Analysis of occupational health examination results of radiation workers in medical institutions of Shenzhen in 2020 / 中国辐射卫生

Objective To analyze the medical examinations of radiation worker in medical institutions and provide some basic data for radiation protection management. Methods The occupational health examination of 3568 radiation workers from 681 medical institutions who came to our hospital for occupational health examination from January 1 to December 31 in 2020 were summarized and analyzed. Results There was no case of suspected occupational radiation sickness. The abnormal rate was in the range of 2.17%～2.99%, the rate of occupational contraindicated was about 1.44%～2.17%. The total review rate was about 13.00%, more than 79.48% of the radiation workers were checked out other diseases or abnormal. The abnormal examination items are mainly ophthalmology, B ultrasound of liver, gallbladder, spleen and pancreas, liver function, electrocardiogram, blood routine, urine routine, blood pressure, B ultrasound of both kidneys and kidney function. The abnormal rate of ophthalmology in each level of institutions was decreased with the increase of the length of service, while the abnormal results of B-ultrasound of liver, gallbladder, spleen and pancreas, blood pressure, B-ultrasound of both kidneys and renal function were increased with the increase of service. Conclusion Maybe the radiation protection of radiation workers in medical institutions was well in Shenzhen, but there were different effects of the health status of the staff. Therefore, it is important to further strengthen the occupational health monitoring management.

Probiotics modulate the microbiota-gut-brain axis and improve memory deficits in aged SAMP8 mice

ProBiotic-4 is a probiotic preparation composed of , , , and . This study aims to investigate the effects of ProBiotic-4 on the microbiota-gut-brain axis and cognitive deficits, and to explore the underlying molecular mechanism using senescence-accelerated mouse prone 8 (SAMP8) mice. ProBiotic-4 was orally administered to 9-month-old SAMP8 mice for 12 weeks. We observed that ProBiotic-4 significantly improved the memory deficits, cerebral neuronal and synaptic injuries, glial activation, and microbiota composition in the feces and brains of aged SAMP8 mice. ProBiotic-4 substantially attenuated aging-related disruption of the intestinal barrier and blood-brain barrier, decreased interleukin-6 and tumor necrosis factor- at both mRNA and protein levels, reduced plasma and cerebral lipopolysaccharide (LPS) concentration, toll-like receptor 4 (TLR4) expression, and nuclear factor-B (NF-B) nuclear translocation in the brain. In addition, not only did ProBiotic-4 significantly decreased the levels of -H2AX, 8-hydroxydesoxyguanosine, and retinoic-acid-inducible gene-I (RIG-I), it also abrogated RIG-I multimerization in the brain. These findings suggest that targeting gut microbiota with probiotics may have a therapeutic potential for the deficits of the microbiota-gut-brain axis and cognitive function in aging, and that its mechanism is associated with inhibition of both TLR4-and RIG-I-mediated NF-B signaling pathway and inflammatory responses.

Identification, sequence analysis, and infectivity of H9N2 avian influenza viruses isolated from geese

The subtype H9N2 avian influenza virus greatly threatens the Chinese poultry industry, even with annual vaccination. Waterfowl can be asymptomatically infected with the H9N2 virus. In this study, three H9N2 virus strains, designated A/Goose/Jiangsu/YZ527/2011 (H9N2, Gs/JS/YZ527/11), A/Goose/Jiangsu/SQ119/2012 (H9N2, Gs/JS/SQ119/12), and A/Goose/Jiangsu/JD564/2012 (H9N2, Gs/JS/JD564/12), were isolated from domestic geese. Molecular characterization of the three isolates showed that the Gs/JS/YZ527/11 virus is a double-reassortant virus, combining genes of A/Quail/Hong Kong/G1/97 (H9N2, G1/97)-like and A/Chicken/Shanghai/F/98 (H9N2, F/98)-like; the Gs/JS/SQ119/12 virus is a triple-reassortant virus combining genes of G1/97-like, F/98-like, and A/Duck/Shantou/163/2004 (H9N2, ST/163/04)-like. The sequences of Gs/JS/JD564/12 share high homology with those of the F/98 virus, except for the neuraminidase gene, whereas the internal genes of Gs/JS/YZ527/11 and Gs/JS/SQ119/12 are closely related to those of the H7N9 viruses. An infectivity analysis of the three isolates showed that Gs/JS/SQ119/12 and Gs/JS/YZ527/11 replicated well, with seroconversion, in geese and chickens, the Gs/JS/JD564/12 did not infect well in geese or chickens, and the F/98 virus only infected chickens, with seroconversion. Emergence of these new reassortant H9N2 avian influenza viruses indicates that these viruses can infect both chicken and goose and can produce different types of lesions in each species.

