The alpine plant Gentiana robusta is an endemic species to the Sino-Himalayan subregion. Also, it is one of the original plants used as traditional Tibetan medicine Jie-Ji. We sequence the nuclear ribosomal internal transcribed spacer (ITS) regions, matK, rbcL, rpoC1, trnL (UAA), psbA-trnH, atpB-rbcL, trnS( GCU)-trnG(UCC), rpl20-rps12, trnL(UAA)-trnF( GAA) fragments of cp DNA in both G. robusta and such relative species as G. straminea, G. crassicaulis and G. waltonii. With Halenia elliptica as the outgroup, molecular systematic analysis reveals that G. robusta is a natural hybrid. G. straminea is the mother of hybrids, but the father is not very clear. In addition, the molecular markers for distinguishing G. robusta from the parental species or closely related species are identified, respectively. Our studies may provide valuable reference for the species identifications of medicinal plants with complex genetic backgrounds.
DNA Barcoding, Taxonomic ; DNA, Plant ; genetics ; Gentiana ; classification ; genetics ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; genetics ; Plants, Medicinal ; classification ; genetics

AIM: To investigate the feasibility and clinical effect of punctoplasty by using trabeculectomy punch combined with a novel RS tube for the treatment of punctal stenosis.METHODS: Totally 39 patients (39 eyes) with punctual stenosis were selected from October 2013 to October 2015 in the Second People`s Hospital of Foshan.All patients underwent punctoplasty by using trabeculectomy punch combined with a novel RS tube.These tubes were removed at 3mo after operation.A follow-up of 6mo was taken for final analysis.The fluorescein dye disappearance test score was recorded before the operation and at 1,3 and 6mo after the extubation.The curative effect of the operation at 6mo after the extubation was assess.RESULTS: Fluorescein dye disappearance test: the scores at 1,3 and 6mo after the extubation all decreased compared with the preoperative ones.The difference was statistically significant(P<0.05).At the last following up, 35 eyes (90%) were cured completely, 4 eyes (10%) were improved significantly, no patients recurred.Effective rate was 100%.No serious intraoperative and postoperative complications happened.CONCLUSION: Punctoplasty by using trabeculectomy punch combined with novel RS tubes is a safe and effective method for the punctul stenosis, which is easy to perform, with high success rate.

Background The operation mode of automobile manufacturing industry (AMI) makes workers have different degrees of occupational stress and burnout, which may lead to negative emotions and depressive symptoms. Objective To study the relationship between occupational stress, job burnout, and depressive symptoms in AMI workers. Methods In this study, 1300 workers from a Guangzhou AMI company were selected as subjects by cluster random sampling method. Occupational stress, job burnout, and depressive symptoms of the workers were assessed by using the Effort-Reward Imbalance (ERI) questionnaire, the Maslach Burnout Inventory general survey questionnaire, and the Patient Health Questionnaire-9, respectively. Hierarchical regression was used to analyze the effects of occupational stress and job burnout on depressive symptoms in AMI workers. Mediating effect model was used to analyze the mediating effect of job burnout on the relationship between occupational stress and depressive symptoms. Results There were 1300 questionnaires distributed, 1228 valid questionnaires collected, with a 94.5% recovery rate. The ERI ratio of 1228 AMI workers was 1.06±0.72, and the positive rate of occupational stress was 37.3% (458/1228). The score of job burnout was 2.18±1.37, and the positive rate of job burnout was 62.6% (769/1228). The score of depressive symptoms was 10.27±6.42, and the positive rate of depressive symptoms was 47.1% (578/1228). The dimensional scores of effort and over-commitment in occupational stress as well as emotional exhaustion and depersonalization in job burnout of AMI workers were positively correlated with the depressive symptom scores (r_s=0.415, 0.571, 0.573, 0.593, P＜0.05). The dimensional scores of reward and personal achievement were negatively correlated (r_s=−0.454, −0.339, P＜0.05). The percentages of variance in depressive symptoms score explained by occupational stress and job burnout were 26.7% and 16.6%, respectively. Job burnout had a partial mediating effect between the three dimensions of occupational stress and depressive symptoms, and the mediating effect values were −0.2832 (95%CI: −0.3250– −0.2434), 0.3553 (95%CI: 0.3071–0.4041), and 0.4193 (95%CI: 0.3681–0.4725), respectively. Conclusion AMI workers' occupational stress affects job burnout, but also indirectly affects depressive symptoms. Job burnout partially mediates the association between occupational stress and depressive symptoms. Reducing occupational stress and burnout levels of AMI workers may alleviate depressive symptoms.

