Cloning and protein expression analysis of monoterpene synthase gene TwMS in Tripterygium wilfordii / 中国中药杂志

In this study, we cloned a monoterpene synthases, TwMS from Tripterygium wilfordii suspension cells. TwMS gene contained a 1 797 bp open reading frame (ORF), encoding a polypeptide of 579 amino acids, which deduced isoelectric point (pI) was 6.10 and the calculated molecular weight was 69.75 kDa. Bioinformation analysis showed that the sequence of TwMS was consistent with the feature of monoterpene synthases. Differential expression analysis revealed that the relative expression level of TwMS increased significantly after being induced by methyl jasmonate (MeJA). The highest expression level occurred at 24 h. TwMS protein was successfully expressed in Escherichia coli BL21 (DE3), which laid the foundation for identifying the function of T. wilfordii monoterpene synthases.

Cloning and expression analysis of kaurenoic acid oxidase gene in Tripterygium wilfordii / 中国中药杂志

Kaurenoic acid oxidase involved in biosynthesis pathway of gibberellin. According to the transcriptome database, the specific primers were designed and used in cloning the full-length cDNA of TwKAO, the bioinformatic analysis of the sequence was performed. The qRT-PCR were used to detect the expression level of TwKAO after MeJA treatment.The full-length cDNA of the TwKAO was 1 874 bp encoding a polypeptide of 487 amino acids.The calculate molecular weight was about 56.02 kDa,and the theoretical isoelectric point (pI) was 8.89. The relative expression level of TwKAO was deduced by MeJA and reached the highest at 12 h after the treatment.Plant tissue expression analysis indicated that, TwKAO expressed the highest in leaves,while lowest in roots.For the first time, we cloned and analyzed the expression characteristics of TwKAO, which laid a foundation for deep analysis of growing development and terpenoid secondary metabolites in T. wilfordii.

Cloning and protein expression analysis of geranyl diphosphate synthase genes in Tripterygium wilfordii / 中国中药杂志

Based on the transcriptome data, the study cloned full-length cDNA of TwGPPS1 and TwGPPS2 genes from Tripterygium wilfordii suspension cells and then analyzed the bioinformation of the sequence and protein expression. The cloned TwGPPS1 has a 1 278 bp open reading frame (ORF) encoding a polypeptide of 425 amino acids. The deduced isoelectric point (pI) was 6.68, a calculated molecular weight was about 47.189 kDa. The full-length cDNA of the TwGPPS2 contains a 1 269 bp open reading frame (ORF) encoding a polypeptide of 422 amino acids. The deduced isoelectric point (pI) was 6.71, a calculated molecular weight was about 46.774 kDa.The entire reading frame of TwGPPS1,2 was cloned into the pET-32a(+) vector and expressed in E. coli BL21 (DE3) cells to obtain the TwGPPS protein, which laid a basis for further study on the regulation of terpenoid secondary metabolism and biological synthesis.

Cloning and protein expression of sterol-C-24-methyl transferase 2 in Tripterygium wilfordii / 药学学报

24-Alkyl sterols are the major players in the control of membrane component and plant growth. In this paper, we cloned an important rate-limiting enzyme:sterol-C-24-methyl transferase (SMT) in the sterol biosynthetic pathway according to the transcriptome data of Tripterygium wilfordii. suspension cells, whose full-length cDNA was 1 631 bp with an open reading frame of 1 080 bp, encoding a protein of 359 amino acids. It was estimated that theoretical isoelectric point (pI) was 6.43 and the molecular mass was 40.0 kDa. Bioinformatics analysis attributed the SMT gene to SMT2 family. The expression vector was constructed as the pMAL-c2x-TwSMT2 plasmid and the recombinant protein was expressed in E. coil BL21 (DE3) competent cells. After methyl jasmonate treatment, the relative expression level of TwSMT2 has improved significantly in 24 h. SDS-PAGE electrophoresis and Western Blot showed that protein of TwSMT2 in BL21 (DE3) strain was expressed after induction by IPTG. In this study, TwSMT2 was cloned for the first time and the recombinant protein was expressed, which lay the foundation for elucidation of the sterol biosynthetic pathway of Tripterygium wilfordii in the future.

