using liposome transfection technique, (wt)HSPB8, (K141N)HSPB8 eukaryotic expression vector and green fluorescent protein (GFP) vector were transfected into SHSY-5Y cell, respectively. Twenty-four hours later, the cells were treated with 44 degree centigrade lethal heat shock for 40 minutes. The relative viability of SHSY-5Y cells in each group was tested by using tetrazole blue colorimetric method (methyl thiazolyl tetrazolium, MTT).

RESULTSThere were significant differences among the light absorption value of GFP, pEGFP-(wt)HSPB8 and pEGFP-(K141N)HSPB8 transfected groups after heat shock (P<0.05), indicating that the relative viability of cells overexpressed with (wt)HSPB8 and (K141N)HSPB8 was different from that of control cells. The viability of cells overexpressing (wt)HSPB8 was highest, followed by cells overexpressed with (K141N)HSPB8. The viability of cells tranfected with GFP only was the lowest.

CONCLUSIONHSPB8 may play an important role in the protection of cells under lethal heat shock treatment, and the K141N mutation can impair the protective effect.

Cell Line, Tumor ; Cell Survival ; genetics ; Charcot-Marie-Tooth Disease ; genetics ; metabolism ; Gene Expression Regulation ; Genetic Vectors ; genetics ; Heat-Shock Proteins ; genetics ; metabolism ; Humans ; Mutation ; genetics ; Protein-Serine-Threonine Kinases ; genetics ; metabolism

Objective · To investigate the impacts of hepatitis B virus (HBV) infection on the metabolomic phenotype of HepG2 human hepatoma cells.Methods · With gas chromatography-mass spectrometry (GC-MS), metabolite composition of HepG2 and HepG2.2.15 cells (derived from HepG2 cells transfected with a plasmid containing HBV) were analysed. Results · GC-MS analysis mainly found 34 metabolites in both HepG2 and HepG2.2.15 cells,including glycine (Gly), alanine (Ala), valine (Val), leucine (Leu), isoleucine (Ile), proline (Pro), serine (Ser), threonine (Thr), methionine (Met), cysteine (Cys), cystine, aspartic acid (Asp), glutamic acid (Glu), pyroglutamic acid, phenylalanine (Phe), tyrosine (Tyr), tryptophan (Trp), hypoxanthine, uracil,myo-inositol, lactic acid, succinic acid, linoleic acid, linolenic acid, palmitic acid, stearic acid, urea, cholesterol, etc. These metabolites were involved in multiple metabolic pathways including glycolysis and metabolism of fatty acids, amino acids, purines and pyrimidines. Compared with HepG2 cells,HepG2.2.15 cells had significantly higher levels in lactic acid, linolenic acid, Ala and Cys, but lower levels in Leu, Ile, Val, Phe, Met, Trp, Pro, Tyr, myoinositol and uracil. Conclusion · HBV infection dysregulates the metabolism of amino acids and fatty acids in hepatocytes. GC-MS analysis provides complimentary information about HBV-induced metabolic changes of host cells.

Objective To analyze the incidence and death of malignant tumors in Haishu District during 2011 to 2016, and to provide reference for the formulation and adjustment of prevention and control strategies and measures. Methods From the chronic disease monitoring system of Zhejiang Province, the data of the death report of residents in Haishu district and the monitoring report of chronic disease were obtained for 2011 to 2016 years. The mortality of malignant tumor, incidence of malignant tumor, and survival rate of malignant tumor in residents were statistically analyzed. Results The mortality of malignant tumor in Haishu district from 2011 to 2016 was 182.00/100, 000 to 215.43/100, 000, however the malignant tumor mortality rate of males were significantly higher than females (P<0.01) . Malignant tumor was the first death cause of residents in Haishu district, and the death rate of malignant tumor accounted for over 30％ of all deaths. The average life expectancy loss of residents was 4.36 years due to cancer. The incidence of malignant tumor increased with age (P<0.01), and males were significantly higher than females in 2011 and 2012 (P<0.05), however there was no significantly difference between male and female after 2012 (P>0.05) . The top five deaths of malignant tumors were lung cancer, gastric cancer, colorectal cancer, liver cancer and pancreatic cancer, and the incidence of these five malignant tumors from high to low was lung cancer, colorectal cancer, gastric cancer, liver cancer and pancreatic cancer. The five year survival rate from high to low was colorectal cancer, gastric cancer, lung cancer, liver cancer and pancreatic cancer, respectively. Conclusion Malignant tumor has become the main cause of threat to the life and health of residents in Haishu district. It is suggested to strengthen the screening of early diagnosis and treatment of cancer.

