Objective: To investigate the effect of curcumin (Cur) on cycle in HepG2 cells and the action of microtubule system for it. Methods: The proliferation inhibition of Cur on HepG2 cells was evaluated by MTT assay; The effect of Cur on the cycle of HepG2 cells was analyzed by flow cytometry; Confocal laser scanning microscopy (CLSM) was used to observe the changes of microtubule in HepG2 cells after treated with Cur; Western blotting was performed to determine the expression of α-tubulin; The in vitro effects of Cur on tubulin polymerization and depolymerization activities were studied. Results: Cur inhibited the proliferation of HepG2 cells in a dose- and time-dependent manner. With Cur concentration increasing, the proportion of HepG2 cells arrested in G2/M phase was also increased. CLSM showed that Cur could seriously destroy the cell microtubule structure and change the polymerization state of tubulin. Western blotting showed that the decrease of the expression level of α-tubulin was related to Cur concentration. Cur could also interfere the microtubule polymerization and depolymerization activities. Conclusion: Cur could inhibit the proliferation of HepG2 cells by destroying the microtubule structure and downregulating the expression of α-tubulin, and make HepG2 cells arrested in G2/M phase.

Objective:To develop a HPLC method for determination of Podophyllotoxin in two preparations.Methods:HPLC was carried out,using a Dikma diamonsil C18 column(4.6mm?150mm,5?m)with a column temperature at 25℃,the methanol-water as mobile phase(using gradient)with flowing rate 1.0 mL/min.The detection wavelength was at 292 nm.Results:The method had good linear relationship within the range of 11.8-590?g/mL for Podophyllotoxin,with the correlation coefficients 0.9999.The results of the average recovery of Youdi liniment and Podophyllotoxin tincture were 99.38%,99.62%.Conclusion:The rapid,accurate and reproducible RP-HPLC method was useful for quantification of Podophyllotoxin in Youdi Liniment and Podophyllotoxin Tincture.

Objective To compare the feasibility and efficacy of intervertebral wedge osteotomy and pedicle subtraction osteotomy (PSO),vertebral column resection (VCR),Smith-Petersen osteotomy (SPO) for the treatment of severe kyphosis and scoliosis.Methods The data of 38 cases of severe kyphosis and kyphoscoliosis were retrospectively analyzed from January 2010 to February 2016,including 22 males and 16 females.According to the osteotomy mode,PSO,SPO,VCR and intervertebral disc wedge osteotomy were used to collect the average number of fixed phases,volume of bleeding,length of stay,length of hospital stay,improvement of main cobb angle,improvement of ODI score,and Frankel classification to evaluate the efficacy.Results There were no significant differences in the overall operative time between the four groups.The average number of fixation in 18 patients with SPO was (9.4±3.9) segments,the blood loss was (3 000±410) ml,the average Cobb angle was improved by 55.3％± 9.5％,the average postoperative hospitalization was (14.6±4.9) days,the improvement rate of ODI was 42.1％±7.4％,all the patients were improved to Frankel E;The average number of fixation in 5 patients with PSO was (7.6± 1.5) segments,the blood loss was (4 360± 1 161) ml,the average Cobb angle was improved by 58.9％ ± 15.1％,the average postoperative hospitalization was (18.2±7.0) days,the improvement rate of ODI was 41.3％±9.6％.One Frankel C patient was improved to Frankel D,others remained to be Frankel E as the same as pre-operation;The average number of fixation in 4 patients with VCR was (6.2±2.6) segments,the blood loss was (3 750+ 1 848) ml,the average Cobb angle was improved by 83.9％± 10.9％,the average postoperative hospitalization was (21±7.2) days,the improvement rate of ODI was 39.6％± 18.1％.Three Frankel D patients were improved to Frankel E and one Frankel C patient was improved to Frankel D;The average number of fixation in 11 patients with IWO was (7.1 ± 2.7) segments,the blood loss was (2855±1046) ml,the average Cobb angle was improved by 59.6％±22.05％,the average postoperative hospitalization was (13.5±2.7) days,the improvement rate of ODI was 51.3％±8.3％.One Frankel C patient was improved to Frankel D,eight Frankel D patients were improved to Frankel E,other patients remained to be Frankel E;The mean follow-up time was 25.2 months in 11 patients underwent intervertebral wedge osteotomy.All the patients had successful spinal fusion and no failure of internal fixation.Conclusion Intervertebral wedge osteotomy for the treatment of scoliosis and kyphosis could reduce surgical injury to obtain good biomechanics and surgical result.