The study of the characteristics and influencing factors of pneumoconiosis among workers exposed to dusts in an iron mine / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To analyze the prevalence characteristics and influence factors of pneumoconiosis of workers exposed to dusts in an iron mine, to provide the base of preventive measures for pneumoconiosis of iron mine.</p><p><b>METHODS</b>The subjects of cohort study were all workers exposed to dusts for at least one year registered in an iron mine during 1960 to 1974, and followed-up to the end in 2003. The cases with pneumoconiosis were diagnosed by the local diagnosis group of pneumoconiosis, according to the national diagnostic criteria of pneumoconiosis. The risk factors were analyzed with Cox risk model.</p><p><b>RESULTS</b>A total of 3647 miners were included in the cohort study and were followed up by 132 574.4 person years. There were 316 cases with pneumoconiosis, and the incidence of pneumoconiosis for a year was 0.24 per thousand. There were 274 cases (86.7%) with pneumoconiosis in workers exposed to dusts before 1960, the incidence of pneumoconiosis for a year was 0.40 per thousand, which was significantly higher than that (0.07 per thousand) of workers exposed to dusts after 1960. The average latency of pneumoconiosis was 26.0 +/- 7.3 years. The average durations of upgrade from stage 0(+) to I , I to II and II to III were 5.3 +/- 3.2, 6.6 +/- 5.2 and 11.3 +/- 5.0 years, respectively. However, 164 cases with pneumoconiosis were diagnosed after ceasing exposure to dusts for mean 8.3 years. The risk of pneumoconiosis in iron miners increased with exposure doses, and there was an obvious dose-effect relationship. The average cumulative exposure dose of cases with pneumoconiosis was 173.7 +/- 91.6 mg/m3 x y, which was significantly higher than that (112.1 +/- 64.8 mg/m3 x y) of workers without pneumoconiosis. Also the tuberculosis (HR = 5.9, P < 0.001) and smoking (HR = 1.7, P < 0.01) were the main risk factors.</p><p><b>CONCLUSION</b>There was an obvious dose-effect relationship between the cumulative exposure dose and pneumoconiosis incidence. Tuberculosis and smoking were the main risk factors influencing the pneumoconiosis incidence.</p>

Preparation of polyvinyl alcohol/chitosan hybrid nanofibrous scaffolds and study on its biodegradation behaviors in vivo and in vitro / 中华烧伤杂志

<p><b>OBJECTIVE</b>To fabricate polyvinyl alcohol (PVA)/chitosan hybrid nanofibrous scaffolds owning the similar physiological structure of ECM, and to observe its biodegradation behavior in vivo and in vitro.</p><p><b>METHODS</b>(1) The PVA nanofibrous scaffold and PVA/chitosan hybrid nanofibrous scaffold were fabricated by electrospinning technique, and then they were crosslinked by glutaraldehyde vapor method. The morphology of both scaffolds was observed by scanning electron microscope (SEM). (2) Biodegradation experiment in vitro: the samples of two scaffolds with size of 2 cm x 2 cm were placed into phosphate-buffer saline (PBS) fluid under 37.0 degrees C water for incubation, and then they were dried to observe morphologic changes under SEM on post incubation day (PID) 3, 7, and 14. (3) Biodegradation experiment in vivo: 48 Wistar rats were divided into PVA group and PVA/chitosan group according to the random number table, with 24 rats in each group. PVA or PVA/chitosan nanofibrous scaffold was implanted into subcutaneous tissue on both sides of back in rats of both groups, with 4 scaffolds in each rat. The scaffold samples were harvested to observe morphologic changes with HE staining on post operation day (POD) 3, 7, 14, and 28.</p><p><b>RESULTS</b>(1) After crosslinking, the surface of fibers in PVA and PVA/chitosan hybrid nanofibrous scaffolds were smooth, and the diameters of fibers were similar, ranging from 200 to 300 nm, with high porosity. (2) Biodegradation experiment in vitro showed that the morphologic changes in fiber was respectively swelling, dissolution, fusion in PVA nanofibrous scaffold on PID 3, 7, 14, and that in PVA/chitosan hybrid nanofibrous scaffold was respectively swelling, dissolution and fragmentation, and disappearance. (3) Biodegradation experiment in vivo showed that the morphologic changes in scaffold structure was respectively loosening, fuzziness of edges, degradation, and disappearance in PVA group and PVA/chitosan group on POD 3, 7, 14, 28.</p><p><b>CONCLUSIONS</b>PVA/chitosan hybrid nanofibrous scaffolds can be prepared with electrospinning technique, and it has an appropriate biodegradation rate compatible with tissue reconstruction after crosslinking.</p>

