ObjectiveTo investigate the relationship between the degenerative mechanisms of lumbar intervertebral disc (LID) and apoptosis.MethodsThe total RNAs were isolated from human LID tissues. Both the mRNAs from the degeneration and normal LID were reversely transcribed to the cDNAs. The cDNAs were labeled with the incorporations of fluorescent dUTP, for preparing the hybridization probes. The mixed probes were then hybridized to the cDNA microarray. After high-stringent washing, the cDNA microarray was scanned for the fluorescent signals and analyzed by computer image analysis. The apoptotic status and the expression of Bcl-2 and Bax in 12 cases of degenerative LID and 10 cases of normal LID were detected with TdT-mediated dUTP-biotin nick end labeling (TUNEL) and immunohistochemistry methods.ResultsAmong the 4096 targets, there were 10 genes related to apoptosis. The expression related to Bax protein gene was up-regulated and it was down-regulated for Bcl-2 protein. In group of normal LID, the average apoptotic index (AI) was (24.897±3.620); percentage of Bcl-2 positive cells was (31.440±4.150)%; percentage of Bax positive cells was (29.372±2.588)%, average optical density (OD) values of positive particles were (0.183± 0.010 ), ( 0.203 ±0.012) and (0.169±0.005) respectively. In group of degenerative LID, the average AI was (49.232±3.440); percentage of Bcl-2 positive cells was (18.239±2.470)%; percentage of Bax positive cells was (52.349±3.764)%; average OD values of positive particles were (0.152±0.003), (0.310±0.008) and (0.262±0.014) respectively. There were significantly differences in AI and expressions of Bcl-2 and Bax proteins between normal LID and degenerative LID (P<0.05).ConclusionCell apoptosis plays an important role in the process of LID degeneration. Both Bcl-2 and Bax take part in the occurrence and progression of LID.

Objective:To investigate the single nucleotide polymorphisms(SNP), haplotypes and genotypes of mannan-binding lectin(MBL) gene in the Bai(Pai) nationality from YunNan province, China.Methods:The three SNP sites CGT52TGT, GGC54GAC and GGA57GAA(named alleles D, B and C respectively, wildtype named A) in exon1 of MBL gene of 70 DNA samples of Bai nationality whose three SNP sites, -550G/C, -221C/G and +4C/T(named alleles H/L, X/Y and P/Q respectively), in promoter region of MBL gene had been clear, haplotypes and genotypes of MBL genes were detected and analyzed by sequence specific primer-polymerase chain reaction.Results:It was found that in Bais population, the frequency of alleles B was 0.100, there only were five haplotypes, HYPA, LXPA, LYQA, LYPA and LYPB, whose frequencies were 0.250, 0.107, 0.407, 0.135 and 0.100 respectively, the frequencies of several genotypes were LYPA/LYPA 0.043, LXPA/LYQA 0.143, LYPA/LYPB 0.014, HYPA/LYQA 0.086, LYPA/ LYQA 0.157, HYPA/LYPA 0.014, LYPB/LYQA 0.143, HYPA/LYPB 0.043, LXPA/LXPA 0.014, HYPA/LXPA 0.043, LYQA/LYQA 0.143 and HYPA/HYPA 0.157.Conclusion:In the MBL genes in Bais population, there is the allele B, the polymorphism haplotypes are mostly LYQA and HYPA, and the genotypes, LYPA/LYQA, HYPA/HYPA, LXPA/LYQA, LYPB/LYQA and LYQA/LYQA.