Objective - - （BCL2L2）- （ ） To investigate the differential expression of the fusion gene BCL 2 like protein 2 poly A （PABPN1） （ ） binding protein nuclear 1 induced by sodium arsenite SA and its methylated metabolites in 16HBE cells and the Methods ） ， related mechanism. i The 16HBE cells exposed to SA at concentrations of 1.5 3.0 and 4.5 µmol/L were set as -， - - low medium and high dose arsenic exposure groups. The 16HBE cells exposed to 4.5 µmol/L monomethylarsonic acid （ ）， （ ） ， MMA dimethylarsonic acid DMA and SA were set as MMA group DMA group and SA group. The 16HBE cells without ， BCL2L2-PABPN1 toxic stimulation were set as control group. After the cells were cultured for 48 hours the expression of was - （ - ） ） （ ） detected by quantitative real time polymerase chain reaction qRT PCR . ii Two small interfering RNA siRNA silencing 基金项目：国家自然科学基金（ ）； 年云南省科技厅昆明医科大学应用基础研究联合专项面上项目 82160607 2021 （ ） 202101AY070001-054 作者简介：施雅（ —），女，在读大学本科生，主要从事劳动卫生与环境卫生学研究；尹锦瑶（ —），女，在读劳动卫生与环境卫 2001 1995 生学硕士研究生，主要从事劳动卫生与环境卫生学研究；施雅和尹锦瑶为共同第一作者 通讯作者：何越峰教授，博士研究生导师，- ： E mail heyuefeng@kmmu.edu.cn中国职业医学 年 月第 卷第 期 ， ， ， · · 2022 10 49 5 Chin Occup Med October 2022 Vol.49 No.5 523 BCL2L2-PABPN1， - fragments were designed and transfected into 16HBE cells to knockdown which were set as siRNA 1 group - - BCL2L2-PABPN1 and siRNA 2 group. Non transfected control group without knockdown of transfection was set up. After ， BCL2L2-PABPN1 - culturing for 48 hours the expression level of in the three groups of cells was detected by qRT PCR. The cell - survival rate and early apoptosis rate were detected by MTS method and JC 1 mitochondrial membrane potential detection ， （ ） ， method respectively. The apoptosis was detected by Hoechest33342/propidium iodide PI double staining and the expression - Results ） level of P53 signaling pathway related proteins was detected by Western blotting. i The relative expression of BCL2L2-PABPN1 （P ） BCL2L2- in 16HBE cells increased with the increasing SA doses <0.01 . The relative expression of PABPN1 - ， - - in high dose arsenic exposure was higher than that in control group low dose and medium dose arsenic exposure （ P ） BCL2L2-PABPN1 ， groups all <0.05 . The relative expression of in SA group was higher than those in control group MMA （ P ） BCL2L2-PABPN1 group and DMA group all <0.05 . The relative expression of showed no significant difference between ， （ P ） ） BCL2L2-PABPN1 control group MMA group and DMA group all >0.05 . ii The relative expression levels of and cell - - - （ P ） survival rate in siRNA 1 group and siRNA 2 group were lower than those in non transfected control group all <0.05 . ， （P ） However there was no significant difference in the early apoptosis rate among the three groups >0.05 . The results of - Hoechest33342/PI double staining showed that the number of nuclear shrinkage and early apoptotic cells in siRNA 1 group and - - ， - siRNA 2 group was higher than that in non transfected control group. The relative protein expression levels of P53 phospho ， - - ， - - （ P ） p53 BCL 2 associated death promoter P21 and cytochrome C in siRNA 1 group and siRNA 2 group were higher all <0.05 , - - P and the relative protein expression levels of P53 up regulated modulator of apoptosis were lower (all <0.05), when compared - Conclusion with the non transfected control group. SA may block the apoptosis of 16HBE cells by inducing the expression of BCL2L2-PABPN1 fusion gene . The mechanism may be related to the activation of P53 signaling pathway. The SA methylated BCL2L2-PABPN1 BCL2L2-PABPN1 - metabolites MMD and DMA had no effect on the expression of . may affect anti apoptosis BCL2L2 PABPN1 through affecting the synergistic effect of and genes.

