A human myeloma cell line(IBTCHM8310) obtained from the bone marrow ofa patient who was diagnosed as multiplemyeloma has been repropagated to datefor 270 generations within 50 months.The cells recovered from cryopreserva-tion in liquid nitrogen for 17 times,grewand reproduced well.The growth curve shows that themultiplication of the cells were slow onthe first two days,then went straightly upand culminated on the 6th and 7th days.Chromosome analysis was performed on96 cells on metaphases of 164th generationand it was between 63-69.There was stilla prominent mode 65,66.Of each 1?10~5/mlcells suspension,0.5ml was injected in tra-peritoneally to nude mice,Significant tu-mor masses appeared 3 weeks after injec-tion.Pathological section showed that thecells were identical to those of their pa-rent line.

Carcinoma, Non-Small-Cell Lung ; genetics ; Exons ; genetics ; Genes, erbB-1 ; genetics ; Humans ; Lung Neoplasms ; pathology

OBJECTIVETo evaluate the effect of antisense monocyte chemotactic protein-1 (A-MCP-1) nanoparticles (NPs) as gene carrier on gene transfer in two kinds of animal models.

METHODSPoly (lactic acid-co-glycolic acid) (PLGA) was used to make the NPs loaded with A-MCP-1 through a double-emulsion/solvent evaporation technique. NPs size was assessed by dynamic laser defractometer. The particle morphology was observed by scanning electron microscopy. DNA content in the NPs was measured by dissolving known amounts of NPs in chloroform and extracting DNA with water. In vitro release was performed in tris-EDTA buffer at 37 degrees C using double-chamber diffusion cells. The receiver buffer was replaced daily. The A-MCP-1 NPs was transfected into the cultured smooth muscle cells. PCR was used to evaluate the transfection of A-MCP-1. Cationic lipid (Lipofectamine) was used to transfect A-MCP-1 as control. After 48 hours incubation, cells were digested and examined by polymerase chain reaction. Twenty New Zealand white rabbits under jugular vein to artery bypass grafting procedure were divided into four groups: the first group received grafts treated with A-MCP-1 NPs, the second group received grafts treated with cationic liposome (dioleoyl trimethyl ammonium propane)-A-MCP-1, the third group received grafts treated with plasmid DNA, and the fourth group received grafts without transfection as control. Fourteen days after surgery the grafts were harvested. The expression of A-MCP-1 and its effect on MCP-1 in vein grafts were detected by dot blotting. The morphology of the grafts was investigated. To establish abdominal aortic aneurysms rats model, rats were randomly divided into three groups: A-MCP-1 NPs injection group, shame NPs injection group and control groups (without injection). Two weeks after surgery, diameter of abdominal aorta was measured and aortic tissue was obtained for PCR analysis to evaluate the A-MCP-1 expression. Western blot were applied to detect the inhibitory effect to the expression of MCP-1 mRNA and CD68 protein by A-MCP-1 NPs.

RESULTSNPs size ranged 198nm to 205nm with average around 201.4 nm. DNA content in the NPs was 4.14%. NPs showed steady release rate in vitro in Tris-EDTA solution. It released faster in the first week then maintained a slowly sustained release up to 16 days. In cell culture A-MCP-1 gene successfully transfected into smooth muscle cells by NPs vector. In vein grafting animal model, A-MCP-1 expression was detected in the vascular walls of NPs and cationic lipid treated groups. The degree of vascular hyperplasia in the gene NPs treated group was significantly lower than that in control group. There was no significant difference in the inhibition of intimal hyperplasia between NPs and cationic lipid treated groups. Two weeks after transfection in abdominal aortic aneurysm rats models, the abdominal aortic diameter of A-MCP-1 NPs injection group was (1.79 +/- 0.12) mm, significantly smaller than that of control groups [shame NPs group was (2.58 +/- 0.21) mm, and saline group was (2.63 +/- 0.29) mm] (P < 0.01). The expressions of MCP-1 mRNA and CD68 protein in A-MCP-1 NPs injection group were 12.5 +/- 1.5 and 17.6 +/- 2.1, which were much lower than those in control group [in shame NPs group, which were 35.7 +/- 4.5, 42.3 +/- 5.7 (P < 0.01), and saline group which is 32.4 +/- 3.9, 39.8 +/- 4.8 (P < 0.01)]. Specific band of A-MCP-1 was detected only in the A-MCP-1 NPs injection group by PCR.

