The bioreactor production of recombinant Lateolabrax japonicus growth hormone (rljGH) expressed intracellularly by Pichia pastoris was investigated. A strategy of feeding methanol at the exponential rate was established and the effect of specific growth rate on the rljGH production was examined. The results indicated that the average specific production rate increased and the rljGH production duration decreased as the specific growth rate increased. The maximum specific rljGH production (0.58 mg/g WCW) was achieved at a specific growth rate of 0.029/h. The effect of supplementing ammonium sulfate, peptone and yeast ex- tract on the rljGH production was further investigated. The results indicated that the effects of ammonium sulfate and peptone were not significant. Supplementing yeast extract of 2.5 g/L was advantageous for the rljGH production. The duration of the rljGH production was increased to 23 h from 17 h and the fermenta- tion stability of run-to-run could be improved.

Objective To study the expression of Rho kinase and its functional activation in lung tissue from hypoxic pulmonary hypertension(HPH) rat model,and the effects of fasudil on HPH.Methods Seventy-two male Spraque-Dawley rats were randomly divided into the control group,hypoxic model group,and fasudil-intervention group[group with hypoxia and fasudil for 15 mg/(kg?d)],respectively.Mean pulmonary arterial pressure(mPAP) and right ventricle hypertrophy index(RVHI) were measured.Expression of Rho kinase mRNA and protein were examined by reverse transcriptase-polymerase chain reaction(RT-PCR) and Western blot,respectively.The phosphorylation of binding subunit of myosin phosphatase(MBS)-a substrate of Rho kinase was detected by Western blot and defined,as the mark of functional activation of the kinase.Results The expression of Rho kinase mRNA in hypoxic model group was markedly upregulated even before the onset of the third day after the experiment(HPH),and it was much lower in rats of fasudil group than that of hypoxic model group.The phosphorylation of MBS was significantly higher in hypoxic model group than that in control group,and it was positively correlated with the mPAP and RVHI(all P

OBJECTIVEThe study was to verify the feasibility of an improved method using reverse flow island flap nourished by the sural nerve nutrition vessel to repair severe frostbite of feet.

METHODSAt the proximal end of the principal flap, an auxiliary triangular skin flap of 6.5 - 7 cm in length was designed in order to cover the pedicle of the principal flap. This operation was performed on 13 patients (21 feet) with frostbite.

RESULTSAll the flaps survived well. Postoperative follow-up for 5 - 18 months demonstrated satisfactory results in all the cases. No ulcer happened.

CONCLUSIONSThe method is helpful to prevent constriction of the pedicle and ensure blood supply of the flap. It is an ideal treatment for severe frostbite of feet.

Adult ; Female ; Foot Injuries ; surgery ; Frostbite ; surgery ; Humans ; Male ; Reconstructive Surgical Procedures ; methods ; Skin Transplantation ; Surgical Flaps ; blood supply ; innervation ; Young Adult

Objective To evaluate the risk of plague occurrence via surveying and analyzing indoor rat density and flea index in natural villages having previous plague experience. Methods During August to September 2007, 30 natural villages experiencing previous plague were selected based on the surveillance data, and then all households were coded with numbers and 20 households in each village were randomly selected via computer. Cages and sticky papers were set in 600 selected households to capture rats and fleas. Rat density, flea prevalence, flea index and median were estimated. Results One hundred thirty-three Rattus flavipectus and 33 Suncus murinus were caught and averaged rat density was 2.8 rats per one hundred cage. nights (166/6000), the median was 5 rats each village. One hundred and one mice infected fleas, flea prevalence on rats was 60.8% (101/166), 296 Xenopsylla cheopis and 48 Leptopsylla segnis were collected. Rat flea index was 2.1 fleas per rat (344/166). A total of 315 dissociated flea was caught, average dissociated flea index was 0.026 fleas per sticky paper (315/11888). The median was 5.5 dissociated fleas per village. Of dissociated fleas, Ctenocephalides felis felis (205) and Xenopsylla cheopis (103) accounted for 97.8% (308/315). The proportion for species of the rat flea and the dissociated flea was different(Fisher test: P < 0.01). The rat flea was significantly associated with the rat density(r = 0.68, P < 0.01), but the dissociated flea was significantly associated with neither the rat density(r = -yield than fried wheat batter(χ2 = 5.59, P < 0.05). Conclusions In these villages having previous plague experience of Lianghe County, Rattusflavipectus was dominant species of indoor rats, Xenopsylla cheopis and Ctenocephalides felis felis were dominant species of rat flea and dissociated flea, respectively. Mengsong, Bangdu, and Tangjiatun village had potential risk of plague emergence.