Surface display of lead-binding protein PbrR on Escherichia coli and preliminary study of intestinal ;colonization by the recombinant bacteria / 中华微生物学和免疫学杂志

Objective To construct a recombinant Escherichia coli ( E. coli) with surface-dis-played lead specific binding protein PbrR and to further study intestinal colonization by the recombinant bac-teria in mice and gastrointestinal tolerance of the bacterial surface-displayed PbrR. Methods Chimeric pro-tein Lpp-OmpA coding sequence was chemically synthesized and inserted into the expression vector pET-21a to construct the outer membrane display vector pLOA. PbrR coding sequence was also obtained by chemical-ly synthesis and inserted into pLOA to generate the outer membrane display plasmid pLOA-pbrr. E. coli BL21 (DE3)pLysS was transformed with pLOA-pbrr and induced by IPTG. The expressed recombinant proteins were analyzed by 15% SDS-PAGE and Western blot assay. Lead adsorption capacity of the cell surface-dis-played PbrR in the simulated intestinal juice and tolerance of the recombinant E. coli to simulated gastric juice were analyzed, respectively. KM mice were orally given the induced recombinant bacteria by gastric lavage for 7 consecutive days and then were continually fed until day 30. The contents of recombinant bacte-ria in stool samples were detected by dilution plate method on day 7, 15 and 30. The recombinant protein with His tag was detected by immunoblotting on day 7 and 15. Results Based on Lpp-OmpA, the PbrR outer membrane display vector was successfully constructed. The recombinant fusion protein Lpp-OmpA-PbrR-His tag was highly expressed in E. coli. The recombinant E. coli strains displaying PbrR on their outer membrane accumulated a significant level of Pb2+ in simulated intestinal juice. Moreover, those strains showed a tolerance to gastric acid in vitro and could colonize in the intestinal tracts of mice via oral infection. The surface-displayed recombinant fusion protein showed a better tolerance to the environment of digestive tract. Conclusion The recombinant E. coli strain displaying PbrR on its surface showed a stronger capabili-ty of lead accumulation from simulated intestinal environment and could colonize in the intestinal tracts of mice. The surface-displayed recombinant PbrR also showed a good tolerance to digestive juice. This study paved the way for further researches on the selective elimination of lead by biosorption based on animal mod-els.

Study on microtiter plate kinetic method to detection ALT / 国际检验医学杂志

Objective To investigate the clinical application of ALT detection based on microtiter plate kinetic method in the au‐tomatic enzyme immunoassay system .Methods By beference to the IFCC recommended kinetic method ,microtiter plate kinetic method was established in the automatic enzyme immunoassay system of ALT detection .The linear ,intro‐batch and inter‐batch du‐plicability of the method were evaluated .ALT test results of 823 samples with microtiter plate kinetic method and automatic bio‐chemistry analyzer method were compared .Results Microtiter plate kinetic method had good linearity where ALT activity <303 U/L ,the intra batch and inter batch variation coefficients < 1/5TEa .The detection results of the clinical samples were correlated with the biochemical analyzer .Conclusion Microtiter plate kinetic method to detection ALT in the automatic enzyme immunoassay system is an ideal method for simultaneous detection of ALT and ELISA in large batch samples ,which is worth popularizing .

