Chitinases genes from Metarhizium anisopliae which is an important entomopathogenic fungus were considered one of the key factors to invade their hosts. One Metarhizium anisopliae HN1 strain was isolated and screened. A chitinase gene was amplified by RT-PCR from Metarhizium anisopliae HN1, The whole length of this gene was 1275bp,and the nucleotide sequence of the gene was 96% similarity to that of the M. anisopliae E6 accessed in GenBank ( AF02749). The gene has been registered in GenBank and its accession number is DQ011865. The gene was subcloned into prokaryon expression vector pET-22b( + ), transformed this recombinant expression plasmid into E. coli strain BL 21 and effective expressed. The SDS-PAGE analysis indicated that the recombinant protein was 42kDa which is same to the reported article. The expression level of recombinant protein was about 63. 3% of whole expressed proteins , And when recombinant E. coli were crushed by freeze and supersonic wave , the activity assay indicates that the chitinase expressed in bacteria possesses biological activity.

OBJECTIVE:To investigate the relationship between Tacrolimus blood concentration and effectiveness after renal transplantation and to find factors affecting Tacrolimus blood concentration METHODS:Steady-state trough concentrations of Tacrolimus were collected from 80 renal transplantation patients RESULTS:The effective blood concentration of Tacrolimus was associated with course of treatment CONCLUSIONS:The therapeutic concentration of Tacrolimus was between 12ng/ml and 15ng/ml during the first 2 weeks after renal transplantation,then should be 8ng/ml～15ng/ml

In the present study, terminal-restriction fragment length polymorphism (T-RFLP) technique was applied to assess the diversity and tissue distribution of the fungal endophyte communities of Alpinia officinarum collected from Longtang town in Xuwen county, Guangdong province, China, at which the pharmacological effect of the medicine plant is traditional considered to be the significantly higher than that in any other growth areas in China. A total of 28 distinct Terminal-Restriction Fragment (T-RFs) were detected with HhaI Mono-digestion targeted amplified fungal nuclear ribosomal internal transcribed spacer region sequences (rDNA ITS) from the root, rhizome, stem, and leaf internal tissues of A. officinarum plant, indicating that at least 28 distinct fungal species were able to colonize the internal tissue of the host plant. The rDNA ITS-T-RFLP profiles obtained from different tissues of the host plant were obvious distinct. And the numbers of total T-RFs, and the dominant T-RFs detected from various tissues were significantly different. Based on the obtained T-RFLP profiles, Shannon's diversity index and the Shannon's evenness index were calculated, which were significantly different among tissues (P < 0.05). Furthermore, two types of active chemicals, total volatile oils by water vapor distillation method and galangin by methanol extraction-HPLC method, were examined in the each tissue of the tested plant. Both of tested components were detected in all of the four tissues of the medicine plant with varying contents. And the highest was in rhizome tissue. Correlation analysis revealed there were significant negative correlations between both of the tested active components contents and calculated Shannon's diversity index, as well as the Shannon's evenness index of the fungal endophyte communities of the host plant (P = 0, Pearson correlation coefficient ≤ -0.962), and significant positive correlations between both of the tested active components contents and 325 bp dominant T-RF linkage to Pestalotiopsis (P = 0, Pearson correlation coefficient ≥ 0.975). In conclusion, A. officinarum is colonized by diverse fungal endophytes communities. The diversity of the fungal endophytes was found in the A. officinarum varied with differences of the tissue types of the host plants and was closely correlated with the accumulation of main active components, total volatile oils and galangin contents in the host plant tissue.
Alpinia ; chemistry ; microbiology ; Biodiversity ; China ; DNA, Fungal ; genetics ; DNA, Ribosomal ; genetics ; Drugs, Chinese Herbal ; analysis ; Endophytes ; classification ; genetics ; growth & development ; isolation & purification ; Fungi ; classification ; genetics ; growth & development ; isolation & purification ; Phylogeny ; Plants, Medicinal ; chemistry ; microbiology ; Polymorphism, Restriction Fragment Length

Objective To evaluate the impact of postoperative adjuvant chemotherapy on prognosis of patients with advanced rectal carcinoma using a meta-analysis.Methods We searched PubMed to identify literature comparing observation with adjuvant chemotherapy after neoadjuvant chemoradiotherapy and surgery for patients with advanced rectal carcinoma.Data were analysed using Revman 5.0 statistical software.Results Nine trials were included consisting of 6 212 patients:3 421 patients received adjuvant chemotherapy and 2 791 patients did not.The age ranged from 55.6 to 68 years.Adjuvant chemotherapysignificantly affects overall and disease-free survival (P =0.002 and P =0.000 5 respectively) of patientswho had received neoadjuvant chemoradiotherapy.The subgroup analysis which originate from insufficient data reveales ypT0-2 patients and ypT3-4 patients can't benefit from adjuvant chemotherapy.Conclusion Postoperative adjuvant chemotherapy can improve the survival of advanced rectal carcinoma patients after neoadjuvant chemoradiotherapy,but can't increase the survival of pathology complete respone and yPN + patients.

