Objective To observe the effect and significance of high-expression HOXB4 in controlling proliferation and cycle of human cartilage endplate stem cells (CESCs).Methods CESCs were divided into adenovirus-mediated HOXB4 delivery group (Group A),empty virus delivery group (Group B) and blank control group.Gene and protein expressions of HOXB4 in Group A were detected by PCR and Western blot respectively; cell proliferation among those groups were determined using cell counting kit 8 (CCK8) technique; cell cycle among those groups was measured by propidium iodide (PI) assay and flow cytometry.Results (1) Over-expressed HOXB4 virus was transferred to CESCs successfully; (2) Real-time quantitative PCR results showed 3.6 times higher expression of HOXB4 in Group A than in blank control group.Western blotting indicated HOXB4 protein in Group A was 3 times the level in control group; (3) HOXB4 promoted CESCs proliferation (P ＜ 0.05) and blocked the cells at phase S.Cells at phase S in Group A was increased from 29.27 to 30.28 (P ＜ 0.05).Conclusion Over-expressed HOXB4 accelerates proliferation of CESCs and increases cell population at phase S,indicating that HOXB4 hindering CESCs degeneration may be an approach to treat lumbar intervertebral disc protrusion.

Central serous chorioretinopathy (CSC) is characterized by serous detachment of the sensory retina as a consequence of the focal leakage of fluid from the choriocapillaries to subretinal space through a defect of the retinal pigment epithelium(RPE). The exact cause of CSC is not well unknown. Psychological stress is thought to contribute to CSC, but the physiologic mechanisms are unclear. It is hypothesized that psychological stress can induce CSC through the mechanism of the hypothalamic-pituitary-adrenal (HPA) system. Psychological stress can adversely affect HPA axis and causes glucocorticoid levels to elevate. Increased glucocorticoids constrict choroid vessels, which leads to ischemia of choroids and damage vascular endothelial cells, thus causing vasopermeability to increase. RPE dysfunction will occur as a result of abnormalities in the choroidal circulation. The large molecules including protein may enter the subretinal space through the damaged vessels and RPE.

Adolescent ; Adult ; Aged ; Case-Control Studies ; Child ; Female ; Flow Cytometry ; Humans ; Male ; Middle Aged ; Platelet Glycoprotein GPIIb-IIIa Complex ; metabolism ; Purpura, Thrombocytopenic ; diagnosis ; metabolism ; Young Adult

OBJECTIVETo study the protective effect of Ligustrazine Injection (LI) against cisplatin-induced ototoxicity and to explore its mechanism.

METHODSThirty healthy adult guinea pigs were randomly divided into three groups, 10 in each group, i.e., the normal control group, the cisplatin group, and the LI group. Guinea pigs in the normal control group were intraperitoneally injected with normal saline at 3 mL/kg for 7 consecutive days. Those in the cisplatin group were intraperitoneally injected with cisplatin at 3 mg/kg for 7 consecutive days. Those in the LI group were intraperitoneally injected with LI at 140 mg/kg for 7 days, but cisplatin (3 mg/kg) was intraperitoneally injected from the opposite side starting from the 4th day. Brainstem auditory evoked potential (BAEP) was performed in all animals before and after injection. All animals were sacrificed after the final testing under anesthesia and their cochleas collected. Half the cochleas of each group were collected for silver nitrate staining of cochlear basilar membrane stretched. The other half the cochleas of each group made into paraffin sections to observe the apoptosis of cochlea cells by TUNEL method and the expression levels of c-Jun detected by immunohistochemistry.

RESULTSThere was no statistical difference in the difference of BAEP threshold among the 3 groups before injection (P > 0.05), but the BAEP threshold increased in the cisplatin group and the LI group (P < 0.05). Besides, it was higher in the cisplatin group (P < 0.05). In the cisplatin group, most hair cells were missing, spiral ganglion cells obviously decreased, more TUNEL positive cells occurred, and the expression of c-Jun was stronger. But the aforesaid impairment in the LI group was obviously lessened (P < 0.05).