Effects of indium on micronucleus formation in human peripheral blood lymphocytes / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the cytotoxicity of indium chloride (InCl₃) and its effects on micro-nucleus formation in primary human lymphocytes cultured in vitro.</p><p><b>METHODS</b>The CCK-8 assay was used to evaluate the cytotoxicity of 24 h exposure to different concentrations of InCl₃(4, 40, 80, 200, 500, and 1 000 µmol/L) in lymphocytes cultured in vitro. The cytokinesis-block method was used to determine the micronucleus level in lymphocytes exposed to different concentrations of InCl₃and the effects of anti-oxidant vitamin C on micronucleus frequency.</p><p><b>RESULTS</b>Lymphocytes exposed to InCl₃of no less than 500 µmol/L had significantly lower survival rates than those in the control group (P < 0.05). Lymphocytes exposed to 80 µmol/L InCl₃had a significantly higher micronucleus frequency than those in the control group (P < 0.05). However, there was no further increase in micronucleus frequency of lymphocytes exposed to 200 µmol/L InCl₃. Lymphocytes cultured in whole blood and exposed to 500 or 1000 µmol/L InCl₃had a significantly increased micronucleus frequency than those in the control group (P < 0.001). The increase in micronucleus frequency of lymphocytes induced by indium could be partially antagonized by 20 or 100 µmol/L vitamin C.</p><p><b>CONCLUSION</b>InCl₃can induce an increase in micronucleus frequency of primary human lymphocytes cultured in vitro, which might be associated with DNA damage induced by oxidative stress.</p>

The significance of monitoring procalcitonin when applying antibiotics to trichlorethylene dermatitis / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the significance of monitoring procalcitonin (PCT) when applying antibiotics to trichlorethylene (TCE)-induced dermatitis.</p><p><b>METHODS</b>One hundred and two patients who were hospitalized and recovered from TCE-induced dermatitis in our hospital from 2006 to 2013 were enrolled as subjects. Based on whether the PCT level was monitored or not, we divided patients into regular group and PCT group. For the regular group, we applied antibiotic treatment and determined the course of treatment based on clinical symptoms, laboratory test results, medical imaging results, and bacterial culture. For the PCT group, in addition to the above treatments, antibiotic treatment was applied when the PCT level was not lower than 0.25 ng/ml and stopped when the PCT level was lower than 0.25 ng/ml. The distribution of bacterial infection sites, type of bacteria, type of antibiotics, average period of hospitalization, and course of antibiotic treatment were compared between the two groups.</p><p><b>RESULTS</b>There were no significant differences in the distribution of bacterial infection sites, type of bacteria, type of antibiotics, and average period of hospitalization between the two groups (P > 0.05). The course of antibiotic treatment for the PCT group was significantly shorter than that for the regular group (25.37 ± 11.66 vs 20.58 ± 7.53 d, P < 0.05).</p><p><b>CONCLUSION</b>Under similar conditions of bacterial infection, antibiotic treatment of TCE-induced dermatitis based on the serum PCT level can significantly shorten the course of treatment and avoid the abuse of antibiotics.</p>

Clinical factors and prognostic significance of skeletal related events in bone metastasis of non small cell lung cancer in 223 cases / 重庆医学

