Traditional Chinese medicine(TCM)is the most original innovation science. The innovation and development of TCM is based on the precious treatment theory and copious clinic experience of predecessor. Combination of disease and syndrome is the important approach of innovation and development of traditional Chinese medicine.Establish of a perfect evaluation system of clinical effect which has special feature of TCM is the key of innovation and development of traditional Chinese medicine.

To construct a system of genetic transformation suitable for Rhizopus oryzae, we constructed a single-exchange vector pBS-hygro carrying hygromycin B resistance gene (hph) as its selective marker using gene splicing by overlap extension PCR (SOE PCR) technique. We introduced this recombinant vector into Rhizopus oryzae AS 3.819 by PEG/CaCl2-mediated transformation of protoplast, electroporation of protoplast and germinated spores; and we studied the effects of hydrolysis time, field strength and spore germination time on transformation frequency. We conducted quantitative real-time PCR (qPCR) assay to determine the gene copy number of ldhA integrated in the genome of R. oryzae transformants and its effect on the stability of transformants. We successfully achieved R. oryzae transformants integrated with pBS-hygro-ldhA vector. The optimal hydrolysis time for protoplast production was 140 min, and the optimal field strength of electroporation pulse for protoplast was 13 kV/cm. The optimal germination time of spores for electroporation was 2.5 h, and the optimal field strength of electroporation pulse was 14 kV/cm. The transformation frequency of method based on germinated spores was generally higher than the methods based on protoplast. The qPCR test results suggested that transformants with high copy number of integration in a certain range were relatively stable. Our results provided basis and support for metabolic regulation and genetic engineering breeding of R. oryzae.
DNA, Recombinant ; Electroporation ; Genetic Engineering ; Genetic Vectors ; Hygromycin B ; Protoplasts ; Real-Time Polymerase Chain Reaction ; Rhizopus ; genetics ; Transformation, Genetic

OBJECTIVE: To explore the influence of the interaction among occupational stress, sleep duration and sleep quality on the prevalence of hypertension in petroleum workers. METHODS: A total of 3 040 workers from six oil field bases in Karamay City were selected as study subjects by multi-stage random cluster sampling method. The Chinese version of Pittsburgh Sleep Quality Scale and the revised version of Occupational Stress Scale were used to evaluate their sleep quality and occupational stress status. Binary logistic regression was used to analyze the effect of interaction of occupational stress, sleep duration and sleep quality on hypertension. RESULTS: The prevalence of hypertension in the study subjects was 15.3%(466/3 040), and the detection rates of sleep deprivation, poor sleep quality and high occupational stress were 26.5%, 78.3% and 19.6% respectively. After adjusting for confounding factors such as gender, ethnicity, age, marital status, education level, length of service, professional title, shift work, smoking, alcohol consumption and body mass index, the interaction analysis results showed that the risk of hypertension was higher in the poor sleep quality groups with normal sleep duration, sleep deprivation or longer sleep duration than that in good sleep quality group with normal sleep duration(all P<0.05), respectively. The risk of hypertension was higher in the group with sleep deprivation and high occupational stress than that in the group with normal sleep duration and low occupational stress(P<0.01). In the group with poor sleep quality and high occupational stress the risk of hypertension was higher than that in the group with good sleep quality and low occupational stress(P<0.05). CONCLUSION: The interaction among occupational stress, sleep duration and sleep quality may increase the risk of hypertension in petroleum workers.

OBJECTIVETo observe the efficacy of chemotherapy consisted of bortezomib as main druy in maintenance therapy for recurrence of newly diagnosed MM patients.

METHODSThe clinical data and outcome of 37 MM patients during 2008-2013 were analyzed retrospectively, the 37 MM patients were divided into 2 group: 19 cases including 13 cases of newly diagnosed MM with symptoms and 6 cases of relapsed refractory MM were enrolled in group A; 17 cases of newly diagnosed MM with symptoms were enrolled in group B. The patients of group A received maintenance therapy consisted of bortezomib plus dexamethasone (VD group), while the patient group B received maintenance therapy consisted of melphalan plus prednisone(MP group), then the therapeutic efficacy of 2 group was compared.

RESULTSThe overall response rate(ORR) in VD groupe was 84.2%(16/19), out of which CR rate reached 42%(8/19), PR rate reached 31.6%(6/19), MR rate reached 10.5%(3/19). During median follow-up for 21.8(5-51) months, death occurred, while the ORR in MP group was 52(9/17), out of which CR rate was 23.5%(4/17), PR rate reached 23.5%(4/17), MR rate reached 5.9%(1/17). Druing median follow-up for 16.4(4-39) months, the worteity reaced 64.7%(11/17). The differencr between 2 groups was significant(P<0.05). The median OS time of patients in VD group was 21.6 months, that in MP group was 17.9 months(P<0.05). The median PFS in VD group and MP group were 13.4 and 9.4 months respectively(P<0.001).

CONCLUSIONThe ORR and CR rates of bortezomib maintenance therapy for newly diagnosed and relapsed / refractory MM patients are very high, and its toxicity can be controlled, therefore, the patients need maintenance therapy after remission.