OBJECTIVETo explore clinical efficacy of closed reduction and external fixation under local anesthesia for the treatment of high-risk elderly patients with intertrochanteric fracture.

METHODSFrom March 2013 to March 2015, 10 patients with intertrochanteric fractures treated with closing reduction and external fixator under local anesthesia were analyszed, including 4 males and 6 females, aged from 69 to 88 years old with an average of 75.2 years old. All fractures were caused by injury and classified to type I (5 cases), II (3 cases), and V (2 cases) according to Evans classification. According to American Society of Anesthesiologists (ASA), 6 cases were type III and 4 cases were type IV. Blood loss,operative time,hospital stays, postoperative complications, ambulation time and fracture healing time were observed, and Harris scoring were used to evaluate hip joint function.

RESULTSAll patients were followed up from 3 to 23 months with an average of 13.1 months. One patient with chronic obstructive pulmonary disease died for non-operation reason at 4 months after operation, the other fractures were healed at stage I, the mean fracture healing time was 5.6 months. There were no coxa vara, lower limb venous thrombosis, loosen and remove of needle passage. The average operative time was 46 min, blood loss was (35.00 ± 8.46) ml without blood transfusion. One patient was occurred pulmonary infection and stent-tract infection on the 2 nd and 3 rd day after operation, and improved with active anti-infection and dressing change; the other patients gone to ground activity at 4.2 d after operation. The patients stayed hospital for 10.6 d on average. According to Harris scoring at final following-up, the total score was 83.42 ± 3.27, 3 cases obtained excellent results, 5 cases good and 1 case poor.

CONCLUSIONClosed reduction and external fixation under local anesthesia in treating high-risk elderly patients with intertrochanteric fracture,which has advantages of shorter operative time, less blood loss, good recovery of postoperative function, is a safe, stable and economic method.

Aged ; Aged, 80 and over ; Anesthesia, Local ; Bone Nails ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; Fracture Fixation, Intramedullary ; Fractures, Closed ; surgery ; Hip Fractures ; surgery ; Humans ; Male ; Treatment Outcome

Objective To evaluate and compare the adult neural stem cells (NSCs) derived from the subventricular zone (SVZ), adipose tissue (AD) and bone marrow (BM) in SD rat in terms of their morphologies, their potential of neural differentiation and their ability to secrete neurotrophins (NTs).Methods Tissues from the suventricular zone, adipose tissue and bone marrow in the same SD rat were chosen and induced in vitro into SVZ-NSCs, AD-NSCs and BM-NSCs, respectively. The abilities of proliferation and differentiation of these 3 NSCs were compared. Immunocytochemistry and Western blotting were employed to detect the expressions of surface markers of neurons and neuroglia, including nestin,βtubulin, galactocerebroside C (GalC) and glial fibrillary acidic protein (GFAP). The secretions of BDNF and NGF were detected by ELIS A. Results No obvious differences of morphology between SVZ-NSCs and both BM-NSCs and AD-NSCs were found (P>0.05). The proliferation ability of SVZ-NSCs was stronger than that of BM-NSCs and AD-NSCs. The percentage of nestin-positive cells in the SVZ-NSCs was significantly higher than that in the BM-NSCs or AD-NSCs (P<0.05). No obvious differences in the expressions of βtubulin, GFAP, and GalC among the 3 groups were found (P>0.05).The secretions of BDNF and NGF in all the 3 groups could be noted; those in the SVZ-NSCs was significantly higher than those in the BM-NSCs and AD-NSCs (P<0.05); those in the AD-NSCs was slightly higher than those in the BM-NSCs. Conclusions SVZ-NSCs, AD-NSCs and BM-NSCs show similar morphological and phenotypic characteristics; however, SVZ-NSCs present more powerful proliferation, differentiation and secretion abilities than AD-NSCs and BM-NSCs. Considering about such problems as immuno-repulsion, ethic and the origins, AD-NSCs appear to be the better choice than BSVZ-NSCs and M-NSCs.