CONCLUSIONSLI showed certain antagonist effect on cisplatin-induced ototoxicity. Its mechanism might be associated with scavenging oxygen radicals of the cochlea tissue, improving the microcirculation, and fighting against apoptosis.

Animals ; Apoptosis ; drug effects ; Cisplatin ; toxicity ; Cochlea ; drug effects ; metabolism ; pathology ; Evoked Potentials, Auditory, Brain Stem ; Guinea Pigs ; Pyrazines ; pharmacology ; Reactive Oxygen Species ; metabolism

OBJECTIVETo compare the early clinical effects of Activ C cervical disc replacement (ACDR) and anterior cervical discectomy and fusion (ACDF) in treating single-level cervical spondylosis.

METHODSThe clinical data of 76 patients with single-level cervical spondylosis underwent surgery from July 2009 to September 2012 were retrospectively analyzed. Among them, 28 patients were treated with ACDR (ACDR group), including 18 males and 10 females, aged from 32 to 62 years old with an average of (45.2±6.2) years; and 48 patients were treated with ACDF (ACDF group), including 28 males and 20 females, aged from 33 to 60 years old with an average of (45.8±6.4) years. Visual analogue scale (VAS), Japanese Orthopedics Association (JOA) score, Short Form-36 (SF-36), imaging data were used to assess the clinical effects after operation.

RESULTSA total of 76 patients were followed up from 6 to 24 months with an average of 13.2 months. VAS of neck pain and brachialgia were improved in all patients after operation (P<0.05), there was no significant difference between two group (P>0.05). Somato-score and psycho-score of SF-36 of two groups were obviously increased (P<0.05), ACDR group was better than that of ACDF group (P<0.05). In ACDR group, there was no significant difference in the range of motion of surgical segments and adjacent segments between preoperative and postoperative (P>0.05); heterotopic ossification around the edge of vertebral body occurred in 1 case on the 6th month after operation, no fusion was found on the 1st year after operation. In ACDF group, the adjacent vertebral disease occurred in 1 case and the patient underwent the reoperation.

CONCLUSIONActiv C cervical disc replacement can reduce the degeneration of adjacent segments and its early outcomes for the treatment of single-level cervical spondylosis are satisfactory, but the long-term effects still need study.

Adult ; Cervical Vertebrae ; surgery ; Diskectomy ; methods ; Female ; Humans ; Male ; Middle Aged ; Spinal Fusion ; methods ; Spondylosis ; surgery ; Total Disc Replacement ; methods

Objective To study the relation of matrix metalloproteinase-9 in serum and tumor necrosis fac- tor-?in serum and amniotic fluid in predicting ehorioamnhionitis in patients with premature rupture of membranes (PROM).Methods The levels of MMP-9 in serum and TNF-?in serum and amniotie fluid were measured by ra- dioimmunoassay and ELISA in 67 cases with premature rupture of membranes as study group and 40 cases normal full-termed pregnant women as controls group.Results(1)The levels of TNF-?in amniotie fluid and MMP-9 in serum in study group were significantly higher than those in controls group(P0.05).(2)In study group,the levels of MMP-9 of serum in0.05).Conclusions The levels of TNF-?in amniotic fluid and MMP-9 in serum were valuable clinical indices for identification of chorioamnionitis in patients with PROM.The levels of MMP-9 in serum also could assess the time of rupture of membranes and the degree of ehorioamnionitis.

Objective To describe the clinical characteristics of a patient with Gitelman syndrome,and to identify the associated SLC12A3 gene mutations.Methods A suspected case of teenager-onset Gitelman syndrome was observed in our hospital.It was further confirmed by clinical manifestations and auxiliary examination.In addition,direct sequencing for the exons of SLC12A3 gene and CLCNKB gene region was conducted to identify the probable disease-associated mutations.Results The case showed characteristics of hypokalemia,hypomagnesemia,and low level of urinary calcium and onset by age of 18.By excluding the possibilities of long-term use of thiazide diuretics,laxatives,chronic vomiting and diarrhea,he was finally diagnosed as a case of Gitelman syndrome.Furthermore,by Sanger direct sequencing,2 coding variations were identified in SLC12A3 gene region,including T304M and L488P.L488P was a new heterozygous mutation.Conclusion Detection of SLC12A3 gene mutation could facilitate the diagnosis of Gitelman syndrome and improve prognosis.