Objective To explore the risk factors of skeletal related events (SREs) in non small cell lung cancer with bone metastases and its effect on the prognosis .Methods Totally 223 cases of NSCLC patients with bone metastasis were retrospective studied from January 2010 to December 2012 in our hospital .The clinical features ,predictive factors for SREs were analysed by sin‐gle factor and multifactor analysis .Results Among 223 cases of NSCLC patients with bone metastasis ,119 cases occured with SREs(53 .4% ) .Univariate analysis showed that the occurrence of SREs in female ,no smoker ,adenocarcinoma ,solitary bone metas‐tasis lesions were less than the male ,smoker non‐adenocarcinoma ,and multiple bone metastases (P0 .05) .The multivariate analysis revealed only multiple bone metastases was an independent risk factor for SREs .The median survival time of the NSCLC patients with bone metastasis was 15 .3 months .Moreover ,survival analysis showed that SREs had no statistical significance on the prognosis of bone metastasis in NSCLC patients (P>0 .05) .Conclusion The female ,adenocarcinoma ,smoking history ,solitary bone metastasis lesions occurred in patients with lower risk SREs .Multiple bone metastasis is an independent risk factor for SREs ,attention should be paid to monitoring and prevention .

Myths and countermeasures for oncologic professional degree postgraduate cultivation / 中华医学教育探索杂志

Two kinds of incorrect cultivation modes were existed since the launch of cultivation for professional degree postgraduate of clinical medicine:taking professional degree postgraduates as scientific degree postgraduates to cultivate or taking scientific degree postgraduates as professional degree postgraduates to cultivate.In this paper,taking the cultivation of oncologic professional degree postgraduates as example,we put forward some suggestions:strengthening clinical skill training,promoting thinking mode of evidence-based medicine and enhancing clinical trial and statistical training.

Isolation and characterization of promoter of ADS from Artemisia annua / 中国中药杂志

<p><b>OBJECTIVE</b>To try to find the ways to enhance the expression of ADS gene encoding amorpha-4,11-diene synthase, a key enzyme in artemisinin biosynthesis pathway catalyzing the formation of amorpha-4,11-diene from farnesyl diphosphate, and accelerate the artemisinin synthesis, the promoter of ADS was isolated and characterized.</p><p><b>METHOD</b>5' untranslated regions of ADS were isolated from Artemisia annua with PCR. For functional characterization, the isolated fragment was fused with GUS reporter gene and introduced into Nicotiana tabacum by Agrobacterium-mediated transformation. The GUS expression regulated by 5' untranslated regions of ADS in transgenic N. tabacum under the normal or stressed conditions were detected by histochemical staining and quantitative spectrophotometry assay.</p><p><b>RESULT</b>The 2 448 bp DNA fragment upstream of ADS coding sequence was isolated from A. annua and introduced into N. tabacum. Histochemical staining showed that the isolated fragment conferred stable GUS expression in transgenic plants. The quantitative results showed that the GUS activity in transgenic tobacco plants treated by low-temperature (4 degrees C) and ultraviolet irradiation were 1. 6 and 2.2 folds higher than that in the controls.</p><p><b>CONCLUSION</b>It was suggested that the isolated fragment had promoter activity and maybe responsive to adverse environmental stresses.</p>

Expression of peroxisome proliferator-activated receptors in the epidermis of skin from patients with psoriasis vulgaris / 中华皮肤科杂志

Objective To investigate the expression of peroxisome proliferator-activated receptors (PPARs) in epidermal keratinocytes of patients with psoriasis vulgaris and its significance.Methods Immunohistochemical method was used to examine the expression of PPARα,β/δand.γ in tissue specimens from the normal skin of 5 human controls,lesional and normal skin adjacent to the lesions of 17 patients with psoriasis vulgaris.A semi-quantitative analysis was carried out by image analyzer.Results Different levels of expression of the three PPAR isotypes were observed in the nuclei of epidermal keratinocytes of normal human controls.The expression intensity of PPAR α,β/δand γ was statistically higher in the epidermis of adjacent normal skin than that in psoriatic lesions(all P<0.01)and normal control skin(all P<0.01).Conclusions The decreased expression of PPARα may be associated with the overproliferation and parakeratosis of epidermal keratinocytes in psoriatic lesions,and PPAR β/δ and γ may display a synergistic effect on the maintenance of homeostatic proliferation and difierentiafion of epidermal cells.