Objective To investigate the effect of collagen scaffold loaded with collagen-binding domain neurotrophin-3 (CBD-NT3) on the extension of cellular processes of dorsal root ganglions (DRGs), and explore the significance of this kind of combinatorial strategies in the spinal cord injury repair. Methods The tail tendons of SD neonatal rats were performed the removal of cellular components to prepare the collagen scaffold; HE staining was employed to evaluate whether the cells were completely removed from the collagen scaffold. The collagen scaffold was loaded with CBD-NT3,and then, they were co-cultured with primary DRGs for 1, 3 and 5 d, respectively. NT3 and PBS were also co-cultured with primary DRGs for 1, 3 and 5 d, respectively, as controls. Cells on the scaffold were stained by fluorescein diacetate (FDA) for morphology observation and the lengths and angles of the processes in each group were also quantitatively analyzed. Scanning electron microcopy (SEM) was employed to observe the topography of scaffold and the ultrastructure of DRGs 3 d after the co-culture.Results HE staining indicated that the cellular components in the scaffold were removed completely.The length of processes elongation in CBD-NT3 treatment group was significantly longer than that in the controls 3 d after the co-culture (P＜0.05). The 95% confidence interval of the angle between the line which the process emerged from the cell soma to the growing tip of the process and the long axis of fiber was 18.8-20.7 degrees. The results of SEM showed that cells could rely on the topography of the scaffold to anchor and grow. Conclusion The combinatorial strategies of collagen scaffold with CBD-NT3 can play a double function for oriented guiding and inducing extension of cellular processes effectively,which may provide a better therapeutic approach for spinal cord injury repair.

OBJECTIVETo clone the disease-causing genes possibly existing in 6.8 cM distance between microsatellite markers D12S1720 and D12S1611 in chromosome 12q24 for Charcot-Marie-Tooth disease type 2L (CMT2L).

METHODSTen positional and functional candidate genes were chosen among all known genes in this locus region by bioinformatics inqury. Mutation detection was performed by sequencing the exons and intron-exon junctions of the candidate genes.

RESULTSEleven sequence variations, that included 5 heterozygous and 6 homozygous variations, were detected in the exons and flanking areas of the 10 candidate genes. All the variations showed no co-segregation with disease phenotype.

CONCLUSIONTen candidate genes(TAOK3, RAB35, RPLP0, PXN, RNF10, RHOF, VPS33A, RSN, DENR, RNP24) were ruled out as the disease-causing gene for CMT2L. Ten single nucleotide polymorphisms (SNP) were reported for the first time.

Base Sequence ; Charcot-Marie-Tooth Disease ; genetics ; Chromosomes, Human, Pair 12 ; genetics ; Cloning, Organism ; DNA ; analysis ; DNA Mutational Analysis ; Humans ; Molecular Sequence Data ; Nucleic Acid Amplification Techniques

OBJECTIVETo evaluate left ventricular (LV) function and twist in patients with diabetic cardiovascular autonomic neuropathy (CAN) by two-dimensional speckle tracking imaging (STI).

METHODSSTI was performed in 56 subjects with type 2 diabetes mellitus (DM) (35 with DM only: group A, 21 with CAN: group B) and 34 normal subjects (Control) from LV short-axis view. LV peak systolic, peak early (E') and peak late (A') diastolic circumferential strain in 18 myocardial segments were measured at the levels of mitral annulus, papillary muscle and apex and the rotation at mitral annulus and apex levels were also measured. LV peak systolic and the ratio of E' and A' of global and three levels, twist, untwisting rate and untwisting half-time were calculated.