Staphylococcus aureus ( S. aureus ) osteomyelitis, a significant complication for patients un-dergoing fracture fixation, is a great challenge for orthopaedic surgeons due to its extreme difficulty in mangae-ment. Animal models play an important role in exploring the pathogenesis of osteomyelitis and determining the efficacy of prophylactics and therapeutic treatment. To help understand current animal models of S. aureus os-teomyelitis, we conduct a systematic search to identify animal experiments that have investigated the management of S. aureus osteomyelitis. Experimental studies are categorized by animal species and are further classified by the setting of infection. Study methods are summarized and the advantages and disadvantages of each species and model are discussed.

Objective:To investigate the effects of cyclic tensile stress on the function and degeneration of nucleus pulposus cells.Methods:The human primary nucleus pulposus cells were isolated and cultured. The cyclic tensile stress (100 000 μ?, 10% tensile strain, 0.1 Hz, 8 640 cycles) was loaded on the cells for 24 h. The proliferation of the cells was examined by MTT method. The cell cycle and apoptosis were detected through flow cytometry. Gene expression profile chip was used to detect the differentially expressed genes between the tensile stress group and control group. The function of these gene was analyzed by bioinformatics. The expression of inflammatory related factors, TGF-β, matrix degrading enzymes and extracellular matrix molecules were examined by qRT-PCR.Results:The cyclic tensile stress significantly promoted proliferation and cell cycle of nucleus pulposus cells. The cell percentage of S phase ( t=5.336, P<0.05) and G2/M phase ( t=7.288, P<0.01) was significantly different between the tensile stress group and control group. The cyclic tensile stress inhibited apoptosis of nucleus pulposus cells (8.56%±0.48% vs 10.63%±0.32%, t=4.474, P<0.05). A total of 866 differentially expressed genes were detected. Gene ontology analysis showed the roles of these genes in cells including focal adhesion, extractable matrix, membrane raft, condensed chrome kinetochore, cytoskeleton, etc. The cyclic tensile stress significantly affected the mRNA expression of inflammatory related factors, TGF-β genes, matrix proteinase and extracellular matrix molecules. Compared with the control group, the mRNA expression of inflammatory related factors IL15 ( t=5.379, P<0.05), IGF1 ( t=5.454, P<0.05) and IGFBP7 ( t=13.57, P<0.01) were significantly decreased in the tensile stress group; The mRNA expression of TGF-β genes TGFB1 ( t=6.931, P<0.05), TGFB2 ( t= 15.56, P<0.01) and TGFB3 ( t=7.744, P<0.05) were significantly increased in the tensile stress group; The mRNA expression of matrix proteinase ADAMTS3 ( t=5.241, P<0.05) and MMP19 ( t=24.72, P<0.01) were significantly decreased, and TIMP3 ( t=8.472, P<0.01) increased in the tensile stress group; The mRNA expression of extracellular matrix molecules COL2A1 ( t=5.871, P<0.05), FLRT2 ( t=5.216, P<0.05) and FN1 ( t=4.289, P<0.05) were significantly increased. Conclusion:The cyclic tensile stress promoted cell cycle and proliferation and inhibited apoptosis of nucleus pulposus cells. The cyclic tensile stress may affect the function and degeneration of nucleus pulposus cells by regulating the expression of inflammatory related factors, TGF-β, matrix degradation enzymes and ECM molecules.

Objective: To review the systematic reviews of acupuncture for diabetic peripheral neuropathy (DPN) and to provide evidence for clinical decisions. Methods: Published systematic reviews targeting acupuncture treatment of DPN were searched using computer through both Chinese and English databases till July 1, 2019. Two researchers screened the papers based on inclusion and exclusion criteria and conducted report quality evaluation, methodological quality assessment and evidence quality grading using the preferred reporting items for systematic reviews and meta-analyses (PRISMA), assessment of multiple systematic review 2 (AMSTAR 2) and grading of recommendations assessment, development and evaluation (GRADE). Results: Ten systematic reviews were included, involving 11 outcome measures. According to PRISMA, 6 items were sufficiently reported while 1 item was not; AMSTAR 2 appraised that all the included systematic reviews were of low quality in the methodological evaluation; according to GRADE, of the 30 clinical evidences, only 5 were graded moderate while the remained were graded low or extremely low. Descriptive analysis showed that acupuncture can significantly improve DPN symptoms, accelerate the conduction velocities of sensory and motor nerves, and up-regulate the content of plasma nitric oxide (NO), while the adverse reaction rate was low. Conclusion: Acupuncture can produce satisfactory clinical efficacy in treating DPN, but the existing problems, such as low-quality evidence, unitary outcome measures, poor methodological quality of systematic reviews and nonstandard reporting, need to be treated cautiously; meanwhile, more high-quality clinical trials are required to elevate the level of evidence.

