Objective To clone and express the E2 gene in envelope region of hepatitis C virus and detect the anti E2 in sera of patients with HCV infection,using the purified E2 Protein.Methods The gene fragment of E2 region was obtained from the sera of patients with HCV infection by random primer reverse transcription and polymerase chain reaction(PCR),and then cloned into procaryotic expression system to express and Purify the protein and detect its antigenicity.Results A segment of E2 region gene with correct code reading frame was obtained,its full length was 984 bp and molecular weight of expressed Protein was 38kD.The detection of the protein performed by ELISA with sera from the patients with HCV infection,testified that it had appropriate antigenicity.Conclusion This experiment can provide the fundamental data for researching on the basic characteristics of HCV envelope protein,nuderstanding the significance of serological diagnosis and designing the anti HCV vassine.

The medicinal fungi, which are of great importance in traditional medicine, are facing the problems of wild resources scarcity and low concentration of bioactive compounds. Velvet family and LaeA global regulator play a vital role in secondary metabolism and developmental programs, which are found in a wide variety of fungi ranging from Chytridiomycota to Basidiomycota. This review elaborates the structures and functions between Velvet family and LaeA protein. The Velvet family which shares the Velvet protein domain, including VeA (Velvet), VelB (Velvet like B), VosA (viability of spores A) and VelC (Velvet like C), acts on the regulation function is secondary metabolism and developmental programs such as asexual and sexual development. Furthermore, the function is affected by environmental factors such as light and temperature. LaeA protein which owns S-adenosylmethionine-dependent methyltransferase domain, coordinately regulates development and secondary metabolism by regulating and modifying the Velvet proteins. The regulation of LaeA is mediated by light receptor proteins. Therefore, clarifying the mechanism of Velvet and LaeA proteins in medicinal fungi will pave the way for nurturing medicinal fungi and improving production of bioactive compounds.
Fungal Proteins ; metabolism ; Fungi ; chemistry ; Gene Expression Regulation, Fungal ; Genes, Regulator ; Protein Structure, Tertiary ; Secondary Metabolism ; Structure-Activity Relationship

OBJECTIVETo evaluate the clinical effects of dynamic neutralization system (K-Rod) in treating multisegmental lumbar degenerative disease.

METHODSFrom October 2011 to October 2013, 20 patients with multisegmental lumbar degenerative disease were treated with dynamic neutralization system (K-Rod). There were 8 males and 12 females with an average age of 45.4 years old (ranged from 31 to 65) and an average course of 3.8 years (ranged from 9 months to 6.25 years). All patients had the history of low back and legs pain. Among them, 10 cases were far lateral lumbar disc herniation, 7 cases were lumbar spinal stenosis, 3 cases were lumbar spondylolisthesis (degree I in 2 cases and degree II in 1 case). Every patient had only one responsible segment which causing the symptom would have to be rigidly fixed during operations, and the adjacent intervertebral disc of the responsible segments at least 1 segment has already obvious degenerated. All patients underwent the operation to relieve compressed nerves and reconstruct spinal stability with K-Rod system (the responsible segments were fixed with interbody fusion, and the adjacent segments were fixed with dynamic stabilization). Visual analogue scale (VAS), Japanese Orthopaedic Association Scores (JOA) and Oswestry Disability Index (ODI) were used to evaluate the clinical effects. Imaging data were used to analyze the range of motion (ROM), intervertebral disc height and intervertebral disc signal (according to modified Pfirrmann grading system) in degenerative adjacent segment.

RESULTSAll patients were followed up for more than 1 year, and preoperative symptoms obviously relieved. There were significant differences in VAS, JOA, ODI between preoperative and postoperative (postoperative at 1 week and 1 year) (P<0.05). Radiological examination showed that all responsible segments had already fused, and no looseness, displacement and breakage of internal fixations were found. Postoperative at 1 year, the ROM of adjacent segments were decreased (P<0.05). There was no significant difference in intervertebral disc height between preoperative and postoperative at 1 year (P>0.05). According to modified Pfirrmann grading system to classification for the 25 disks of adjacent segment, 8 disks (32%) got improvement, 15 disks (60%) got no change and 2 disks (8%) got aggravation at 1 year after operation.

CONCLUSIONDynamic neutralization system (K-Rod) combined with interbody fusion could obtain short-term clinical effects in the treatment of multisegmental lumbar degenerative disease.