Resveratrol (trans-3, 4', 5-trihydroxy stilbene), a phytoalexin found in grape skins and red wine, has been reported to have a wide range of biological and pharmacological properties. It has been speculated that resveratrol may have cardioprotective activity. The objective of our study was to investigate the effects of resveratrol on intracellular calcium concentration ([Ca(2+)](i)) in rat ventricular myocytes. [Ca(2+)](i) was detected by laser scanning confocal microscopy. The results showed that resveratrol (15~60 mumol/L) reduced [Ca(2+)](i) in normal and Ca(2+)-free Tyrode's solution in a concentration-dependent manner. The effects of resveratrol on [Ca(2+)](i) in normal Tyrode's solution was partially inhibited by pretreatment with sodium orthovanadate (Na3VO4, 1.0 mmol/L, P<0.01), an inhibitor of protein tyrosine phosphatase, or L-type Ca(2+) channel agonist Bay K8644 (10 mumol/L, P<0.05), but could not be antagonized by NO synthase inhibitor L-NAME (1.0 mmol/L). Resveratrol also markedly inhibited the ryanodine-induced [Ca(2+)](i) increase in Ca(2+)-free Tyrode's solution (P<0.01). When Ca(2+) waves were produced by increasing extracellular Ca(2+) concentration from 1 to 10 mmol/L, resveratrol (60 mumol/L) could reduce the velocity and duration of propagating waves, and block the propagating waves of elevated [Ca(2+)](i). These results suggest that resveratrol may reduce the [Ca(2+)](i) in isolated rat ventricular myocytes. The inhibition of voltage-dependent Ca(2+) channel and tyrosine kinase, and alleviation of Ca(2+) release from sarcoplasmic reticulum (SR) are possibly involved in the effects of resveratrol on rat ventricular myocytes. These findings could help explain the protective activity of resveratrol against cardiovascular disease.
Animals ; Calcium ; metabolism ; Calcium Channels ; drug effects ; Heart Ventricles ; cytology ; metabolism ; Intracellular Fluid ; drug effects ; metabolism ; Male ; Myocytes, Cardiac ; drug effects ; metabolism ; Protein-Tyrosine Kinases ; drug effects ; Rats ; Rats, Sprague-Dawley ; Sarcoplasmic Reticulum ; drug effects ; metabolism ; Stilbenes ; pharmacology

OBJECTIVETo analyse retrospectively 3-tunnels core decompression with implantation of bone marrow stromal cells(bMSCs) and decalcified bone matrix (DBM) for treatment of femoral head necrosis in early stages, and to study its therapeutic results and indications.

METHODSAccording to the University of Pennsyvania system of classification and staging,to treat the patients with 3-tunnels core decompression with implantation of bone marrow stromal cells (bMSCs) and decalcified bone matrix (DBM) for treatment of early femoral head necrosis. Among the total 87 patients (103 hips), the male was 71 (86 hips) and the female 16 (17 hips) with the average age of 47 years old. The average course of disease was 1.3 year. The effect was evaluated by the clinical symptoms, the Harris' scores and the manifestations of radiology before and after operation.

RESULTSAll the patients were followed up more than 2 years (average 26 months). The average Harris' scores increased from preoperation 47.3 to postoperation 74.0. The average rate of excellent and good results was 75.7%. The rate of excellent and good was 88% (22/25) in type I, 78.7% (37/47) in type II and 61.3% (19/31) in type III. No severe complications were observed.

CONCLUSIONThree-tunnels core decompression with implantation of bMSCs and DBM not only removal the focus of disease, but also use DBM the induction of bone and autologous hone marrow stem cells to differentiate the functions of the femoral head can be resumed loading structure, eliminate pain,improve joint function. It is an effective method for early femoral head necrosis.

Adult ; Aged ; Bone Marrow Transplantation ; Bone Matrix ; transplantation ; Decompression, Surgical ; Female ; Femur Head Necrosis ; surgery ; Follow-Up Studies ; Humans ; Male ; Mesenchymal Stem Cell Transplantation ; Middle Aged ; Retrospective Studies ; Transplantation, Autologous