Objective To determine the activities of 131I for treating differentiated thyroid carcinoma with diffuse pulmonary metastases ( DTC-DPM ) from the perspective of internal radiation dosimetry.Methods According to Medical Internal Radiation Dosimetry (MIRD) schema, the activity constraint,from which the whole bdy retention at 48 h should not exceed 2.96 GBq (2.96 GBq rule), was converted to dose-rate constraint(DRC) to lungs at 48 h ( DRCLU ·48 h ) in 131I therapy for DTC-DPM. Based on the assumption of DRCLU·48 h at 48 h in lung, the fractions of whole body activities ( F48 ), the effective half times of 131I in lungs ( TLL ) and the remainder of body ( TRB ) were 0.6-0.9, 20- 120 h, and 10- 20 h, respectively. The maximum safe activities of 131I for different human phantoms from the Organ Level Internal Dose Assessment (OLINDA) software were calculated. Results According to MIRD schema and 2.96 GBq rule, DRCLU ·48 h should not exceed 46.4 mGy/h in 131I therapy for DTC-DPM. Depending on varying F48 h,TLL and TRB, the maximum safe activities of 131I were 6.77-81.36, 5.29-56.20, 5.08-55.19 and 3.87-40. 52 GBq for the male adult, female adult, 15-year-old, and 10-year-old patients with DTC-DPM, respec tively. Conclusion Dosimetry-guided 131I therapy for DTC-DPM considers adequately the differences of 131I kinetics in individual patients and can adjust administered activities of 131I on the precondition of avoiding radiological pneumonitis and pulmonary fibrosis.

ObjectiveTo investigate the feasibility and optimal condition of isolation,purification and expansion of mesenchymal stem cells(MSCs) derived from human umbilical cord blood in vtro.MethodsHuman umbilical cord blood(HUCB) was collected from full term deliveries scheduled,all samples were obtained sterilely with 20 U/ml preservative free heparin.The cord mononuclear cells were isolated with lymphocyte separation medium(density 1.077 g/ml),purified and expanded with MesencultTM medium and acidic environment to produce adherent layer.The surface antigen expression of MSCs was detected with flow cytometry.ResultsThe HUCB-derived mononuclear cells,when seeded in specific medium,gave rise to adherent cells,which exhibited either an osteoclast or mesenchymal-like phenotype.After passage 3,these cells were able to be purified and expanded.6.6×105 primary MSCs reached a number of 9.9×10<>sup8 after 10 expanded passage.Flow cytometry showed that MSCs did not express antigens CD34,CD11a and CD11b,but express strongly CD29 and weakly CD71,which was identical to human bone marrow-derived MSCs.ConclusionMSCs in HUCB can be cultured and expanded in vitro,and could be a source of stem cells for experimental and clinical application.

Objective To summarize the clinical experience of standard piggyback liver transplantation(SPBLT) and modified piggy-back liver transplantation(MPBLT) in the treatment of Wilson′s disease(WD).Methods The clinical records of 29 cases of WD who underwent piggyback liver transplantation over the recent 12 years were analyzed retrospestively.Among them,there were 22 male and 7 female patients,with 6cases aged 8-14y and 23 cases aged 24-37y,who underwent liver transplantation because of chronic advanced liver disease(18cases),fulminant hepatic failure(2 cases),or with normal liver function(9 case),and among them,24 cases showed neurological dysfunction.The modes of operation included SPBLT(13 cases) and MPBLT(16 cases).Results The patients were followed up for 1 month-12 years with the median follow-up time of 47 months.Four patients died in the perioperative period,of which 2 died of hepatic failure due to obstruction of hepatic venous return after SPBLT,and 2 died of hepatic artery thrombosis and bile leakage combined with severe infection.The recipient survival rate at 1 year and 3 years was 86% and 79% respectively.One patient has survived for 12 years.All the levels of serum copper and copper-protein recovered to normal in four weeks post-operatively.Neurological symptoms improved in varying degrees after operation.Conclusions WD patients who undergo PBLT can expect a satisfactory clinical outcome and a good quality of life,and MPBLT can effectively reduce the complications of operation and improve the operation survival rate.