OBJECTIVETo explore the predictive value of sperm chromatin integrity test (SCIT) in assisted reproductive technology (ART) by analyzing the relationship of sperm chromatin integrity (SCI) with the outcomes of IVF-ET and ICSI.

METHODSSperm chromatin structure assay (SCSA) was performed to test SCI in 187 ART cycles, and the results were expressed as DNA fragmentation index (DFI). According to the level of DFI, the 187 cycles were allocated to a high DFI group (DFI > or = 30% ) and a low DFI group (DFI < 30%), each of which was again divided into an IVF and an ICSI subgroup. Comparisons were made between the IVF and ICSI subgroups of the high and low DFI groups in the fertilization rate, cleavage rate, embryo quality, and clinical pregnancy rate.

RESULTSThe clinical pregnancy rate of ICSI was significantly higher than that of IVF in the high DFI group, while the clinical outcomes showed no significant differences between the high and low DFI groups in either the IVF or the ICSI subgroup.

CONCLUSIONSperm DNA damage affects the outcome of ART, and therefore SCIT can be used as a supplementary option to standard semen analysis in choosing the method for ART.

Adult ; Chromatin ; DNA Fragmentation ; Female ; Fertilization in Vitro ; Humans ; Male ; Predictive Value of Tests ; Pregnancy ; Pregnancy Rate ; Semen Analysis ; Sperm Injections, Intracytoplasmic ; Spermatozoa ; cytology

OBJECTIVETo investigate the clinical effects of the combined therapy of the Chinese medicine Compound Xuanju Capsule and vitamin E on sperm chromatin damage in idiopathic oligoasthenospermia.

METHODSWe assigned 50 infertile men with seminal abnormality to a control group (n = 26) and a trial group (n = 24) to receive vitamin E and the combined therapy of Compound Xuanju Capsule plus vitamin E, respectively, both treated for 3 months. Before and after the treatment, we detected semen routine parameters and sperm DNA fragmentation indexes (DFI) by computer aided semen analysis (CASA) and sperm chromatin structure assay (SCSA), and compared them between the two groups.

RESULTSThere was no obvious difference between the percentage of progressively motile sperm in the trial group and that in the control group (21.55 +/- 8.68 vs 21.47 +/- 11.53, P > 0.05). The trial group showed a significantly decreased sperm DFI after medication as compared with pre-medication (29.57 +/- 12.19 vs 34.09 +/- 10.32, P < 0.05).

CONCLUSIONThe combined therapy of Compound Xuanju Capsule and vitamin E can effectively improve seminal quality and reduce sperm chromatin damage in infertile men with idiopathic oligoasthenospermia.

Adult ; Capsules ; Chromatin ; drug effects ; DNA Damage ; drug effects ; DNA Fragmentation ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Humans ; Infertility, Male ; drug therapy ; genetics ; Male ; Spermatozoa ; drug effects ; Vitamin E ; pharmacology ; therapeutic use ; Young Adult

AIM: To investigate the antitumor activity and mechanism of Paecilomyces tenuipes polysaccharide(PTPS).METHODS: PTPS-I was obtained by water extraction and alcohol precipitation,and purified by DEAE-cellulose and Sephadex G-100 chromatography.Human erythroleukemia cell line K562,laryngocarcinoma cell line Hep2 and hepatic carcinoma cell line SMMC-7721were co-cultured with PTPS-I or the conditioned medium which prepared with PTPS-I-stimulated human mononuclear cells(PTPS-I-MNC-CM),and the proliferation of tumor cells was determined.The cell counting kit-8(CCK-8) was used to determine the proliferation of MNCs.The FQ-RT-PCR was applied to investigate the expression of TNF-? and IL-6 mRNA in MNCs.RESULTS: PTPS-I-MNC-CM inhibited the proliferation of K562,Hep2 and SMMC-7721 cells in vitro(P