Screening and optimizing the colorectal cancer related tumor markers from multi-tumor markers proteinchip / 肿瘤研究与临床

Objective To analyze the association between C12 tumor markers and colorectal cancer,in order to screen for colorectal cancer related tumor markers so as to provide theoretical base for the establishment of colorectal cancer diagnostic biochips. Methods The sera of 173 colorectal cancer patients were detected for 12 common tumor markers including carcinoembryonic antigen (CEA), alpha-fetoprotein (AFP), carbohydrate antigen 19-9 (CA19-9), carbohydrate antigen 242 (CA242), cancer antigen 15-3 (CA15-3), cancer antigen 125 (CA125), prostate specific antigen (PSA), free-PSA, neuron-specific enolase (NSE),human chorionic gonagotropin-beta (β-HCG), human growth hormone (HGH), and ferritin using the C12tumor markers proteinchip, and colorectal cancer related parameters were analyzed by Kappa test and cost-effectiveness analysis to find the most optimal tumor marker program. Results CEA (36.4 %), CA242(19.7 %), CA19-9 (18.5 %), CA125(9.8 %) were major tumor markers increased among the 173 colorectal cancer patients. Kappa test revealed 7 tumor marker programs having strong consistency with the detection results of C12 tumor markers proteinchip, and CEA singly detected was proved to be the best program by cost-effectiveness analysis. Conclusion C12 tumor markers proteinchip system have limited value in the diagnosis of colorectal cancer, but the design of chip is too complicated and costly for widespread screening among the high risk populations. Searching for new colorectal cancer biomarkers and designing small diagnostic chip could significantly enhance the clinical value of tumor markers in terms of diagnostic rate and practical utility.

Construction and application of a new prokaryotic expression vector derivative of pBV220 / 生物工程学报

A single-stranded oligonucleotides containing a 6 histidine sequence, a hydroxylamine cleavage site, a thrombin cleavage site, and stop codon TAA were inserted into the polylinker's downstream of prokaryotic expression vector pBV220 between BamHI and PstI. The resultant vector is named pBV223. Proteins expressed in this vector will have a 6 histidine tail as affinity handy fused to their C terminus and can be quickly purified by one step immobilized metal affinity chromatography (IMAC). This plasmid is verified by restriction map and DNA sequencing. Subsequently, the metastasis suppressor gene nm23-H1 cDNA (without the stop codon) was cloned into vector PBV223 in frame with the 6-histidine sequence, hydroxylamine and thrombin cleavage sites. The soluble nm23-H1 fusion protein was successfully induced in the bacterial DH5a and easily purified with Ni chromatograph. These results indicated that the strategy to clone the single-stranded oligonucleotides directly into the restriction sties between BamH I and Pst I in the pBV220 vector is the simplest and cost-effective method.

Prokaryotic expression, purification and identification of human mutant nm23-H1 protein / 重庆医科大学学报

Objective:To construct prokaryotic expression vector of mutant nm23-H1(S120G,S120G-P96S, S44A), and then express and purify the proteins. Methods: Mutant nm23-H1 fragments were amplified by PCR. The prokaryotic expression vectors of pET28a-nm23-H1 were constructed by gene recombination technique and verified by restriction enzyme analysis and sequencing. The positive clones were transformed into E. coli BL21(DE3) and soluble analysis of the expression was conducted in these systems. The proteins were purified by nickel column chromatography and identified by Western blot. Results: The sequences and open read frames of all the pET28a-nm23-H1 were completely correct. After transforming, these plasmids could express the target proteins. The protein production was very high. Among these mutant proteins, S44A was soluble expression and S120G was partly soluble. However, S120G-P96S was mostly in the inclusion bodies. The molecular weight of these mutant proteins was 20 kD detected by Western blot, which was as the same as the objective protein. Conclusion: We have succeeded in constructing the prokaryotic expression vectors of pET28a-nm23-H1(S120G, S120G-P96S, S44A), and the S120G and S44A proteins can be used in following studies. However, S120G-P96S protein which expressed in this system may not be suitable for the following studies.