CONCLUSIONA-MCP-1 gene NPs can be successfully used in rabbit vein grafting model and abdominal aortic aneurysm rats models, and may be potentially applied in clinical practice.

Animals ; Aortic Aneurysm ; genetics ; Chemokine CCL2 ; genetics ; metabolism ; Gene Transfer Techniques ; Genetic Vectors ; Lactic Acid ; chemistry ; Models, Animal ; Nanoparticles ; Oligonucleotides, Antisense ; genetics ; Polyglycolic Acid ; chemistry ; Polymers ; chemistry ; Rabbits ; Rats ; Transfection

Objective - To analyze the prevalence and influencing factors of work related musculoskeletal disorders (WMSDs) Methods among painters in the manufacturing industry. A total of 639 painters from one shipbuilding enterprise, one automobile manufacturing enterprise and three wooden furniture manufacturing enterprises in Guangdong Province were selected as the research subjects using typical sampling method. The Chinese version of Musculoskeletal Disorders Questionnaire was Results used to investigate the prevalence of WMSDs in the past one year, and the influencing factors were analyzed. The total prevalence rate of WMSDs among painters in the manufacturing industry was 37.4%. The prevalence of WMSDs in different vs vs P industries from high to low was shipbuilding, automobile and furniture manufacturing (50.0% 38.7% 29.0%, <0.01). The prevalence of WMSDs in different parts of the body from high to low was neck, ankle/foot, shoulder, low back, upper back, knee, vs vs vs vs vs vs vs vs P hand/wrist, hip/leg and elbow (20.7% 19.2% 17.4% 15.8% 14.1% 13.8% 13.5% 9.5% 6.6%, <0.01). Multivariate logistic regression analysis results showed that working in uncomfortable postures was a risk factor for neck, ankle/ P P foot and shoulder WMSDs (all <0.01); long time head turning was a risk factor for neck and shoulder WMSDs (both <0.05); P overweight and obesity, and bending and turning frequently at the same time were risk factors for ankle/foot WMSDs (all <0.05); P adequate rest time was a protective factor for neck and ankle/foot WMSDs (both <0.01); participated in physical exercise more P than once a week was a protective factor of neck and shoulder WMSDs in painters (all <0.05), after excluding the influence of Conclusion confounding factors. The prevalence of WMSDs in manufacturing painters was high, and the main body parts E mail 4813545@qq.com E mail wangzhongxu2003@163.com· · 中国职业医学 年 月第 卷第 期 ， ， ， 482 2022 10 49 5 Chin Occup Med October 2022 Vol.49 No.5 involved were neck, ankle/foot and shoulder. The influencing factors include individual factors, poor ergonomics factors and unreasonable work organization.

To establish a fluorescent quantitative PCR method (FQ-PCR) with TaqMan probe for simultaneous detection of polyomavirus (BKV) and cytomegalovirus (CMV) and to evaluate its clinical application in the renal transplantation recipients. The conservative sequences of BKV and CMV were targeted and amplified by nested PCR technique. The PCR products were cloned into the plasmids pcDNA3. 1(+). The recombinant plasmid containing target sequences of BKV and CMV were constructed as external standards. The TaqMan-based assay was optimized. For evaluating the assay, the sensitivity was determinated by diluted standard (5 X 103-10icopies/mL), and the specificity was verified by negative control and positive control, and the precision was assessed by intra-assay coefficient of variation (ICV) through detecting standard repeatedly (20 times). A total of 480 blood samples of renal transplantation recipients were used to detect BKV and CMV DNA simultaneously with FQ-PCR, and the concentrations of FK506 were measured by ELISA. The association of DNA copy and concentrations of FK506 was analyzed. The cloned target BKV and CMV DNA was confirmed by sequencing and analysis. The sensitivity of the FQ-PCR assay reached 5 X 103 copies/ml in detecting BKV or CMV DNA. Control DNA verified the assay specifically detecting target DNA. The precision of the assay to quantif target DNA copies was acceptable (Intra-assay CV was 3.44% for BKV and 2.23% for CMV; Inter-assay CV was 4. 98% for BKV and 3.76% for CMV;). Of 480 samples, 130 samples (27. 08%) were CMV DNA positive, significantly higher than the BKV DNA positive (13.33%, 64/480, P<0.05). The positive BKV or CMV DNA was found to be associated with high concentrations of FK506 (P<0. 05). In conclusion, the developed real-time PCR assay for detecting both CMV and BKV DNA simultaneously was s high sensitive, precise and time-effectiveand could be applied in the monitoring of the CMV and BKV infection in the renal transplantation recipients.
Adolescent ; Adult ; Aged ; Conserved Sequence ; Cytomegalovirus ; genetics ; isolation & purification ; Cytomegalovirus Infections ; diagnosis ; virology ; DNA, Viral ; blood ; Female ; Humans ; Immunosuppressive Agents ; blood ; Kidney Transplantation ; adverse effects ; Male ; Middle Aged ; Polyomavirus ; genetics ; isolation & purification ; Polyomavirus Infections ; diagnosis ; virology ; Real-Time Polymerase Chain Reaction ; methods ; Reproducibility of Results ; Sensitivity and Specificity ; Species Specificity ; Tacrolimus ; blood ; Time Factors ; Tumor Virus Infections ; diagnosis ; virology ; Viral Load ; Young Adult