A new modification method for glass slides was developed and applied to make ThinPrep Pap smears,in order to increase the adhesion ability of cervical exfoliative cells.3-glycidyloxypropyl trimethoxysilane (GOPS) was coated on the glass slides firstly on the slides,then poly-L-lysine (PLL)was covalently modified onto the above epoxy-terminated slides to form GOPS-PLL double decorated slides.The modified slides were characterized using X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM).The cell adhesion ability effect was tested and compared with traditional PLL coated slides by fixing the cervical exfoliative cells on the double adorned slides.The control test was conducted by the bare glass slides unmodified.The cell morphology of cervical exfoliative cells adhered on different slides was observed under the microscope after Papanicolaou staining.The number of cervical exfoliative cells on the unmodified slides,PLL coated slides and GOPS-PLL coated slides was 1030±300,3283±226 and 4119±280 (n=12),respectively.The data among the three different modification methods showed significant differences (one-way analysis of variance,ANOVA test,P＜ 0.05).The cell capturing effect of the GOPS-PLL slide was the best among the three different modified slides.In addition,the GOPS-PLL slide could enhance the uniformity of the adhered cells and be widely applied to the ThinPrep system for cervical carcinoma screening to increase the accuracy rate of diagnosis.

Xuefu Zhuyu decoction (XFZYD) is a famous traditional Chinese medicine (TCM) formula, is widely used in the treatment of cardiovascular and cerebrovascular diseases in China over one hundred years. But its effect on antioxidant and drug-metabolizing enzymes are unknown. This study was to observe the effects of Xuefu Zhuyu decoction (XFZYD) on the activities of antioxidant and drug metabolism enzymes (DMEs) in liver of rats. Male SD rats, treated with XFZYD at the dosage of 3.51, 7.02 and 14.04 g x kg(-1) per day for 15 days, serum were collected, tissue fluid, cytosols and microsomes isolated from liver tissues were prepared by centrifugation according to the standard procedure, the activities of antioxidant enzymes and drug-Metabolizing Enzymes were determined by UV-V is spectrophotometer. In serum, the activities of AST was not significantly affected by the treatment with XFZYD, at the high- est dose, the levels of ALT, Cr and BUN were significantly decreased (P < 0.05). GPX were significantly increased at the dose of 7.02, 14.04 g x kg(-1) (P < 0.05), CAT were significantly increased at the highest dose (P < 0.05). T-SOD was not significantly af- fected by this treatment. In the liver tissue, GPX was significantly increased at the dose of 3.51, 7.02 g x kg(-1) (P < 0.05), GST, CAT and T-SOD were not significantly affected following this treatment. In cytosols, GST was significantly increased at the dose of 3.51 g x kg(-1) (P < 0.05), T-SOD was remarkable induced at the dose of 3.51 and 7.02 g x kg(-1) (P < 0.05). In microsomes, XFZYD had no significant effect on Cytochromeb5, NADPH-Cytochrome P450 reductase, CYP3A, CYP2E1 and UGT, XFZYD significantly in- duced GST at the dose of 3.51 and 7.02 g x kg(-1) (P < 0.05), and the level of GSH were significantly increased by XFZYD at the dose of 3.51, 7.02 and 14.04 g kg(-1) (P < 0.05). These findings suggest XFZYD can induce the activities of GPX, CAT, SOD, GST and increase GSH level in liver of rats, which indicate XFZYD may have detoxification and antioxidant functions.
Animals ; Antioxidants ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Inactivation, Metabolic ; drug effects ; Liver ; drug effects ; enzymology ; Male ; Rats ; Rats, Sprague-Dawley