Polymorphism of TPH gene T3792A locus in Han ethnic group of north China / 法医学杂志

OBJECTIVE@#To investigate the polymorphism of the TPH gene T3792A locus in Han ethnic group of north China and its application value in forensic science.@*METHODS@#The polymorphism of T3792A locus of the TPH gene was analyzed by the ASPCR of blood samples from 173 unrelated individuals of north Chinese Han population.@*RESULTS@#The distribution of the T3792A locus polymorphism of the TPH gene in Han ethnic group of north China followed the Hardy-Weinberg law, with the allele A and T gene frequency of 0.486 and 0.514, respectively.@*CONCLUSION@#The TPH gene T3792A locus shows a very good genetic polymorphism, and may be applied to individual identification and paternity testing.

Investigation on mental health of residents living in a mineral area in Hubei province / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To study the effect of mining operations on the mental health of residents living in a mining area in Hubei province.</p><p><b>METHODS</b>Villagers (n = 93) living in the mining area were compared with a control group of residents (n = 101) in mental health status, and knowledge of environment and health. The mental health status of villagers was assessed using the Symptom Checklist 90 (SCL-90), State-Trait Anxiety Inventory (STAI); The knowledge of environment and health was evaluated using a self-designed questionnaire. The urine and hair samples were collected from some subjects. The lead, cadmium, arsenic, copper and zinc contents were detected as well as the total protein, NAG, d-ALA in the urine.</p><p><b>RESULTS</b>The occurrence rate of lead, cadmium, arsenic exposure symptoms was significantly higher in the exposed group than in the control group. The urine cadmium, the hair cadmium, the hair arsenic and hair lead were significantly higher in the exposed group than in the control group (P < 0.05). The positive symptom detection rate of SCL-90 in the exposed group was 8.60% compared with 0.99% of the control group. For the SCL-90, the total scale, somatization, obsessive-compulsive, interpersonal sensitivity, depression, anxiety, hostility, paranoid ideation, psychoticism, other symptom in the exposed groups were significantly higher than those in the control group (P < 0.05), indicating the status of the psychological hygiene of the exposed group was worse than the control group. The total S-AI (anxiety) score and the sex-specific value in the exposed group were higher than the control group (P < 0.05, P < 0.01), which showed that the anxiety of the exposed group was more evident. The total T-AI score and the sex-specific value in women of the exposed group were significantly higher than the control group (P < 0.01), showing that the anxiety were more significant in the exposed group, especially the women of the exposed group.</p><p><b>CONCLUSION</b>The mental health status of the residents who living in a mining area is affected and they have a higher S-AI and T-AI scale than those living a non-mining area.</p>

Distribution of tryptophan hydroxylase gene-T457C locus polymorphism in Han ethnic group in northern China / 法医学杂志

OBJECTIVE@#To study the distribution of the tryptophan hydroxylase (TPH) gene-T457C locus polymorphism in Han ethnic group in northern China and to find its applicable value in forensic science.@*METHODS@#Genomic DNA samples, extracted from 180 unrelated individuals in northern Chinese Han population, were analyzed by PCR-RFLP.@*RESULTS@#The discrimination power (DP) value and the power of exclusion (PE) value of the TPH gene-T457C locus were 0.624 and 0.187, respectively. The allele frequency showed significant difference from that of French people (P=0.04).@*CONCLUSION@#Polymorphism of the TPH gene-T457C locus could show ethnic and regional differences. It has a potential to be used in forensic science.