Construction, expression and purification of kinase suppressor of Ras, KSR / 中国肺癌杂志

<p><b>BACKGROUND</b>Ras to MAPK pathway plays a critical role in the transmission of many growth and developmental signals. A new component of this pathway which is termed kinase suppressor of Ras (KSR) was found in 1995. KSR is as a scaffolding protein that coordinates the assembly of a multiprotein complex containing mitogen-activated protein kinase (MAPK) and its upstream regulators. It has been proven that KSR has many phosphorylation sites, and phosphorylation state changes response to signaling events. Site-directed mutagenesis can precisely change the base sequence and get mutant proteins. The aim of this study is to construct two mutant proteins of KSR by using site-directed mutagenesis, and to express and purify them, therefore to provide basement for studying the functional and biochemical mechanisms of KSR.</p><p><b>METHODS</b>Site-directed mutagenesis of pCMV-Tag2b-KSR gene was performed by modified QuikChangtm site-directed mutagenesis kit method. Two pairs of mutagenic primers were synthesized in vitro and two mutations desired, the recombinant plasmids were verified by restriction enzyme analysis and DNA sequencing. Then positive clones were transfected into 293T cell line. The purified mutant proteins were analyzed by Western blot.</p><p><b>RESULTS</b>Two mutants were successfully constructed. The results of DNA sequencing confirmed that the base sequences of the mutant genes were completely concordant with experiment design, which could be used to be transfected into 293T cell line. The purified mutants were identified by Western blot.</p><p><b>CONCLUSIONS</b>Two mutant KSR genes are successfully constructed. It provides experimental basement for further functional research of KSR.</p>

Clinical significance of multiple tumor marker protein chip in monitoring the recurrence,progression and metastasis of lung cancer / 中国肺癌杂志

<p><b>BACKGROUND</b>According to the report of the 11th World Conference on Lung Cancer, lung cancer is the leading cause of cancer related death. So there is important clinical significance to monitor the patients with lung cancer through different ways. The aim of this study is to investigate the clinical significance of multiple tumor marker protein chip in monitoring the recurrence, progression and metastasis of lung cancer.</p><p><b>METHODS</b>Forty-four patients were selected, who were detected at least 4 times with tumor mar-ker protein chip. Based on efficacy and status, patients were classified as six grades. Correlation of expression level of each tumor marker with grade of efficacy and status was analyzed. And the discriminant functions for recurrence, progression and metastasis of lung cancer were established.</p><p><b>RESULTS</b>Grade of efficacy and status was closely related to CA199, CEA, CA242, AFP and CA125 in adenocarcinoma (AC), to CA125 in squamous cell carcinoma (SqCa), and to CA199 and CA125 in small cell lung cancer (SCLC). Based on the discriminant functions, accuracy rate of efficacy and status judgement was 89.4%, 80.4%, 78.3% and 66.7% for female AC, male AC, SqCa and SCLC respectively.</p><p><b>CONCLUSIONS</b>There is important clinical significance of multiple tumor marker protein chip in monitoring the recurrence, progression and metastasis of lung cancer (especially adenocarcinoma).</p>

Effects of nm23-H1 point mutation on activity of GSK-3β in human high-metastatic large cell lung cancer cell line L9981 / 中国肺癌杂志