Objective To investigate the effect of polylactic-co-glycolic acid (PLGA)/graphene oxide (GO) nanofibers combined with brain derived neurotrophic factor (BDNF) on neural stem cells (NSCs) proliferation and differentiation as well as on the spinal cord injury repair.Methods PLGA/GO nanofibers were manufactured and absorbed with BDNF,and the microstructure of PLGA/GO nanofibers was observed by scanning electron microscope.The loading efficiency and release curve of BDNF on PLGA/GO nanofibers were measured by ELISA.NSCs were implanted on the surface of PLGA/GO and PLGA/GO/BDNF nanofibers.The absorbance values of each group were measured by MTT method,and the expression of Tuj-1 was observed by immunofluorescence and PCR.A total of 30 female SD rats were divided into control group (n =10),PLGA/GO group (n =10) and PLGA/GO/BDNF group (n =10) according to random number table.T9 spinal cord tissue was cut by Venus scissors to establish spinal cord hemisection injury model of rats.PLGA/GO and PLGA/GO/BDNF nanofibers were implanted onto the surface of injury site.BBB score was used to assess the motion functional recovery of the rats at 1,7,14 and 28 days after operation.Immunofluorescence staining of neuron specific nucleoprotein (NeuN)and glial fibrillary acidic protein (GFAP) were performed to observe the expressions of neurons and astrocytes at the injured site respectively one month after injury.Results The PLGA/GO nanofibers showed an irregular smooth fiber-like structure,and the average fiber diameter was (987.5 ± 176.3)nm.NSCs could differentiate into neurons on the nanofibers.The result of ELISA showed loading rate of BDNF on PLGA/GO nanofibers was about 47.5％.The release curve showed that BDNF was first released about 30％ on the first day and then about 60％ on the 21st day.The results of MTT and PCR showed that optical density value and Tuj-1 gene expression in the PLGA/GO/BDNF group were significantly higher than those in the PLGA/GO group (P ＜ 0.05).The animal experiment results showed that the BBB score of PLGA/GO/BDNF group was (15.3 ±0.7) points at 28 days after injury,which was significantly higher than that of the injury control group [(11.8 ± 0.8) points] and that of PLGA/GO group [(12.7 ±0.8) points] (P ＜ 0.05).Immunofluorescence results showed that the expression of NeuN in PLGA/GO/BDNF group was 13.7 ± 2.2,significantly higher than that in injury control group (4.3 ± 2.9) (P ＜0.05),and the expression of GFAP in PLGA/GO group was (25.6 ± 4.3) ％ significantly lower than that in injury control group [(38.5 ± 6.2) ％] and PLGA/GO group [(36.7 ± 7.3) ％] (P ＜ 0.05).Conclusion PLGMGO nanofibers combined with BDNF can effectively promote the proliferation and neuron differentiation of NSCs in vitro and repair spinal cord injury in vivo through orthotopic transplantation at the injury site.

To investigate the clinical features,treatment methods,and prognosis of a patient with hyperthyroidism crisis.The clinical characteristics,treatment methods,and prognosis of patients with hyperthyroidism crisis in our department were retrospectively analyzed.The application of adjuvant therapy and its influence on prognosis were summarized.Hyperthyroidism crisis is the most serious complication of those patients.The onset is urgent and the prognosis is poor,which requires the clinician to be vigilant and to interfere as early as possible.The extracorporeal circulation technique removes excessive free thyroxine,reduces the high metabolic state,and ensures safe of the heart.Brain,kidney,and other important organ functions which play important roles in the treatment on proper time.In this case,we finally achieved clinical cure,providing some new ideas for the diagnosis and treatment of hyperthyroidism crisis.

OBJECTIVETo construct Epithelia Membrane Protein 1 gene-deficient in human fetal nucleus pulposus model by lentivirus-mediated RNA interference for building a platform for illustrating the biomechanisms role of EMP-1 during human intervertebral disc degeneration.

METHODSThe lentivirus vector with shRNA targeting EMP-1 mRNA was transected into 293FT cells by liposome. Then the lentivirus supernatant was obtained and used for infecting human fetal nucleus pulposus. The expression of GFP was observed under fluorescence microscope after 48 h. The viral particles were collected at 72 h after transfection. The efficacy of gene interference was tested by Western blot and Real-time RT-PCR. Analysis the results of the fluorescent microscope scenes and get the average values of EMP-1/GAPDH by detected the interference efficiency of various interference DNA sequences with western blot and semi quantitative RT-PCR methods.

RESULTSThe lentivirns with high titer were obtained and the EMP-1 gene deficient cell strains were obtained. Semi quantitative RT-PCR and Western blot proved the average values of EMP-1/GAPDH decreased from 0.46 to 0.32 and 0.5 to 0.25 (P < 0.01).

CONCLUSIONLentivirus packaging technology can be mastered skillfully. EMP-1 gene-deficient cell models are successfully established.

Fetus ; HEK293 Cells ; Humans ; Intervertebral Disc ; metabolism ; Lentivirus ; genetics ; Neoplasm Proteins ; genetics ; RNA Interference ; Receptors, Cell Surface ; genetics ; Transfection

OBJECTIVETo clarify three-grade criteria of curative resection for primary liver cancer (PLC) and evaluate their clinical significance.