Adult ; Aged ; Female ; Humans ; Intervertebral Disc Displacement ; surgery ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Range of Motion, Articular ; Spinal Diseases ; surgery ; Spinal Fusion ; methods ; Spinal Stenosis ; surgery ; Spondylolisthesis ; surgery

Adult ; Basic Helix-Loop-Helix Transcription Factors ; metabolism ; Cerebral Ventricle Neoplasms ; metabolism ; pathology ; surgery ; Female ; Follow-Up Studies ; Fourth Ventricle ; Ganglioglioma ; metabolism ; pathology ; surgery ; Glial Fibrillary Acidic Protein ; metabolism ; Humans ; Magnetic Resonance Imaging ; Nerve Tissue Proteins ; metabolism ; Oligodendrocyte Transcription Factor 2 ; Rosette Formation ; Synaptophysin ; metabolism ; Young Adult

OBJECTIVETo preliminarily research the formular about the Traditional Chinese Medicine (TCM) syndrome of adolescent neck pain.

METHODSAn observation table of adolescent neck pain syndromes was formulated,and 1 397 patients with adolescent neck pain were investigated to establish a database of adolescent neck pain. The Descriptive Statistical Analysis and Hierarchical Cluster Analysis were performed by statistical software.

RESULTSTotally 60 TCM symptoms was clustered into 4 TCM syndromes by Hierarchical Cluster Analysis. The expert panel of TCM syndromes preliminarily formulate 4 TCM syndromes of adolescent neck pain by analyzing the result of Cluster Analysis and discussing their clinical experience.

CONCLUSIONAdolescent neck pain is a category of Tendon Trauma's Bi-syndrome of TCM. Ying, Wei, Qi and blood block caused by exopathy, strains, and internal injury is considered as the main pathogenesis of adolescent neck pain. Base on statistical result and expert's opinions, 4 TCM syndromes about adolescent neck pain were formulated: cold-dampness syndrome, dampness-heat blockage syndrome, liver-stagnation and spleen-deficiency syndrome, Qi and Yin deficiency of both heart and kidney syndrome.

Adolescent ; Adult ; China ; epidemiology ; Diagnosis, Differential ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Neck Pain ; diagnosis ; epidemiology ; Young Adult

Objective:To report a case of combined oxidative phosphorylation deficiency 28 (COXPD28) in China, identified the pathogenic mutation and explored the pathogenic mechanism preliminarily.Methods:The clinical characteristics of a patient with COXPD28 were retrospectively analyzed and the pathogenic mutations were identified by mitochondrial gene sequencing and whole exome sequencing. The wild-type and mutant plasmids of pathogenic genes were constructed, and effect of mutation on protein expression by quantitative real-time PCR (qPCR) and Western blot were evaluated. Statistical methods mainly used one-way ANOVA and LSD test.Results:A 21 year old female patient presented with lactic acid poisoning due to repeated chest distress and wheezing since childhood. The sequencing of the whole exon group gene found that solute carrier family 25 member 26 (SLC25A26) gene had a compound heterozygous mutation (c.34G>C, p.A12P; c.197C>A, p.A66E), which was the first report in China. In vitro function test showed that the expression levels of SLC25A26 mRNA and S-adenosylmethionine carrier (SAMC) protein in cells transfected with SLC25A26 mutant plasmid were significantly lower than those transfected with wild type plasmid. The p.A66E mutant plasmid reduced the expression level of SLC25A26 mRNA and SAMC protein to 6% and 26% of wild type plasmids respectively (both P<0.001), while p.A12P mutant plasmid decreased to 62% and 82% of wild type plasmids respectively ( P<0.001, P=0.044). When the double mutant (p.A66E+p.A12P) plasmids were co-transfected, the expression levels of SLC25A26 mRNA and SAMC protein decreased to 47% and 57% of the wild type plasmids, respectively ( P<0.001, P=0.001). Conclusion:The pathogenic mutation gene of this patient with COXPD28 is SLC25A26 gene mutation (p.A66E, p.A12P), which causes the decrease of SLC25A26 expression level, mitochondrial oxidative phosphorylation dysfunction, and induces COXPD28.

Transcription factor is one of the key factors in the regulation of gene expression at the transcriptional level. It plays an important role in plant growth, active components biosynthesis and response to environmental change. This paper summarized the structure and classification of bHLH transcription factors and elaborated the research progress of bHLH transcription factors which regulate the active components in plants, such as flavonoids, alkaloids, and terpenoids. In addition, the possibility of increasing the concentration of active substances by bHLH in medicinal plants was assessed. The paper emphasized great significance of model plants and multidisciplinary research fields including modern genomics, transcriptomics, metabolomics and bioinformatics, providing the contribution to improve the discovery and function characterization of bHLH transcription factors. Accelerating the research in the mechanism of bHLH transcription factors on the regulation of active components biosynthesis will promote the development of breeding and variety improvement of Chinese medicinal materials, also ease the pressure of resources exhaustion of traditional Chinese medicine home and abroad.
Alkaloids ; biosynthesis ; Basic Helix-Loop-Helix Transcription Factors ; chemistry ; classification ; genetics ; metabolism ; Flavonoids ; biosynthesis ; Plants, Medicinal ; genetics ; metabolism ; Terpenes ; metabolism