RESULTSIn group A, compared with control group, LV peak systolic radial circumferential strain has no significant difference (P > 0.05), E'/A' was reduced (P < 0.05), twist at aortic valve closure and twist at mitral valve opening were significantly increased (P < 0.05), untwisting rate reduced, and untwisting half time delayed. In group B, compared with control group and group A, circumferential strain parameters [(-12.64 ± 6.49)% vs. (-19.11 ± 9.98)% and (-21.14 ± 10.13)%, P < 0.05] and E'/A' [(0.90 ± 0.35) vs. (1.24 ± 0.47) and (1.98 ± 0.63), P < 0.05] were significantly decreased, twist at aortic valve closure [(19.08 ± 5.62)° vs. (16.57 ± 2.84)° and (14.36 ± 4.06)°, P < 0.05] and twist at mitral valve opening [(13.99 ± 2.31)° vs. (11.36 ± 2.63)° and (9.04 ± 5.63)°, P < 0.05] were significantly increased, untwisting rate [(0.40 ± 0.28)%/ms vs. (0.46 ± 0.14)%/ms and (0.53 ± 0.21)%/ms, P < 0.05] reduced, and untwisting half time [(489.61 ± 97.14) ms vs. (445.21 ± 54.53) ms and (410.60 ± 50.23) ms, P < 0.05] delayed.

CONCLUSIONSpeckle tracking imaging could be used to evaluate early changes on LV twist deformation and LV systolic function in patients with type 2 diabetes mellitus.

Diabetes Mellitus, Type 2 ; diagnosis ; physiopathology ; Diabetic Neuropathies ; diagnosis ; physiopathology ; Diagnostic Imaging ; methods ; Diastole ; Female ; Humans ; Male ; Middle Aged ; Rotation ; Stroke Volume ; Systole ; Ventricular Dysfunction, Left ; physiopathology ; Ventricular Function, Left

OBJECTIVETo study the possible mechanism of the intracellular aggregate formation of small heat shock protein HSPB8 (HSPB8)(K141N) mutation resulting in axonal Charcot-Marie-Tooth disease type 2L(CMT2L).

METHODSThe cell models which transiently expressed pEGFPN1-HSPB8 and pEGFPN1-(K141N)HSPB8 were established. The immunofluorescent co-location study of EGFP-(K141N)HSPB8 and HSPB1, EGFP-(K141N)HSPB8 and neurofilament light chain (NEFL) was carried out in the SHSY5Y cell models. The aggregate formation of EGFP-(K141N)HSPB8 in cell models was investigated and the possible mechanism of cellular aggregate formation was analyzed by t test and analysis of variance between group(ANOVA).

RESULTSEGFP-(K141N)HSPB8 formed large aggregate which predominantly located around the nucleus in cell models. EGFP-(K141N)HSPB8 co-localized perfectly with HSPB1 and NEFL in the SHSY5Y cell models. The aggregate formation was different in different cell types, there were fewer aggregates formed in an sHSPs deficient milieu than in HEK293T cells.

CONCLUSION(K141N)HSPB8 formed aggregates predominantly locate around the nucleus in cells. (K141N)HSPB8 co-localizes perfectly with HSPB1 and NEFL. The aggregate formation may be due to (K141N)HSPB8 conformational change leading to self aggregation and its abnormal interaction with other sHSPs such as HSPB1.

Cell Line ; Cell Line, Tumor ; Cell Nucleus ; metabolism ; Charcot-Marie-Tooth Disease ; genetics ; metabolism ; Green Fluorescent Proteins ; genetics ; metabolism ; HSP27 Heat-Shock Proteins ; HeLa Cells ; Heat-Shock Proteins ; genetics ; metabolism ; Humans ; Kidney ; cytology ; metabolism ; Microscopy, Confocal ; Neoplasm Proteins ; genetics ; metabolism ; Neuroblastoma ; genetics ; metabolism ; pathology ; Neurofilament Proteins ; genetics ; metabolism ; Point Mutation ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection