[An investigation of mental health in migrant workers in an enterprise].

Objective: To investigate the mental health status in migrant workers in a labor-intensive enterprise and related influencing factors. Methods: Typical sampling was used to perform an investigation in 910 migrant workers in a large foreign-funded labor-intensive enterprise in Shenzhen, China. All the respondents gave informed consent and completed the questionnaire independently and anonymously. The self-reported mental health status was evaluated using the Beck Anxiety Inventory, Beck Depression Inventory, and General Health Questionnaire. Results: Of all the migrant workers in this enterprise, 7.2% had a positive self-reported anxiety symptom, 25.4% had a moderate or severe self-reported depression symptom, and 76.4% had a poor self-reported general health status. Age had significant influence on the self-reported depression symptom (χ2=21.968, P<0.05) ; age did not have significant influence on the self-reported anxiety and general health status (χ2=6.616、12.498, both P>0.05) . The knowledge of occupational hazards had significant influence on mental health status (χ2Depression=47.289, χ2General health=21.087, both P<0.05) . The feeling of work had significant influence on self-reported depression and general health status (χ2Depression=52.406, χ2General health=17.327, both P<0.05) . Attention to self mental health had significant influence on self-reported depression (χ2=17.714, P<0.05) , and whether the person wanted to learn the knowledge of mental health had significant influence on self-reported anxiety (χ2= 6.145, P<0.05) . Conclusion: The self-reported mental health status in migrant workers is poor and is associated with age, worry about exposure to occupational hazard factors, emphasis on mental health knowledge, and a focus on personal mental health. Therefore, targeted occupational health education and occupational mental health education should be strengthened.

[Natural history of colorectal cancer: a Meta-analysis on global prospective cohort studies].

Objective: To acknowledge the availability and rates of annual transition of outcomes during the progression and regression stages of colorectal cancer (CRC) and related diseases, by pooling global follow-up studies on the natural history of CRC. Methods: Till March, 2017, data was collected through systematic literature review over multiple databases, including PubMed, Embase, Cochrane and Chinese Biology Medicine (CBM) disc. Information regarding the characteristics, classification system of health states, related outcomes and incidence rates on CRC or high-risk adenoma for the surveillance cohorts of the studies, were extracted and summarized. Both Meta and sensitivity analyses were performed on those outcomes if they appeared in more than 3 studies, using the random effects model. Annual transition rate with 95%CI was used to estimate each of the outcomes, Quality of the studies was assessed, using the Newcastle-Ottawa Scale. Results: A total of 29 cohort studies were included, with the mean follow-up period as 5.7 years. All studies except one, focused on adenoma-carcinoma pathway and reported the outcome parameters of adenomas by different risk, and some reported the findings on different sizes (n=6) of adenomas. These cohorts were divided into three groups (normal status, with low-risk or high-risk adenoma) according to the status of baseline endoscopic pathologic findings. Their available outcome parameters, corresponding number of involved articles, aggregated sample size and pooled annual transition rates were presented. Six parameters were obtained in the normal cohorts, including those from normal to low-risk adenoma (16 articles, 58 235, 0.030: 0.024-0.037), to high-risk adenoma (17 articles, 62 089, 0.003: 0.002-0.004), to diminutive adenoma (<5 mm, 4 articles, 1 277, 0.021: 0.013-0.029), to small adenoma (6-9 mm, 4 articles, 1 277, 0.006: 0.001-0.010), to large adenoma (≥10 mm, 7 articles, 3 531, 0.002: 0.000-0.003) and to CRC (19 articles, 104 836, 0.000 3: 0.000 2-0.000 5). Three parameters were obtained in low-risk adenoma in cohorts with polypectomy findings, including recurrence (9 articles, 4 788, 0.109: 0.062-0.157) from low-risk adenoma after polypectomy to high-risk adenoma (10 articles, 5 736, 0.009: 0.004-0.013) and to CRC (12 articles, 11 347, 0.000 6: 0.000 4-0.000 8). Three parameters were obtained on high-risk adenoma from cohorts with polypectomy findings, including recurrence (12 articles, 7 030, 0.038: 0.028-0.048) from high-risk adenoma after polypectomy to low-risk adenoma (8 articles, 2 489, 0.133: 0.081-0.185) and CRC (14 articles, 14 899, 0.002: 0.001-0.003). Except for normal to low-risk adenomas, results from the sensitivity analysis for the other parameters showed stable. Of the included studies, two presented incidence rates of CRC in different clinical stages and the another two were focusing on the parameters related to serrated pathway. Conclusions: Globally, follow-up studies reported data on natural history of colorectal cancer is of paucity. Compared to the "adenoma-carcinoma" pathway, transition parameters of the serrated lesion pathway are more limited. This Meta-analysis provided convincing evidence for optimizing the strategies regarding follow-up program on the disease, using the baseline endoscopic findings from global CRC Screening Program. These results also offered strong data-related support for Chinese population- specific interventional model on colorectal cancer.

Identification of alternative splicing variants of the beta subunit of human Ca(2+)/calmodulin-dependent protein kinase II with different activities.

The beta subunit of human Ca(2+)/calmodulin-dependent protein kinase II (beta CaMKII) was identified by searching through an expressed sequence tag database and rapid amplification of cDNA 5'-ends and was assigned to chromosome 7. Reverse transcription-polymerase chain reaction and sequencing analysis identified at least five alternative splicing variants of beta CaMKII (beta, beta6, betae, beta'e, and beta7) in brain and two of them (beta6 and beta7) were first detected in any species. When expressed in HEK 293 cells, the Ca(2+)/calmodulin-dependent kinase activity of beta7, the shortest variant, was much lower than that of either beta (the longest one) or betae (the medium one), suggesting possible regulation of beta CaMKII activity by alternative splicing.

Carboxyl terminal of rhodopsin kinase is required for the phosphorylation of photo-activated rhodopsin.

Human rhodopsin kinase (RK) and a carboxyl terminus-truncated mutant RK lacking the last 59 amino acids (RKC) were expressed in human embryonic kidney 293 cells to investigate the role of the carboxyl terminus of RK in recognition and phosphorylation of rhodopsin. RKC, like the wild-type RK, was detected in both plasma membranes and cytosolic fractions. The C-terminal truncated rhodopsin kinase was unable to phosphorylate photo-activated rhodopsin, but possesses kinase activity similar to the wild-type RK in phosphorylation of small peptide substrate. It suggests that the truncation did not disturb the gross structures of RK catalytic domain. Our results also show that RKC failed to translocate to photo-activated rod out segments. Taken together, our study demonstrate the carboxyl terminus of RK is required for phosphorylation of photo-activated rhodopsin and strongly indicate that carboxyl-terminus of RK may be involved in interaction with photo-activated rhodopsin.

Functional expression of opioid receptor-like receptor and its endogenous specific agonist nociceptin/orphanin FQ during mouse embryogenesis.

Expression of opioid receptor-like receptor (ORL1) and its endogenous peptide agonist nociceptin/orphanin FQ (N/OFQ) during mouse embryogenesis have been investigated. Transcripts of ORL1 and N/OFQ were detected by RT-PCR in mouse brain of day 8 embryo (E8) and the expression continued afterwards. Northern blot analysis revealed abundant expression of ORL1 at postnatal day 1 (P1) and N/OFQ at E17 and P1 in the brain but none was detected in other embryonic tissues. The presence of functional ORL1 in mouse embryonic brain was also confirmed by specific binding of [3H] N/OFQ (kd = 1.3 +/- 0.5 nM and Bmax = 72 +/- 9 fmol/mg protein) as well as by N/OFQ-stimulated G protein activation.

Significant down-regulation of alpha-albumin in human hepatoma and its implication.

The potential association of alpha-albumin (ALF) with hepatocellular carcinoma (HCC) was investigated. Expression of ALF was significantly reduced in HCC tumor tissue as compared with the paired peritumor tissue from 16 patients and in four HCC cell lines as compared with normal hepatocytes. ALF mRNA was also down-expressed in circulating HCC cells compared to circulating normal hepatocytes. The proliferation of Hep3B cells was inhibited by over-expression of ALF. Taken together, ALF is significantly down-regulated in HCC, and this might facilitate the proliferation of HCC. Thus, detection of ALF mRNA, in addition to that of alpha-fetoprotein (AFP) mRNA, might help to distinguish normal or malignant hepatocytes in peripheral blood.

Suppression of phospholipase C blocks Gi-mediated inhibition of adenylyl cyclase activity.

The potential effect of inhibition of phospholipase C on the response of Gi-coupled receptors was investigated in neuroblastoma x glioma hybrid (NG108-15) cells. The phospholipase C specific inhibitor 1-[6-((17beta-3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl]-1H -pyrrole-2,5-dione (U73122), which did not affect basal and forskolin-stimulated adenylyl cyclase activities, time- and dose-dependently blocked delta-opioid receptor-mediated inhibition of adenylyl cyclase activity, the EC50 (0.5 microM) of which was consistent with that for inhibition of bradykinin-dependent phospholipase C activation (EC50 = 1 microM). U73122 treatment also blocked functional responses of m4 muscarinic receptor and alpha2-adrenoceptor in NG108-15 cells and three opioid receptors (mu, delta and opioid receptor-like receptor (ORL1)) in human neuroblastoma SK-N-SH cells. 1-[6-((17Beta-3-Methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl]-2, 5-pyrrolidinedione (U73343), an inactive analog of U73122, did not show any effect, which suggests that the blockade by U73122 of Gi-coupled receptor-mediated signaling is probably mediated through inhibition of phospholipase C, although a possible direct modification of G proteins can not be excluded. Furthermore, treatment with U73122 but not U73343 blocked the GTP-induced inhibition of adenylyl cyclase, indicating blockade at the level of Gi proteins.

Production, purification, and characterization of an intracellular aflatoxin-detoxifizyme from Armillariella tabescens (E-20).

Some Armillariella tabescens (E-20) multienzymes have previously been reported to present detoxifying activities against aflatoxins. In this paper, we describe the isolation purification of an intracellular enzyme, named aflatoxin-detoxifizyme, which exhibited detoxification activity on aflatoxin B(1) (AFB(1)). This aflatoxin-detoxifizyme exhibited a specific activity of 7.09 nmol min/mg at pH 6.0 and 28 degrees C. The apparent molecular mass was 51.8 kDa as determined by SDS-PAGE. The isoelectric point was estimated to be 5.4 and optimum activity for the enzyme was found at pH 6.8 and 35 degrees C. The activity of the purified enzyme was confirmed by Ames test.

Metabolism of 6-chloro n-butyl phthalide in rat.

6-Chloro n-butyl phthalide (CBP) was orally administered to healthy, male Wistar rats pretreated with or without 3-methylcholanthrene (3-MC) by a single dose of 150 mg/kg, and urine samples were collected for 0-24 h. The urine sample was hydrolyzed with beta-glucuronidase, extracted and concentrated for TMS derivatization, and analysed on a GC-MS system for identification of CBP metabolities. Mass spectral analysis suggests that 7 CBP metabolites were present in the urine sample, and similar metabolism patterns were viewed in rats with or without pretreatment with 3-MC. Four main metabolites of CBP in rat urine were identified as alpha-beta oxolate, beta-gamma oxolate, beta-hydroxylate and gamma-hydroxylate, based on their chromatographic and mass spectral properties. Two hydroxylates have been previously identified in CBP metabolism by rat liver microsomes. The other two metabolites with higher polarity were tentatively identified as dihydroxylation products on the n-butyl side chain by the mass spectra of their TMS derivatives. One minor metabolite was found by the isotopic effect of chlorine, but its specific structure was undetermined. The difference between in vivo and in vitro metabolic profiles of CBP is also discussed.

[The sequence analysis of the 3' end of intron 15 from human LDL receptor gene and its application in the study of gene diagnosis for familial hypercholesterolaemia].

To understand intron 15 of human LDL receptor gene, the DNA fragments from exon 15 to exon 16 and the 3' end of intron 15 were amplified with long chain PCR and anchored PCR. The 3' end of intron 15 was sequenced with Dynalbeads-Streptavidin Solid Phase technique. The sequence analysis showed that the 3' end of intron 15 contained the 3' splicing site and the branch site at 31 nucleotides upstream of the 3' end. Besides the authentic branch site, it is possible that the 3' end of intron 15 contains a cryptic site (GCCTCAC) at 20 nucleotides upstream of the 3' end. The sequences suggest that the PvuII polymorphism at Intron 15 is caused by the T-C substitution. According to the sequences of the 3' end, the new PCR-RFLP protocol for detection of PvuII polymorphism at intron 15 was developed. Using this protocol a representative familial hypercholesterolaemia family was identified with linkage analysis of PvuII polymorphism at intron 15.

[The function of T7 promoter as cis-acting elements for polymerase II in eukaryotic cell].

Using the chlorampheniol acetyltransterase gene as reporter, the function of phage T7 promoter in mammalian cells was studied by inhibition of transcription with alpha-amanitin. The experiment proved that the reporter under T7 promoter was transcribed by RNA polymerase II. Competitive electropho retic mobility shift assay (CEMSA) with TATA box, CAAT box, GC box and octamer showed that the TATA box was competitive molecular for synthetic T7 promoter. It is possible that T7 promoter is bound with TF II D transcription factor. The TATA box and octamer were inserted into Pvu II site upstream from the T7 promoter of pT7CAT. Two recombinant plasmids, pT7TATACAT and pT7OCTCAT, were constructed and transfected into CHO cells. CAT-activity test showed that T7 promoter strength was increased by octamer factor, not by TATA box. These results suggested that T7 promoter functions as cis-acting elements of RNA polymerase II transcriptional system in eucaryotic cells.

[Determination of some androgens and anabolic steroids in human urine by HPLC].

A simple method for determining androgens and anabolic steroids in human urine by HPLC has been developed. The compounds studied are nandrolone, methandienone, testosterone, methyltestosterone, testosterone propionate and nandrolone phenylpropionate. The stationary phase used is C8 bonded silica. Isocratic elution was done with CH3OH-CH3CN-H2O (4:5:6) and programmed flow. Detection limit can be less than 1 ng at the wavelength of 254 nm. The standard curves for each steroid have been set using internal standard (progesterone) and peak height ratio. Linear relationship exists between the ratio and concentration for each steroid. High and stable recovery has been achieved using Sep-Pak C18 cartridges for urine sample clean-up. The operation of the method is simple. The enzymatic hydrolyzation of conjugated steroids in human urine has also been investigated.

[Investigation on in vivo metabolism of 6-methoxy n-butyl phthalide].

A GC-MS method for the investigation on the metabolism of 6-methoxy n-butyl phthalide (MBP) is described. After oral administration of MBP, the rat urine sample was collected, hydrolyzed with beta-glucuronidase, extracted and concentrated for TMS derivatization, and then analyzed by GC-MS. MBP and its six oxidative metabolites were determined in the 0-24 h rat urine sample. The mass spectra of the metabolites and their derivatives were presented and the in vivo metabolic pathway was discussed.

[Investigation on in vivo metabolism of n-butyl phthalide].

A GC-MS method for the investigation on the metabolism of n-butyl phthalide (NBP) is described. After oral administration of NBP to rats, urine was collected, hydrolyzed with beta-glucuronidase, extracted and concentrated for TMS derivatization, and then analyzed by GC-MS. HBP and its four oxidative metabolites were determined in 0-24 h, 24-48 h rat urine. The mass spectra of the metabolites and their derivatives were presented and the in vivo metabolic pathway was discussed.

[In vitro metabolic studies of the novel anti-anxietic drug AF-5 and its metabolites in human liver microsome incubation system].