Five flavone C-glycosides were isolated from the methanol extract of the degrease seeds of Ziziphus jujuba var. spinosa though various column chromatography methods including silica gel, MPLC, and HPLC. The structures were elucidated as 6"-feruloyl- 6'''-vanillylspinosin(1), 6",6'"-diferuloylspinosin(2), spinosin(3), swertisin(4) and isoswertisin(5) based on the NMR and MS spectral data. 1 is a new compound.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Flavones ; chemistry ; isolation & purification ; Glycosides ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; Mass Spectrometry ; Molecular Structure ; Seeds ; chemistry ; Ziziphus ; chemistry

Objective To establish a specific fluorescence quantitative method for determining the mRNA expression of Toll-like receptor3(TLR3)in peripheral blood mononuclear cells(PBMCs).Methods Using the Beacon Designer 2.1 software,specific primers and Taqman-MGB probe were designed.The plasmid pMD18-T-TLR3 was constructed as calibrator and the amplified fragment was obtained by reverse- transcript-PCR(RT-PCR).RNA quantification based on cycle threshold values(Ct)was used to establish the standard curve.According to which,the TLR3 mRNA levels in 30 normal individuals,20 patients with primary biliary cirrhosis(PBC)and 20 ones with chronic liver cirrhosis induced by HBV were calculated automatically by software after the fluorescence of PCR product was detected continuously during amplification.Results The linear detection range of the assay for TLR3 gene and ?-actin was 10~2-10~8(r= -0.9974)and 10~3～10~8(r=-0.9984),respectively.The coefficient of variation of both intra-and inter- assay reproducibility for high concentration sample were 6.7% and 8.7%,respectively,and those for low concentration sample were 12.3% and 14.0%.The TLR3 mRNA expression level ranges from 3.46?10~2- 4.51?10~3 copies/?g RNA,4.92?10~2-1.42?10~4 copies/?g RNA and 2.58?10~2-7.17?10~3 copies/?g RNA for 30 healthy individuals,20 PBC patients and 20 ones with chronic liver cirrhosis induced by HBV, respectively.Conclusion We have successfully set up a FQ-RT-PCR method for detecting TLR3 mRNA, which may be used as an excellent tool for the clinic and basic study on the expression of TLR3 gene.

Objective:To construct a DNA vaccine co-expressing the HBV compound multi-epitope antigen gene, the hIL-12 and the anti-HBV siRNA genes, and to express this DNA vaccine in HepG2 cells. Methods:The HBV multi-epitope antigen gene was designed and synthesized before it was fused with enhanced green fluorescent protein(EGFP) gene, and cloned into the multi-clone site(MCS) of the eukaryotic expression vector pVAX1. The expressinig units of hIL-12 and siRNA were cloned into the BspH I and Mlu I site of pVAX1 respectively. Then the recombinant plasmid pVAX1-siHBV-HB-EGFP-hIL12 was transiently transfected HepG2 cells. The expression of HBV compound multi-epitope gene was observed through EGFP report gene. The expression of hIL-12 was analyzed by ELISA and the effects of anti-HBV siRNA was confirmed with rtPCR . Results: The analysis of enzyme digestion and sequencing both demonstrated that the trible-expressing HBV DNA vaccine has been constructed successfully. The green fluorescent image was detected in the transfected cells which could confirm the expression of the multi-epitope antigen gene. The amount of hIL-12 secretion was 1289pg/ml in supernatant at 48h after transfection and 1712pg/ml at 72h after transfection. The mRNA amount of HBV S gene, which was the siRNA target, had been obviously knockdown. Conclusion: The DNA vaccine co-expressing the HBV compound multi-epitope antigen gene, the hIL-12 and the siRNA genes was constructed and transiently expressed in HepG2 cells, and siRNA had shown us a good anti-HBV effect. It laid a foundation of further study on anti-HBV effect of the new DNA vaccine.