Objective To measure the accuracy of multi-leaves collimator (MLC) leaves positions in intensity modulated radiation therapy (IMRT) for verification purpose.Methods Solid water homogeneous phantom with size of 30 cm× 30 cm was scanned by CT scanner.The scanned images were delivered to radiation therapy plan system (TPS) to formulate the therapy plan.The MLC leaves created 5 strips of exposure field,each 3 cm long and 0.6 cm wide.The strip-to-strip distance was 3 cm.With 6 MV X-rays,the SSD was 100 cm at the maximum dose point.The MU per strip was 250 MU.EBT2 radiochromic films each of 25 cm×25 cm were placed on the 30 cm×30 cm homogeneous solid phantom and covered with 1 cm thick solid phantom slabs for delivering of IMRT.Results For 7 of 8 accelerators,the differences of film-measured and TPS-planned MLC leaf position for every fence field were within ± 0.5 mm as required by IAEA,with only other one being-0.6 mm,not consistent with the IAEA requirements.The film-measured position difference between each pair and all pairs of leaves for 8 accelerators were all in line with IAEA's requirements.The film-measured actual width difference between each pair and all pairs of leaves was within ±0.75 mm as required by IAEA for 4 accelerators and outside ±0.75 mm for the other three,not consistent with the IAEA requirements.The standard deviation of film-measured actual width between each pair and all pairs of leaves for 6 accelerators were ≤ 0.3 mm,as required by IAEA,whereas ＞0.3 mm for the other two,not consistent with IAEA requirements.Conclusions The film dosimetric verification of IMRT is an important part of its quality assurance,featuring simple,reliable and highly accurate positioning and can meet measurement requirement.

OBJECTIVETo investigate the correlation of patients with thrombosis or prothrombotic status with hyperhomocysteinemia (HHcy), activated protein C-resistance(APCR) and gene polymorphism of coagulation factor V.

METHODSThree hundred healthy voluteers were selected as controls, 223 cases of thrombosis (80 cases of cerebral infarction of CT, the MI of 82 cases of myocardial infarction, venous thrombosis of VTE 61 cases), 270 cases of patients with prothrombotic state (76 cases of pregnancy disease of PIH, 62 cases of chronic obstructive pulmonary disease (COPD), 60 cases of diabetes(DM) and 72 cases of cancer) were enrolled in this study. The plasma APCR and hyperhomocysteinemia were detected by APTT coagulation method and cycling enzyme method respectively, and restriction fragment length polymorphism(RFLP) were was used to detect the gene polymorphism of FV G1691-A, G1091-C and A1090-G in the patient and control groups.

RESULTSAPCR positive rate was 62.29% and 7.33%, and the positive hyperhomocysteinemia accounted for 68.42% and 10.00% respectively in the group of the patients with venous thrombosis and the normal control group. 3 cases of heterozygous FV gene mutations were found in the APCR-positive patients with venous thrombosis.

CONCLUSIONHHcy possitive rate of patients with venous thrombosis is signiticantly higher than that in control, the HHcy is one of the important causes resulting in thrombosis, the patients with venous thrombosis have proved to be with APCR, and the possitive APCR may be related with the coagulation factor V gene polymorphism.

Acetyl dipeptide-1 cetyl ester (AD-1) is a synthetic peptide composed of acetic acid and cetyl alcohol with arginine and tyrosine, which has certain anti-inflammatory and skin barrier enhancement effects, has been used in cosmetics for sensitive skin.Meanwhile, the ingredient has also been used in anti-aging cosmetics, but there is a lack of published scientific evidence on anti- senescence aspect.In this study, we investigated the related effects of AD-1 by evaluating its in vitro antioxidant and antiglycation efficacies.Furthermore, we established a photoaging model on primary rat dermal fibroblasts by repeated exposures to UVA irradiation.MTT assay was used to detect the effects of AD-1 on the cell viability.RT-qPCR was used to determine the effects of AD-1 on the mRNA levels of senescence-related p21, p53, MMPs, IL6, Col1, Col3 and autophagy-related p62, ATG5, ATG7.Western blot was used to detect the effects of AD-1 on the protein levels of p16, p21, p53, Col1, LC3B and p62.SA-β-gal was performed to indicate senescence level of the cell.MDC was performed to indicate autophagy level.Intracellular reactive oxygen species were monitored by fluorescent probes DCFH-DA.The results showed that AD-1 could reduce UVA-induced the cell damage and regulate the abnormal expression of mRNA levels. It alleviated the abnormal protein levels of p16, p21, p53, Col1, LC3B and p62 induced by UVA. These results suggested that AD-1 has not only antioxidant and antiglycation effects but also can activate autophagy to achieve anti-senescence effect.

The aim of this study was to produce recombinant porcine interferon gamma (rPoIFN-γ) by Chinese hamster ovarian (CHO) cells expression system and to analyze its antiviral activity. Firstly, we constructed the recombinant eukaryotic expression plasmid pcDNA3.1-PoIFN-γ and transfected into suspension cultured CHO cells for secretory expression of rPoIFN-γ. The rPoIFN-γ was purified by affinity chromatography and identified with SDS-PAGE and Western blotting. Subsequently, the cytotoxicity of rPoIFN-γ was analyzed by CCK-8 test, and the antiviral activity of rPoIFN-γ was evaluated using standard procedures in VSV/PK-15 (virus/cell) test system. Finally the anti-Seneca virus A (SVA) of rPoIFN-γ activity and the induction of interferon-stimulated genes (ISGs) and cytokines were also analyzed. The results showed that rPoIFN-γ could successfully expressed in the supernatant of CHO cells. CCK-8 assays indicated that rPoIFN-γ did not show cytotoxicity on IBRS-2 cells. The biological activity of rPoIFN-γ was 5.59×10⁷ U/mg in VSV/PK-15 system. Moreover, rPoIFN-γ could induced the expression of ISGs and cytokines, and significantly inhibited the replication of SVA. In conclusion, the high activity of rPoIFN-γ was successfully prepared by CHO cells expression system, which showed strong antiviral activity on SVA. This study may facilitate the investigation of rPoIFN-γ function and the development of novel genetically engineered antiviral drugs.
Swine ; Animals ; Cricetinae ; Interferon-gamma/pharmacology* ; Cricetulus ; CHO Cells ; Sincalide ; Recombinant Proteins/pharmacology* ; Antiviral Agents/pharmacology*