Fungal polyketide synthases are responsible for the biosynthesis of secondary metabolites such as pigments, mycotoxins, and they are very important in pharmacology , food science and agriculture. The recent advances in the methods for the isolation and manipulation of multiple classes of polyketide synthase genes from fungi were introduced. It is useful for discovery of novel fungal polyketide synthase gene clusters. These methods can also be useful for revealing the genetic potential of fungi to produce multiple types of bioactive polyketide.

Aim To construct the cell model targeted on the damage by α-synuclein for screening anti-Parkinson’s Disease (PD)compounds.Methods The cDNA fragment of α-synucle-in gene was obtained by PCR methods and inserted into the re-combinant prokaryotic plasmid by molecular cloning technique. The recombinant plasmid was transformed into Escherichia coli, and subsequently induced to express α-synuclein protein.The recombinant α-synuclein was purified and identified by affinity chromatography,immunoblotting and mass spectrometry.The cells damage by α-synuclein was evaluated through cell viability measured by 3-(4,5-dimethyl-2-thiazolyl )-2,5-diphenyl-2-H-tetrazolium bromide.Results The obtained cDNA fragment ofα-synuclein in accordance with its theoretic molecular weight was cloned into pET30a plasmid and verified by sequencing.The re-combinant plasmid was transformed into bacteria E.Coli.BL21 (DE3)and induced to express α-synuclein by isopropyl β-D-1 -thiogalactopyranoside (IPTG).The expression condition was op-timized according to the culture temperature,the concentration of IPTG and the proliferation state of bacteria.The purified α-synu-clein was proved to be a 1 5.3 ku molecule weight protein,and could be immunoblotted with anti-α-synuclein antibody.The pu-rified α-synuclein could decrease the viability of PC1 2 cells and primary neurons significantly,and its effect was in a concentra-tion-dependent manner.Conclusion We have succeeded in constructing the cell model targeted on the damage by α-synucle-in.

BACKGROUND: The extramedulary fixation system including dynamic hip screw (DHS) is commonly used in treatment of Intertrochanteric fracture. However, in patients with unstable intertrochanteric fracture, extramedulary fixation system often leads to the failure of fracture fixation. Intramedulary fixation system including both proximal femoral nail antirotation (PFNA) and InterTan nail has been widely used in the treatment of unstable intertrochanteric fractures. OBJECTIVE:To compare the therapeutic effects of extramedulary fixation system containing DHS, PFNA and InterTan nail in the treatment of intertrochanteric fracture. METHODS:Literatures were searched in Wanfang, PubMed, Embase, Medline, the Cochrane library to screen literatures published from January 1990 to November 2014. Relevant studies addressing extramedulary fixation system containing DHS, PFNA and InterTan nail were screened. RESULTS AND CONCLUSION: 346 articles were screened, and 13 of them were in accordance with the inclusion criteria. 1 271 patients with different types of intertrochanteric fracture were assessed in this study. Compared to DHS group, patients treated with PFNA and InterTan nail had shorter operation time and less blood loss. No significant difference in rehabilitation time and Harris score was detected among three kinds of fixation methods. Additionaly, PFNA and InterTan nail had a similar effect. These findings verify that compared with DHS, PFNA and InterTan nail can optimize the surgery, but cannot elevate postoperative outcomes.

Cytokine-Induced Killer Cells ; immunology ; Cytotoxicity, Immunologic ; Dendritic Cells ; immunology ; Fetal Blood ; immunology ; HL-60 Cells ; Humans ; K562 Cells ; Killer Cells, Natural ; immunology

This study examined the current changes of human ether-a-go-go-related gene (hERG) mutation derived from a LQT2 Chinese family with a highly penetrating phenotype. Mutation was identified and site-directed mutagenesis was performed to induce the mutation in wild-type (WT) hERG. WT hERG and mutated V535M were cloned and transiently expressed in HEK293 cells. At the 48th and 72nd h after transfection, membrane currents were recorded using whole cell patch-clamp procedures. An A>G transition at 1605 resulting in replacement of V535M was identified. Compared to WT, V535M mutation significantly decreased tail currents of hERG. At test potential of -40 mV after depolarizing at +50 mV, tail current densities were 83.35±7.06 pA/pF in WT and 50.38±7.74 pA/pF in V535M respectively (n=20, P<0.01). Gating kinetics of hERG revealed that V (1/2) of steady-state inactivation shifted to negative potential in the mutant (V (1/2,V535M): -61.81±1.7 mV vs. V (1/2, WT): -43.1±0.71 mV). The time constant of recovery from inactivation was markedly prolonged in the mutant compared to WT among test potentials. V535M hERG mutation demonstrated markedly decreased tail current densities, which suggests that V535M is a new loss-of-function mutation of hERG channel responsible for LQT2.

The paper is focused on the clinical applications of Tanreqing injection after listing, detecting and analyzing the related blood indicators of patients with allergic reactions based on prospective, multi-center, large sample, registration-type clinical safety monitoring nested case-control study (NCCS) to explore the possible mechanisms of allergic reaction of Tanreqing injection, 3 006 patients cases used with Tanreqing injection were monitored, including 3 cases of adverse reactions and 2 cases of allergic reactions. Each patient of allergic reactions, according to the ratio of 1:4 matches four cases of not adverse reactions as a control group of patients, while 5 healthy and 5 cases of volunteers into the healthy group. We examined the correlation detection of cases of allergic reactions among groups such as T-IgE, IgA, IgG, C3, C4, IFN-g, IL-2, IL-4, IL-6, IL-10. Allergic reactions of Tanreqing injection may be mediated by IgE as type I based on existing research data. This results and conclusions will promote the justifiability and safety of clinical applications.
Adolescent ; Adult ; Case-Control Studies ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; Female ; Humans ; Hypersensitivity ; diagnosis ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-2 ; metabolism ; Interleukin-4 ; metabolism ; Interleukin-6 ; metabolism ; Male ; Prospective Studies ; Young Adult