The skin ulceration syndrome of sea cucumber is a kind of desease induced by bacterium.In order to investigate the bacterium of infected sea cucumber and detect the N-acyl-homoserine lactones(AHLs) se-cretion of the bacterium,7 bacterial strains were isolated from the infected sea cucumber.These strains were identified by physiological-biochemical characteristics and 16S rDNA sequence.Results show that strain C6 belongs to Tenacibaculum,strain 4 belongs to Shewanella putrefaciens group,strain TB belongs to Vibrio,strain BP2,BP3,BP4 and BP6 belong to Pseudoalteromonas,respectively.AHLs were detected with strain Agrobacterium tumefaciens KYC55.Among these bacterial strains,strain C6,4,TB,BP3 and BP4 can se-cret AHLs,while strain BP2 and BP6 can’t.And the AHLs activity differs,from the highest to the lowest are 4,TB,BP4,BP3 and C6.

OBJECTIVETo investigate the effects of single heat stress treatment on spermatogenic cells in mice.

METHODSWe randomly divided 36 C57 male mice into a control and a heat stress treatment group and submerged the lower part of the torso in water at 25 °C and 43 °C, respectively, both for 15 minutes. At 1, 7, and 14 days after treatment, we obtained the testicular organ indexes, observed the changes in testicular morphology by HE staining, and determined the location and expression levels of the promyelocytic leukemia zinc finger (PLZF) and synaptonemal comlex protein-3 (SCP-3) in the testis tissue by immunohistochemistry and Western blot.

RESULTSThe testicular organ index was significantly lower in the heat stress treatment than in the control group (P < 0.05). Compared with the controls, the heat shock-treated mice showed loosely arranged spermatogenic cells scattered in the seminiferous tubules at 1 day after heat stress treatment, atrophied, loosely arranged and obviously reduced number of spermatogenic cells at 7 days, and relatively closely arranged seminiferous tubules and increased number and layers of spermatogenic cells at 14 days. The number of SCP-3 labelled spermatocytes obviously decreased in the heat stress-treated animals at 1 and 7 days and began to increase at 14 days. The PLZF protein expression was significantly reduced in the heat stress treatment group at 1 day as compared with that in the control (0.19 ± 0.12 vs 0.64 ± 0.03, P < 0.01), but elevated to 0.77 ± 0.02 at 7 and 14 days, even remarkably higher than in the control animals (P < 0.01).

CONCLUSIONHeat stress treatment can induce short-term dyszoospermia in mice, which can be recovered with the prolonged time after treatment.

Animals ; Blotting, Western ; Hot Temperature ; Immunohistochemistry ; Male ; Mice ; Nuclear Proteins ; metabolism ; Promyelocytic Leukemia Protein ; Seminiferous Tubules ; cytology ; Spermatocytes ; cytology ; pathology ; Testis ; metabolism ; Transcription Factors ; metabolism ; Tumor Suppressor Proteins ; metabolism

OBJECTIVETo express and purify the recombinant human bone morphogenetic protein-2 mature peptide (rhBMP-2m) in prokaryotic system and to develop highly-specific monoclonal antibodies.

METHODSAn engineered E. coli strain expressing rhBMP-2m was fermented. The bacterial cells were firstly lysed and then the rhBMP-2m inclusion bodies were isolated by centrifugation. After the inclusion bodies had been solubilized by high-concentration denaturing agents, denatured rhBMP-2m was purified by cation ion-exchange chromatography. Biologically active rhBMP-2m was obtained by refolding of purified denatured rhBMP-2m through direct dilution. The refolded rhBMP-2m was used to immunize Balb/c mice to develop anti-rhBMP-2m monoclonal antibodies using classic hybridoma technique.

RESULTSrhBMP-2m with a purity greater than 95% was obtained on reduced SDS-PAGE. The refolded rhBMP-2m was measured to be bioactive by the induction of alkaline phosphatase activity in MC3T3-E1 cells. Two hybridoma cell lines that stably secreted anti-rhBMP-2m antibody were developed from the immunized mice.

CONCLUSIONBioactive rhBMP-2m protein and its monoclonal antibodies were successfully prepared, which will provides a solid base for future studies on rhBMP-2.