OBJECTIVETo experimentally investigate direct intramuscular gene transfer of nanoparticles encoding vascular endothelial growth factor for the treatment of peripheral artery disease.

METHODSThe human VEGF(165) cDNA was cloned into the eukaryotic expression vector PIRES2 under the control of cytomegalovirus promoter/enhancer. The recombinant gene was transferred into a rabbit model of chronic hindlimb ischemia by naked plasmid and nanoparticle respectively. Ischemia was induced in the hindlimb of New Zealand White rabbits by ligation of the distal external iliac artery and complete excision of the femoral artery and all its branches. At day 7 postoperation animals received VEGF(165) plasmid (10 intramuscular) or nanoparticle-VEGF(165) (8 intramuscular). With RT-PCR, immunohistochemistry analysis, and angiography, the expression and biological effects of VEGF(165) gene in experimental animals were investigated.

RESULTSTwo weeks after initiation of therapy, angiography showed that the transfer of VEGF(165) gene stimulated the formation of focal neovessels and established collateral circulation. The adductor muscle of ischemic limbs was histologically examined at day 14. Capillary density was increased among VEGF(165)-transfected rabbits, especially Nano-VEGF(165)-treated animals (Naked VEGF(165) plasmid = 50.18 per mm(2), Nano-VEGF(165) = 81.22 per mm(2), Control = 29.54 per mm(2), P < 0.05). RT-PCR showed that the transcription and expression of VEGF(165) gene in experimental group were significantly higher than those of control groups.

CONCLUSIONSIntramuscular administration of VEGF(165) induces collateral artery augmentation in the rabbit model of chronic limb ischemia. Nanoparticle can act as a vector to transfect specific gene and it will benefit gene transfer.

Angiography ; Animals ; Capsules ; Chronic Disease ; Disease Models, Animal ; Genetic Therapy ; methods ; Hindlimb ; blood supply ; Immunohistochemistry ; Ischemia ; genetics ; therapy ; Male ; Nanotechnology ; Particle Size ; Rabbits ; Reverse Transcriptase Polymerase Chain Reaction ; Treatment Outcome ; Vascular Endothelial Growth Factor A ; genetics ; therapeutic use

Objective To explore the experience and methods of neuro-endoscopy in minimally invasive treating intracranial diseases.Methods The surgery effect and complications of minimally invasive treatment under neuro-endoscopy on 52 patients with intracranial diseases,admitted to our hospital from October 2010 to March 2013,were retrospectively summarized; in these 52 patients,20 had pituitary adenoma,20 had hypertensive intracerebral hemorrhage,1 had intracranial hematoma followed arteriovenous malformation,2 had intracranial hematomas after brain injury,2 had cholesteatoma,2 had optic canal decompression,2 had cerebrospinal fluid rhinorrhea,1 had multi-separated subacute subdural hematoma and the last 2 had arachnoid cyst.Results Except for 1 patient having cholesteatoma appeared bleeding of the petrosalvein intraoperatively,which resulted in cerebellum hemorrhage following postoperative venous hemorrhagic infarction and unfavourable prognosis,the rest of the patients recovered well; in 20 patients having hypophysoma,15 (75％) achieved total resection of the pituitary adenoma,and no postoperative cerebrospinal fluid rhinorrhea occurred; in 23 cases of intracranial hematomas,22 were cleared satisfactorily,and the left one occurred rehemorrhage in one side but craniotomy hematoma removal was successfully performed again.The eyesight of 2 patients with optic canal decompression was improved obviously.The repair effects of 2 patients with cerebrospinal fluid leakage were very good without intracranial infection or recurrence.The 2 patients with arachnoid cyst recovered satisfactorily without other complications.One patient with multi-separated subacute subdural hematoma recovered with hematoma clear postoperative satisfactorily; and one patient with cholesteatoma postoperative recovered satisfactorily with tumor resection.Conclusion Neuro-endoscopy has many advantages such as exposed wide range,good deep lighting,minimally invasive and high efficiency in endoscopic transnasal transsphenoidal skull base surgery and intracranial hematomas removal.