Objective To isolate Fusarium species from Kashin-Beck disease(KBD)area and biosynthesize cnlde T-2 toxin.Methods T-2 toxin.producing Fusarium was isolated from corns produced in KBD area and purifted.The purifted funsi were identified according to the traits of colony,appearance of thallus and characters of conidium and then weIe cultivated in sterile Corn culture media.After extraction with organic solvent and purification by silica gel chromatography column,the quality and quantity of the toxin in the extracts were estimated by thin,Layer chromatography and high-performance liquid chromatography.Results The toxin-producing strain was Fusarium tricinctum. The com cuIture media inoculated with this strain produced about 250 mg of crude T-2 toxin per kg. Conclusions This experiment has indirectly further confirmed pollution of T-2 toxin-producing Fmarium existed in

Objective To investigate the prevalence of vitamin D deficiency and its associated metabolic risk factors in the healthy adults in Gansu Province, a western province of China.Methods Through stratified, multistage probability population sampling, 11 300 adults (4 740 men and 6 417 women aged 18-79 years) who had been living in their current residence for at least 5 years were arbitrarily chosen from seven representative geographical cities in Gansu Province from June 2014 to December 2015.The data obtained from a questionnaire-based survey, anthropometric and biochemical assessments, and areal bone mineral density measurement were collected and analyzed.Results The overall mean serum 25(OH)D was (16.07±9.40) μg/L.The prevalence of severe deficiency (<10 μg/L), deficiency (≥10 μg/L and <20 μg/L), insufficiency (≥20 μg/Land <30 μg/L), and sufficiency (≥30 μg/L) among these adults were 17.3％, 64.6％, 11.8％, and 6.3％, respectively.The significant predictors of vitamin D deficiency included younger age and female sex(P<0.05), whereas sun exposure, physical activity, and calcium supplementation were protective (P<0.05).Serum 25(OH)D was inversely associated with parathyroid hormone (r=-0.279, P<0.001) and positively associated with serum calcium (r=0.239, P<0.001), serum phosphorus (r=0.090, P=0.018), low-density lipoprotein cholesterol(r=0.100, P=0.008), and body mass index (r=0.093, P=0.014).The 25(OH)D concentration showed no significant correlation with bone mineral density (P>0.05).Conclusions Vitamin D deficiency is highly prevalent among adult populations in Gansu, northwestern of China, and is largely attributed to younger age and female sex.The cumulative effects of factors including sun exposure, physical activity, and calcium supplementation play an influential role in maintaining vitamin D levels.

Objective To evaluate the safety and rationality of total/near total bilateral thyroidectomy(TBT) for patients with bilateral multinodular goiter(BMG). Methods From January 2003 to December 2006,311 BMG cases were preoperatively divided into two groups, 130 cases in group A underwent TBT, and 181 cases in group B were treated with subtotal/partial bilateral thyroidectomy. Results There were 6 and 2 eases in group A and group B respectively diagnosed by intraoperative frozen biopsy as BMG, but identified as papillary carcinoma by final pathology. Hence the 6 cases in group A avoided reoporation, while the 2 cases in group B underwent a resection of the remnant gland. Transient hoarseness developed in 3 (2.42%, 3/124) and 3 (1.68%, 3/179) eases in group A and group B respectively (P =0.48). Transient hypocalcemia developed in 11 (8.87% ,11/124) and 9(5.03% ,9/179) cases in group A and group B respectively(P =0.16). There was no postoperative goiter recurrence in group A, but recurrence developed in 12 cases (6.70%,12/179) in group B(P=0.02). Conclusions Total bilateral thyroidectomy is safe and rational for the management of bilateral thyroid goiter.

BACKGROUND: The growth behaviors of cells on the biomaterials scaffold may be affected by the topography, pore size, wettability, porosity and other factors.OBJECTIVE: This research is aimed to observe the growth and proliferation of Schwann cells in silk fibroin scaffolds with different pore sizes. METHODS: Two kinds of silk fibroin scaffolds with different pore sizes were prepared, including a large pore size scaffold (pore size 50-60 μm) and a small pore size scaffold (pore size 10-20 μm). Schwann cells (R3 [33-10ras3]) served as seed cells and incubated in 37 ℃, 5% CO2 incubation box. When cells filled up the culture bottle bottom and formed a dense monolayer, they were digested and the cell concentration adjusted, then Schwann cells were seeded onto the surface of the porous silk fibroin scaffolds with different pore sizes. After seven days of co-culture, the growth and proliferation of Schwann cells were observed under scanning electron microscope.RESULTS AND CONCLUSION: The growth of Schwann cells on the surface of silk fibroin scaffolds with different pore sizes was varied. On the surface of small pore size scaffold (10-20 μm), the cell density was low, while the phenotype of cells was bipolar, cells arranged in parallel or linked as the cell chains. On the surface of large pore size scaffold (50-60 μm), more cells could be seen, but most of the cells were in the shape of single sphere, cells clustered on the surface of the porous scaffold or aggregated as a bunch of grape at the bottom of pores. Only few cells were bipolar and lied on the ridge between the pores. The result showed that the pore size of porous silk fibroin scaffolds is an influential factor for the growth and adhesion of Schwann cells. Schwann cells are conducive to grow on the scaffolds with pore size larger than cell body diameter.