AIM: To study the metabolism of a novel anti-anxietic drug AF-5 and its metabolites (I, II) in human liver microsome incubation system. METHODS: Human liver microsomes were prepared, the enzyme activity was determined to be 8.79 mg.mL-1 by Lowry's method. The human liver microsome incubation system consisted of: human liver microsomes 2 mg.mL-1, glucose-6-phosphate (G-6-P) 0.01 mmol.mL-1, glucose-6-phosphate dehydrogenase (G-6-PDH) 1 U.mL-1, magnesium chloride (MgCl2) 4.0 mumol.mL-1, coenzyme II in oxidized form (NADP) 0.5 mumol.mL-1, and coenzyme I in reduced form (NADH) 1.0 mumol.mL-1. Two milligrams of AF-5 solubilized by Tween 80 was then added, the mixture was diluted to 5 mL with Tris-HCl solution and the mixture was incubated in a 37 degrees C water bath with shaking. Oxygen was passed over the liquid surface for 0.5 min every 20 minutes. The incubation was carried out for 40 min and 100 min respectively. Three volumes of ethyl ether were added to stop the metabolism, and more ethyl ether was used to extract the metabolites for 3 times. The ether extracts were pooled together, dried with anhydrous sodium sulfate, then evaporated to dryness. The residue was dissolved in 0.5 mL n-hexane and analyzed by GC/MS under the following conditions: 150 degrees C (1 min)[formula: see text]180 degrees C (1 min)[formula: see text]260 degrees C (2 min), in the total ion current mode, EI: 70 eV, interface temperature: 250 degrees C, ion source temperature: 200 degrees C. RESULTS: Two major metabolites were found and identified in this incubation system, and demonstrated that the in vitro metabolic pathway was that the carbon 4 was first oxidized to hydroxyl group, then further oxidized to a carbonyl group. CONCLUSION: In human liver microsome incubation system AF-5 was completely metabolized in 100 min to the hydroxy derivative I and carbonyl derivative II, with hydroxymetabolite as the major metabolite. Metabolite I was further transformed to metabolite II, which was not metabolized any further by the human liver microsomes.

[GC-MS analysis of norethandrolone and its metabolites in man].

Steroids in human urine were adsorbed on a macroporous XAD-2 resin, eluted with methanol, hydrolyzed with glucuronidase, extracted and concentrated for TMS derivatization and then analyzed with GC-MS. Norethandrolone (N) and its 9 metabolites were detected in urine samples 9-35 h after oral administration. Met-2 and Met-7 could be detected even in 84 h urine sample. The structures of 7 metabolites were elucidated and the variations of their concentration in urine were determined. The specific metabolites and characteristic ions for screening N positive urine were chosen. This method is sensitive enough; the detection limit of N was 10 ng per ml urine, i.e. 10 ppb.

[Investigation on the metabolism of dehydrochloromethyltestosterone in man].

A GC-MS method for the investigation on the metabolism of dehydrochloromethyltestosterone (DHCMT) in man is described. Steroids in human urine were adsorbed on a macroporous XAD-2 resin, eluted with methanol, hydrolyzed with glucuronidase, extracted and concentrated for derivatization and then analyzed with GC-MS. DHCMT and its 7 metabolites were detected in urine samples 8 to 30 h after oral administration. Their structures and the variation of their concentration in urine were determined and the possible metabolic pathways of DHCMT were proposed on the basis of GC-MS feature of its metabolites. The specific metabolites and characteristic ions for screening DHCMT positive urine were chosen. Furthermore, the influence of the sample pretreatment methods on GC-MS results has been studied.

[Separation of epimers of anisodamine (654-2) with HPLC using chiral additives in the mobile phase].

The separation of epimers of anisodamine were studied with HPLC using a reversed phase ODS C18 column. Several chiral additives were tried in the mobile phases. d-Camphorsulfonic acid and L-(+)-dibutyl tartrate showed good resolution ability for the separation of the isomers, a pair of epimers of anisodamine were well separated; while beta-cyclodextrin appeared to have no selectivity for the epimers.

[Gas chromatographic/mass spectrometric analysis of boldenone urinary metabolites in man].

The metabolism of boldenone (17 beta-hydroxy-1,4-androstem-3-one) in man has been investigated by gas chromatography/mass spectrometry. After oral administration of a 20 mg dose to man, six metabolites were detected in the conjugated fraction of the urinary samples. Boldenone, the major compound excreted in urine, was detected within 34 h after administration. In addition, several metabolites, resulting from the hydroxylation of boldenone and the reduction of the unsaturated carbon bonds of boldenone, were detected in the urine samples varying from 9 to 83 h. Extraction and fractionation of these metabolites were achieved by using XAD-2 column and gas chromatography. The recovery of the whole procedure was studied. Furthermore, the mass spectra of the metabolites are presented and major fragment pathways are discussed.