Influence of different types of posts and cores on color of IPS-Empress 2 crown / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To evaluate the influence of different types of posts and cores on the final color of the IPS-Emperss 2 crown.</p><p><b>METHODS</b>Five types of posts and cores (Cerapost with Empress cosmo, Cerapost with composite resin, gilded Ni-Cr alloy, gold alloy and Ni-Cr alloy) were made. The shifts in color of three points of IPS-Empress 2 crown surface (cervical, middle and incisal) with different posts and cores was measured with a spectroradiometer (PR-650).</p><p><b>RESULTS</b>The L* a* b* values of zirconium oxide and gilded Ni-Cr alloy posts and cores with ceramic crown were the highest. The L* a* values of zirconium oxide posts composite cores were higher while the b* values were lower. The L* a* b* values of Ni-Cr alloy were lower than that of gold alloy and were the lowest.</p><p><b>CONCLUSION</b>In combination with IPS-Empress 2 crown, zirconium oxide posts are suitable for routine use in the anterior dentition, and gilded Ni-Cr alloy and gold alloy posts and cores can be recommended for clinical practice. Ni-Cr alloy posts and cores can not be recommended for clinical practice.</p>

Cohort mortality study of dust exposed miners in iron mine / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the main diseases jeopardizing the health of the iron miners and to explore the relationship between dust exposure and malignancies as well as other diseases.</p><p><b>METHODS</b>A retrospective study with a cohort of 7,469 workers employed between January 1, 1972 and December 31, 1974 in Daye Iron Ore Mine Co. in Hubei Province was conducted. Standardized mortality ratio (SMR) was calculated for the main causes of death using Chinese national mortality rates for reference.</p><p><b>RESULTS</b>All subjects were followed up through December 31, 2003 with an accumulation of 199, 108.0 person years. A total of 1,752 workers died. The cumulative mortality was 23.5%. The cancers, cerebrovascular diseases, non-malignant respiratory diseases and cardiovascular diseases were main diseases that threatened workers' life span. The SMR for all subjects was a little higher than expected based on the Chinese national mortality rates. The diseases causing the significantly higher death rate were the nasopharynx cancer, liver cancer, lung cancer, pneumoconiosis and accident with SMR 1.84, 1.51, 1.83, 14.94 and 1.25 respectively. Increased mortality was observed among dust-exposed workers in the cohort. The cumulative mortality from all causes such as stomach cancer, lung cancer, nonmalignant respiratory diseases, cardiovascular diseases and accident in dust exposed workers were significantly increased compared with those in non-exposure workers with RR 1.35, 1.83, 1.61, 2.27, 1.34 and 1.69 respectively.</p><p><b>CONCLUSION</b>The risk factors especially dust exposure affect the health and lifespan of the iron mine workers.</p>

Influence of adhesion on the color of glass infiltrated alumina ceramic restorations / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the effects of luting agent on the final color of glass infiltrated alumina ceramic restorations.</p><p><b>METHODS</b>12 plate-shaped specimens with 12.5 mm in diameter and 0.5 mm thickness were fabricated from GI-II (color IG2). Vitadur alpha veneering porcelain (color A2) with 1.0 mm thickness was fired to GI- II glass/alumina composite. 12 plate-shaped background specimens simulating the metal alloy post-and-core 12.5 mm in diameter and 2 mm thickness were also made from Ni-Cr alloy. All-ceramic specimens were luted to the metal alloy by Zinc Phosphate cement, glass ionomer cement and composite resin. The color shifts of the specimens were measured by colorimeter.</p><p><b>RESULTS</b>Luting agents had effect on the final color of restorations. The influence of composite resin was least, followed by glass ionomer cement and Zinc Phosphate cement. The color difference between with and without Zinc Phosphate cement could be identified by the eye.</p><p><b>CONCLUSION</b>To reduce the effect of luting agents, composite resin is recommended to all-ceramic restorations' adhesion.</p>