Mitogen-activated protein kinases (MAPKs) are important signaling transduction components well conserved in eukaryotes and play essential roles in various physiological, developmental and hormonal responses in plant. In the present study, a MAPK gene, designated as DoMPK4 (GenBank accession No. JX297597), is identified from a rare endangered medicinal orchid species D. officinale using the reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) methods. The full length cDNA of DoMPK4 is 1 518 bp in length and encoded a 369 aa protein with a molecular weight of 42.42 kD and an isoelectric point of 5.55. DoMPK4 protein contained a serine/threonine protein kinase active site (158-170), a MAP kinase site (71-174), and eight conserved motifs. DoMPK4 had a transmembrane (214-232) but no signal peptide. Multiple sequence alignment showed that DoMPK4 shared high identities (74.9%-80.6%) with MAPK proteins from various plants. Phylogenetic analysis demonstrated that DoMPK4 belonged to group A of the MAPK evolutionary tree, and is closely related to monocots. Real time quantitative PCR (qPCR) analysis revealed that DoMPK4 is differentially expressed among the five organs including leaf, stem, root, seed, and protocorm-like body (PLB). The transcription level of DoMPK4 is the highest in the PLBs with 17.65 fold, followed by seeds, roots, and stems with 5.84, 2.28, and 1.64 fold, respectively. The progressive enhancement of DoMPK4 transcripts in the developing PLBs compared to that in the germinating seeds, suggests a role of DoMPK4 during the development of embryogenic PLBs formation in D. officinale.
Amino Acid Sequence ; DNA, Complementary ; genetics ; DNA, Plant ; genetics ; Dendrobium ; enzymology ; genetics ; Gene Expression Regulation, Plant ; Mitogen-Activated Protein Kinases ; genetics ; metabolism ; Phylogeny ; Plant Leaves ; metabolism ; Plant Proteins ; genetics ; metabolism ; Plant Roots ; metabolism ; Plant Stems ; metabolism ; Seeds ; metabolism ; Sequence Alignment

Objective To determine the effect of fibroblast growth factor (FGF) on endothelia cell (EC) proliferation in vitro,TNP-470 and dexamethasone (Dex) and explore the possible mechanisms of FGF stimulating EC proliferation. Methods Cultured human umbilical vein endothelial cell (HUVEC) was divided into two groups: control group (Group DMEM) and experimental groups (Group FGF,Group Dex,Group TNP-470,Group FGF+Dex,Group FGF+TNP-470). The EC proliferation was quantified by a colorimetric assay using MTT reagent,the relative cell numbers at every time point of EC cycle analyzed with flow cytometer,and the expression of nuclear factor-kappa B p65 (NF-?B p65) and ki-67 detected with immunohistochemical method of SABC. Results FGF stimulated significantly EC proliferation,raised proliferation index (PI) and enhanced the expressions of NF-?B p65 and ki-67 in nuclear. TNP-470 reduced PI and TNP-470 and Dex suppressed EC proliferation and finally decreased the expressions of NF-?B p65 and ki-67 in nuclear. FGF could facilitate the inhibitory effect of TNP-470 and Dex. Conclusions The possible mechanism of EC proliferation stimulated by FGF is that FGF can activate NF-?B to promote the synthesis of DNA and EC mitosis. TNP-470 and Dex suppress proliferation and differentiation of EC but FGF may reverse such suppressive effect.