<p><b>BACKGROUND</b>The results of our previous studies have proven that nm23-H1 gene can suppress metastasis of lung cancer, which may be associated with suppression of the Wnt signal pathway through up-regulating the activity of glycogen synthase kinase 3β (GSK-3β), the key kinase of the Wnt signal pathway. The aim of this study is to investigate the effect of point mutation of nm23-H1 gene on GSK-3β activity in cytoplasm and nucleus in human high-metastatic large cell lung cancer cell line L9981.</p><p><b>METHODS</b>Using immunoprecipitation and a radioactive isotope scintillation counter, the activity of GSK-3β was detected in cytoplasm and nucleus of human low-metastatic large cell lung cancer cell line NL9980, human high-metastatic large cell lung cancer cell line L9981, L9981-pEGFP (transfected with vector), L9981-nm23-H1-pEGFP (transfected with wild type nm23-H1), L9981-nm23-H1 S44A -pEGFP mutant (transfected with serine 44 to alanineon of nm23-H1 gene), L9981-nm23-H1 P96S -pEGFP mutant (transfected with proline 96 to serine of nm23-H1 gene), L9981-nm23-H1 H118F -pEGFP mutant (transfected with histidine 118 to phenylalanine of nm23-H1 gene) and L9981-nm23-H1 S120G -pEGFP mutant (transfected with serine 120 to glycine of nm23-H1 gene).</p><p><b>RESULTS</b>The GSK-3β activity in cytoplasm and nucleus was remarkably decreased in the transgene lung cancer cell lines transfected with mutant nm23-H1 cDNA (L9981-nm23-H1 S44A -pEGFP, L9981-nm23-H1 P96S -pEGFP, L9981-nm23-H1 H118F -pEGFP and L9981-nm23-H1 S120G -pEGFP). Significant differences of GSK-3β activity in cytoplasm and nucleus were observed (P < 0.05) when L9981-nm23-H1-pEGFP cell line was compared with L9981-nm23-H1 S44A -pEGFP, L9981-nm23-H1 P96S -pEGFP, L9981-nm23-H1 H118F -pEGFP and L9981-nm23-H1 S120G -pEGFP lung cancer cell lines. There was a highly significant difference in GSK-3β activity in the cytoplasm between L9981-nm23-H1-pEGFP cell line and L9981-nm23-H1 P96S -pEGFP lung cancer cell line (P < 0.01 ). A highly significant difference in GSK-3β activity in the nucleus was observed (P < 0.01) when L9981-nm23-H1-pEGFP lung cancer cell line was compared with L9981-nm23-H1 S44A -pEGFP, L9981-nm23-H1 P96S -pEGFP and L9981-nm23-H1 H118F -pEGFP lung cancer cell lines.</p><p><b>CONCLUSIONS</b>(1) Point mutation of nm23-H1 gene can significantly influence the regulating effects on the GSK-3β activity in the cytoplasm and nucleus in the human high-metastatic large cell lung cancer cell line L9981. (2) The effects of nm23-H1 gene on metastatic phenotype may be related to the upregulation of GSK-3β activity in human high-metastatic large cell lung cancer cell line L9981.</p>

Relationship between LAPTM4B gene polymorphism and susceptibility of lung cancer / 中国肺癌杂志

<p><b>BACKGROUND</b>It has been proved that lysosome-associated protein transmembrane 4 beta (LAPTM4B) plays an important role in tumorigenesis, the *2/*2 genotype of LAPTM4B was closely related to liver cancer susceptibity. The aim of this study is to investigate the possible association between the allelic variation of LAPTM4B and the genetic susceptibility of lung cancer.</p><p><b>METHODS</b>The genotype of LAPTM4B was detected in 131 patients with lung cancer and 104 unrelated healthy adult individuals as control by polymerase chain reaction based on special primers. The genotypic distribution of LAPTM4B was analyzed by Chi-square test.</p><p><b>RESULTS</b>The allelic frequencies of the *1 and *2 were 74.8% and 25.2% in the lung cancer group and 72.1% and 27.9% in the healthy control group respectively, but no significant difference was observed between the two groups (Chi-square=0.433, P=0.510). The genotypic distribution of the *1/*1,*1/*2 and *2/*2 were 53.4%, 42.7% and 3.8% in the lung cancer group, 54.8%, 34.6% and 10.6% in the healthy control group respectively, there was no significant difference between the two groups (Chi-square=4.89, P=0.087). No remarkable association was observed between the genotypic distribution of LAPTM4B and the clinical information in the patients with lung cancer, including pathological type and TNM staging.</p><p><b>CONCLUSIONS</b>The results suggest that the allele of LAPTM4B is not closely associated with genetic susceptibility of lung cancer.</p>