METHODSCriteria of curative resection of PLC were summed up to three grades. Grade I: complete removal of all gross tumors with no residual tumor at the excision margin. Grade II: on the basis of Grade I, there was no extrahepatic metastasis, no hilar lymph node metastasis, no tumor thrombus in the main trunks and their primary tributaries of the portal vein, common hepatic duct, hepatic vein and vena cava inferior, and the tumor was not more than two in number. Grade III: in addition to the above criteria, AFP dropped to normal level (in patients with elevated AFP before surgery) within 2 months after operation, and no residual tumor upon diagnostic imaging. A total of 354 cases with PLC who had their liver resected was reviewed. Patients in each grade were divided into two portions depending on whether the treatment was curative or palliative.

RESULTSThe survival of patients receiving curative treatment was better than those receiving palliative treatment (P < 0.01). This was true for patients whose treatment belonged to anyone of the three-grade criteria. The survival was improved along with the promotion of curative criteria used. The 5-year survival rate of Grade I, II and III patients undergone curative resection was 43.2%, 51.2% and 64.4%, respectively (P < 0.01).

CONCLUSION1. The three-grade criteria may be used for judging the radicality of tumor resection for PLC. 2. The more stringent the criteria used, the better the survival would be. 3. Adopting high-grade criteria to select cases, to guide operation and postoperative follow-up would improve the results of liver resection for PLC.

Female ; Hepatectomy ; methods ; Humans ; Liver Neoplasms ; mortality ; surgery ; Male ; Middle Aged ; Survival Rate

Adult ; Aged ; China ; ethnology ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; genetics ; Polymorphism, Genetic ; Thrombospondin 1 ; genetics

Objective To investigate the clinical characteristics of metabolically healthy obese ( MHO) individuals, and to explore the risk of progression into metabolic disorders after 3 years. Methods A total of 3943 residents in Jining City were evaluated twice from February 2012 to August 2015, and 3766 individuals were enrolled excluding those with missing data. Of the subjects, 875 subjects were screened as metabolic normal population, which were divided into MHO(n = 127), metabolically healthy overweight (MHOW, n = 386), and metabolically healthy normal weight ( MHNW, n = 362) groups. T test, x2 test, and logistic regression analysis were used for data analysis. Results The incidence of MHO was 11. 63％ (127 / 1092) in obesity, and the proportion of MHO in females was higher than that in males(13. 91％ vs 7. 82％ , P<0. 05). Compared with MHNW group, the levels of HbA1C , fasting insulin, low density lipoprotein-cholesterol ( LDL-C), triglyceride ( TG), glutamyl transpeptidase (GGT), systolic blood pressure(SBP), diastolic blood pressure(DBP), and waist circumference(WC) were higher in MHO while glomerular filtration rate (GFR) and high density lipoprotein-cholesterol (HDL-C) were lower(all P<0. 05); and fasting insulin, LDL-C, TG, GGT, SBP, WC were higher in MHOW while HDL-C was lower (all P<0. 05). The levels of fasting insulin, TG, SBP, WC were higher in MHO while GFR and HDL-C were lower compared with MHOW(all P<0. 05). Following up for 3 years, the incidences of dyslipidemia in MHNW, MHOW, and MHO were 17. 96％ (65 / 362), 32. 90％ (127 / 386), 42. 52％ (54 / 127), respectively, with significant difference among three groups(P<0. 05). The incidences of hyperglycemia in the three groups were 20. 17％ (73 / 362), 22. 80％(88 / 386), 26. 77％ (34 / 127), respectively, without significant difference among groups ( all P > 0. 05). After adjustment for some factors including sex, age, fasting insulin, glutamic pyruvic transaminase, glutamic oxaloacetic transaminase, GGT, and creatinine, the risks of dyslipidemia in MHO ( OR = 2. 193, 95％ CI 1. 359-3. 539, P<0. 05) and MHOW(OR= 1. 705, 95％ CI 1. 190-2. 443, P<0. 05) were significantly increased as compared with MHNW. Conclusion Compared with MHNW individuals, MHOW/ MHO individuals show an obviously different clinical feature as well as with higher risks of dyslipidemia after 3 years.