AIM To explore the effects of Zishui Qinggan Decoction on the mouse model of depression induced by chronic restraint stress(CRS)via ERK/GSK3β/CREB/BDNF signaling pathway.METHODS Except for those of the blank group,the mice of other groups were induced into depression models by CRS,and divided into the model group,the fluoxetine hydrochloride group(10 mg/kg)and the low,medium and high dose Zishui Qinggan Decoction groups(8.835,17.670 and 35.340 g/kg)for the corresponding drug intervention and simultanous CRS treatment.The mice had their sugar water preference experiment and behavior experiment on the 7th and 14th day after administration;the observation of the hippocampal morphological changes by HE staining,the detection of the superoxide dismutase(SOD)activity and malondialdehyde(MDA)level in serum by kits,the detection of levels of serum 5-hydroxytryptamine(5-HT),tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)by ELISA,the detection of the hippocampal mRNA expressions of BDNF,TNF-α and IL-1β by RT-qPCR method,and the detection of the hippocampal protein expressions of ERK1/2,p-ERK1/2,GSK3β,p-GSK3β,CREB and BDNF by Western blot method 14 days after administration.RESULTS Compared with the model group,after 14 days of administration,both fluoxetine hydrochloride group and medium-dose Zishui Qinggan Decoction group displayed increased preference rate of sugar water(P<0.01),shortened immobility time of tail suspension and forced swimming(P<0.01),improved hippocampal damage of nerve cells,decreased levels of serum MDA,TNF-α and IL-1β(P<0.05,P<0.01),increased SOD activity and 5-HT level(P<0.05,P<0.01),decreased hippocampal mRNA expressions of TNF-α and IL-1β(P<0.01),and decreased expressions of BDNF mRNA and p-ERK1/2,p-GSK3β,CREB and BDNF proteins(P<0.05,P<0.01).CONCLUSION Zishui Qinggan Decoction can improve the depression-like behaviors in mice exposed to CRS,and its mechanism may be related to the regulation of hippocampal ERK/GSK3β/CREB/BDNF signaling pathway.

ObjectiveTo describe the modified (narrowed) gastric tube and two methods on the establishmentof enteral feeding pathway in esophagectomy.MethodsFrom July 2003 to April 2006,we made traditional gastric tube for patients underwent esophagectomy to substitute the dissected esophagus while from May 2006 to November 2009,we made modified gastric tube for the same purpose.From July 2003 to November 2009,enteral nutrition tubes for all these patients are placed intraoperatively by two different ways,which made early post-operative nutrition support possible for these patients.ResultsNo perioperative death occurred among the patients.The length of the modified gastric tube ensured the anastomosis by circular stapler at the apex thorax or in the bases of cervical region.No statistical differences were found between the two group in terms of survival rate.The intraoperative establishment of enteral nutrition pathway ensured the early enteral nutrition support after the operation.ConclusionThe utility of the modified gastric tube extends the length of gastric tube to make mechanical anastomose easier and safer; meanwhile,the intraoperative establishment of enteral nutrition pathway ensures the early enteral nutrition support after the operation.

A simple and quick method is described for the determination of ferulic acid, senkyunolide I, senkyunolide H, senkyunolide A and ligustilide in rhizomes of Ligusticum chuanxiong. The 5 active ingredients in the sample was extracted using 40% ethanol and analyzed by reversed-phase high performance liquid chromatography (HPLC). Chromatography separation was performed using Agilent 1100 series HPLC system with a Symmetry C18 column and gradient elution with a mixture of three solvents : solvent A, acetonitrile, solvent B, methanol and solvent C, 1% aqueous acetic acid, 0 min to 5 min A: B: C 20: 40: 40, 5 min to 30 min A: B: C 60 to 100 : 0 : 40 to 0. The effluent was monitored using a VWD detector set at 321 nm (0-4.3 min) and 275 nm (4.31-30 min). The flow rate was set at 1 mL x min(-1) and the injection volume was 10 microL. The column temperature was maintained at 35 degrees C. The calibration curve was linear (r > or = 0.99) over the tested ranges. The average recovery was 94.44%-103.1% (n = 6). The method has been successfully applied to the analysis in different harvest periods of L. chuanxiong samples. In this paper, single-factor randomized block design to study the 5 components content of L. chuanxiong on ten collecting stages. For the L. chuanxiong collected from April 15th to May 30rd, the content of 5 ingredients increased primarily, and then decreased. Determine the appropriate harvest time has important significance to the promotion of the quality of L. chuanxiong.
4-Butyrolactone ; analogs & derivatives ; analysis ; Acetic Acid ; chemistry ; Acetonitriles ; chemistry ; Benzofurans ; analysis ; Chromatography, High Pressure Liquid ; methods ; Coumaric Acids ; analysis ; Ligusticum ; chemistry ; Methanol ; chemistry ; Solvents ; chemistry ; Time Factors