Construction of eukaryotic expression vectors of carboxyl terminus and amino terminus of kinase suppressor of Ras (KSR) and their expression in 293T cell line / 中国肺癌杂志

<p><b>BACKGROUND</b>The present experimental data have showed that the function of kinase suppressor of Ras (KSR) is mainly as a scaffold protein that coordinates the assembly of a multiprotein complex containing MAPK and its upstream regulators. But whether KSR has kinase activity is still the point of argument until now. The aim of this study is to construct eukaryotic expression vectors of carboxyl terminus and amino terminus of KSR and to detect their expression in 293T cell line.</p><p><b>METHODS</b>N-KSR and C-KSR were amplified by PCR. The eukaryotic expression vectors of pCMV-Tag2b-N-KSR and pCMV-Tag2b-C-KSR were constructed by gene recombination technique and the recombinant plasmids were verified by restriction enzyme analysis and sequencing. Then positive clones were transfected into 293T cell line. Expression of target proteins was analyzed by Western blot.</p><p><b>RESULTS</b>The sequences and open read frames of the two vectors were both completely concordant with experiment design. The target proteins could be observed in transfected 293T cells by Western blot.</p><p><b>CONCLUSIONS</b>Eukaryotic expression vectors of pCMV-Tag2b-N-KSR and pCMV-Tag2b-C-KSR are successfully constructed, and they can be expressed in 293T cells. It provides an experimental base for further research work.</p>

Effects of polysaccharid on expression of angiogenic-related genes in human high-metastatic large cell lung cancer cell line L9981 / 中国肺癌杂志

<p><b>BACKGROUND</b>Lung cancer is one of the most malignant cancers which is hazarding the people's health and life in the world. At present, it is a highlight to exploit antitumor drug from plant at home and abroad. The aim of this study is to observe the effects of polysaccharid (PS-T) on expression of angiogenic-related gene mRNA in human high-metastatic large cell lung cancer cell line L9981, and to explore its possible molecular mechanism.</p><p><b>METHODS</b>L9981 in vitro was cultured, and the growth data were obtained by trypan blue staining. The mRNA transcript expression of β-catenin, E-cadherin, TIMP-1, CD44V6, MMP-2, endostatin, VEGF was detected in L9981 by RT-PCR before and after treating with PS-T. The ability of invasion of L9981 was determined by Boyden chamber method.</p><p><b>RESULTS</b>(1)PS-T had remarkably inhibitive effects on the growth of L9981 in vitro. The inhibitive rate of PS-T on L9981 was concentration-dependent. No significant difference of inhibitive rate was found among the PS-T (1g/L), cisplatin (3mg/L) and PS-T (0.05g/L) + cisplatin (1.5mg/L)(P > 0.05). (2)The mRNA expression level of β-catenin, E-cadherin, TIMP-1, endostatin and MMP-2 was upregulated, while that of VEGF and CD44V6 was downregulated. Out of them the mRNA expression level of TIMP-1 and endostatin was remarkably upregulated, the expression level of CD44V6 was significanyly downregulated. (3)The in vitro invasive abilities of L9981 was significantly decreased in the PS-T, DDP and PS-T+DDP groups compared with that in blank control group.</p><p><b>CONCLUSIONS</b>(1)PT-S could inhibit the growth of human high-metastatic large cell lung cancer cell line L9981 in vitro, the effect is dose-dependent. (2)PS-T can down- or up-regulate the mRNA transcript expression of some angiogenic-related gene mRNA. (3)PS-T has remarkably coordinating effects with cisplatin in the L9981 lung cancer cell line.</p>