Animals ; Antibodies, Monoclonal ; biosynthesis ; Bone Morphogenetic Proteins ; biosynthesis ; immunology ; Escherichia coli ; metabolism ; Humans ; Mice ; Mice, Inbred BALB C

Background Placement of an orbital implant is a main way to prevent orbital atrophy with aging.But its mechanism is under clear.Researchs showed that bone growth factors play important role during the development and repair of bone,especially transforming growth factor-β1(TGF-β1).Objective Present study was to investigate the expression of TGF-β1 protein in orbital bone after enucleation or enucleation with placement of an orbital implant and its function in the mechanisms of preventing and treating the orbital malformed development after enucleation with placement of an orbital implant.Methods Twenty-one age- and weight-matched New Zealand white young rabbits were randomly divided into the enucleation,implant and control groups,and each group including seven rabbits.Eyeball nucleation surgery was performed in the left eyes of 7 1-month-old rabbits,and a spherical orbital implant was inserted after enucleation of the left in matched rabbits in implant group.The left eye of normal rabbits served as controls.The rabbits were sacrificed in 1 month after surgery.The expression of TGF-β1 protein in the left orbital bone was detected using enzyme immunoassay and FITC labelling immunoassay technique in the sections of zygomatic bones.The content of TGF-β1 protein in the left orbital bone tissue was measured by ELISA method.This use of animals complied with the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results The height and width of orbital in enucleation group were significantly lower than those of implant and normal control groups(height:P=0.00,P=0.00;width:P =0.00,P=0.00).The positive bone cells of both enzyme immunoassay and FITC staining were increased in the implant and control groups in comparison with enucleation group,but the positive response intensity for TGF-β1 was resembled between implant group and control group.ELISA result revealed that the content of TGF-β1 protein in bone tissue was significantly lower in the enucleation group than in implant and control groups(P=0.00,P=0.00).The expression and content of TGF-β1 protein in bone tissue is similar between the implant group and the control group(P=0.41). Conclusion The experiment results indicate that TGF-β1 protein participate in the orbital development.TGF-β1 played important role in the prevention and treatment of enucleation-induced orbital malformation in the eye with placement of an orbital implant.

0.05).Conclusion MK mRNA overexpresses in the breast cancer tissues.It might be considered to be a reference indicator for deter- mining the angiogenesis and invasion of breast carcinoma.

Aim To investigate the influence of Shen-mai injection ( SMI ) on the expression of cytochrome P450(CYP450) system in rat′s hearts. Methods Rat hearts were prepared after a fourteen-day continuous administration of SMI. The expression of several CYP genes, ANP, BNP and EPHX2 were measured by qPCR. Results SMI induced the increase in the ex-pression of other CYP genes except CYP2 B1、CYP4 A3 and CYP4 F6;HSI caused an induction of CYP2 E1 , CYP4A3,CYP4F1 and EHPX2 as compared with the control. In addition, there was a significant induction of ANP, BNP and EHPX2 and a significant inhibition of CYP2B1 and CYP2C11 after treated with MDI. Conclusion Although there is no significant change in the gene expression of CYP2 B1 after the treatment with SMI, but there is a general trend of induction, and MDI shows a significant inhibition of CYP2 B1 , therefore HSI has greater effect on CYP 2 B 1 than MDI . SMI causes a significant induction of CYP2 E1 , CYP4F1 and EHPX2 , similarly there is an induction of CYP2E1,CYP4F1 and EHPX2 by HSI and MDI, indi-cating that Hongshen and Maidong are both involved in the induction. MDI has a greater inductive effect than HSI on ANP and BNP. SMI is widely used for the treatment of cardiovascular diseases due to its regula-tion of CYP2J3、ANP and BNP mRNA expression.

Objective: To establish a method for isolation and purification of fetal membrane derived adherent cells (FMDACs) , and investigate their biological characteristics. Method: FMDACs were isolated with trypsin inducing and cultured in vitro. FMDACs were induced to differentiate into osteoblasts and adipocytes. FACS and immunocytochemistry technique were used to examine the cell surface antigen. The genetic stability was verified by karyotype analysis. Results: FMDACs were successfully isolated and expanded in vitro. They had strong proliferative ability. FMDACs were positive for CD44 and CD29, but negative for CD34, CD14 and CD45. FMDACs were differentiated into osteoblasts and adipocytes after inducement. The karyotype was stable in the sixth-passaged FMDACs and the tumorigenicity was not found. Conclusion; FMDACs have the possibility of multipotent stem cells, which have strong capacities of self-renewal and multidirectional differentiation. The genetic background of FMDACs is stable. FMDACs may be used as a kind of novel seed cells for tissue engineering.