To establish a metabonomic method based on high performance liquid chromatography-quadrupole-time of flight mass spectrometry (HPLC-Q-TOF/MS), in order to study the changes in serum metabolites of systemic lupus erythematosus (SLE) mice after treatment of Jieduquyuziyin prescription, the pathogenesis of SLE and mechanism of drug action. The orthogonal partial least squares (OPLS) was applied for the pattern recognition of experimental data, finding a significant difference in the control group, the SLE model group, the Jieduquyuziyin prescription-treated group and the prednisone acetate-treated group. According to the OPLS load diagram, 12 differential metabolites, including traumatic acid, PAF, 12 (S)-HEPE, 15(S)-HETrE and Hepoxilin B3 were identified by using accurate mass combined with MS/MS data After treatment with Jieduquyuziyin prescription, the relative contents of PAF, 12 (S)-HETE were close to the level of the control group. According to the analysis on metabolic pathway, SLE could cause significant changes in unsaturated fatty acid and amino acid metabolism pathway, while Jieduquyuziyin prescription has a effect in regulating disorder of unsaturated fatty acid metabolism pathway.
Amino Acids ; blood ; Animals ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Fatty Acids ; blood ; Female ; Humans ; Lupus Erythematosus, Systemic ; blood ; drug therapy ; Male ; Mass Spectrometry ; Mice ; Serum ; chemistry

The object of the research was to extract, purify and identify the type II collagen of Agkistrodon acutus. Type II collagen of A. acutus was extracted by enzyme decomposition method, and purified by ion exchange column chromatography. It was characterized by SDS-PAGE gel electrophoresis, ultraviolet spectrophotometry, infrared absorption spectroscopy and mass spectroscopy. The results showed that the size of C II was about 130 kDa. It absorbed at 223 nm. IR spectrum obtained showed that the triple helical domains of amino-acid sequences were characterized by the repetition of triplets Gly-X-Y. The MS spectrum graphically stated that C II extracted from cow and A. acutus have the similar peptides. The C II of A. acutus was obtained by extraction and purification. Appraisal analysis by SDS-PAGE, UV, IR and MS, C II of A. acutus was consistent with the standard C II of cow. It was proved that the extracted protein was C II.
Agkistrodon ; metabolism ; Animals ; Collagen Type II ; chemistry ; isolation & purification ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Mass Spectrometry ; Reptilian Proteins ; chemistry ; isolation & purification ; metabolism

BACKGROUNDChina began to implement the national medical and health system and public hospital reforms in 2009 and 2012, respectively. Anhui Province is one of the four pilot provinces, and the medical reform measures received wide attention nationwide. The effectiveness of the above reform needs to get attention. This study aimed to master the efficiency and productivity of county-level public hospitals based on the data envelopment analysis (DEA) model and Malmquist index in Anhui, China, and then provide improvement measures for the future hospital development.

METHODSWe chose 12 country-level hospitals based on geographical distribution and the economic development level in Anhui Province. Relevant data that were collected in the field and then sorted were provided by the administrative departments of the hospitals. DEA models were used to calculate the dynamic efficiency and Malmquist index factors for the 12 institutions.

RESULTSDuring 2010-2015, the overall average relative service efficiency of 12 county-level public hospitals was 0.926, and the number of hospitals achieved an effective DEA for each year from 2010 to 2015 was 4, 6, 7, 7, 6, and 8, respectively, as measured using DEA. During this same period, the average overall production efficiency was 0.983, and the total productivity factor had declined. The overall production efficiency of five hospitals was >1, and the rest are <1 between 2010 and 2015.

CONCLUSIONSIn 2010-2015, the relative service efficiency of 12 county-level public hospitals in Anhui Province showed a decreasing trend, and the service efficiency of each hospital changed. In the past 6 years, although some hospitals have been effective, the efficiency of the county-level public hospitals in Anhui Province has not improved significantly, and the total factor productivity has not been effectively improved. County-level public hospitals need to combine their own reality to find their own deficiencies.