Objective To investigate the differences between the sequences of papA and papG of UPECA030 strain and the related genes, to better understand the genetic variation of UPEC4030 papA and its combination forms with papG so as to identify if it was a new genotype. Methods Cloning and sequencing methods were used to analyze the sequences of papG and papA of UPEC4030 strain and to compare their related sequences. Results Through sequence analysis of papA, it was revealed that there was a 722 bp gene,encoding 192 amino acid polypeptide. The overall homology of the papa genes between UPEC4030 and the standard strains of ten F types were 36.11%-77.95 % and 22.20%-78.34% at nueleotide and deduced amino acid levels. Homology between the sequences of reverse primers and the corresponding sequence of UPEC4030 papA was 10.00%-66.67%. The results confirmed that UPECA030 strain contained a novel papA variant. Through sequence analysis of UPEC4030 papG, we revealed a 1100 bp gene, encoding 337 amino acid polypeptide. The homology of the papG genes between UPEC4030 and UPEC IA2, the standard strain, was 99.00 % at nucleotide level and 99.11% at deduced amino acid and UPEC4030 strain carried class I] genotype of papG. Conclusion UPEC4030 strain contained an unknown papA variant or the new genotype and carried class II genotype of papG. The pathogenic mechanism and epidemiology call for further study.

AIMTo establish an HPLC-MS method for determination of octreotide in plasma and study the relative bioavailability of domestic and imported octreotide injections.

METHODSOctreotide in plasma samples were extracted with a Waters solid-phase extraction mini column. HPLC-MS was carried out using a Waters Xetrra C18 column and a mobile phase consisting of CH3 OH-1% HAc (80 : 20), the flow rate was 0.2 mL x min(-1), and the internal standard was 6, 7, 4'-OH-isoflavone, the SIR ions for quantification were m/z 1 014.4 for octreotide and m/z 317.6 for internal standard. A single dose of 200 microg of domestic or imported preparations was intramuscularly given to 18 healthy volunteers in a randomized crossover study. Octreotide concentration in plasma was determined by LC-MS method. The pharmacokinetics and bioavailability were studied.

RESULTSThe regressive curve was linear (r = 0.9997) within the range of 0.5 - 40 microg x L(-1) for octreotide. The pharmacokinetics parameters of domestic and imported injection were reply to one compartment model. The mean C(max) were (19 +/- 10) microg x L(-1) and (19 +/- 11) microg x L(-1), T(max) were (0.50 +/- 0.15) h and (0.52 +/- 0.20) h, T1/2 were (1.5 +/- 0.8) h and (1.5 +/- 0.8) h, AUC(0-7 h) were (50 +/- 25) h x microg x L(-1) and (50 +/- 25) h x microg x L(-1), respectively. The relative bioavailability of domestic to imported injection was 101% +/- 10%.

CONCLUSIONThe method is accurat and sensible for assay of plasma octreotide concentration. The results of statistics showed the two preparations were bioequivalent.

Area Under Curve ; Biological Availability ; Chromatography, High Pressure Liquid ; Cross-Over Studies ; Gas Chromatography-Mass Spectrometry ; Humans ; Injections, Intramuscular ; Octreotide ; administration & dosage ; blood ; pharmacokinetics

Objective: : To study the physiochemical characteristics of podophyllotoxin (PPT) conjugated stearic acid grafted chitosan oligosaccharide micelle (PPT-CSO-SA), and evaluate the ability of the potential antineoplastic effects against glioma cells. Methods: : PPT-CSO-SA was prepared by a dialysis method. The quality of PPT-CSO-SA including micellar size, zeta potential, drug encapsulation efficiency and drug release profiles was evaluated. Glioma cells were cultured and treated with PPT and PPT-CSO-SA. The ability of glioma cells to uptake PPT-CSO-SA was observed. The proliferation of glioma cells was determined by 3-[4, 5-dimethyl-2-thiazolyl]-2, 5-diphenyl-2H-tetrazolium bromide (MTT) assay. The apoptosis and morphology of U251 cells were observed by 4’,6-Diamidino-2-phenylindole dihydrochloride (DAPI) dye staining. Cell cycle analysis was performed by flow cytometry. The migration ability of U251 cells was determined by wound healing test. Results: : PPT-CSO-SA had nano-level particle size and sustained release property. The encapsulation efficiency of drug reached a high level. The cellular uptake percentage of PPT in glioma cells was lower than that of PPT-CSO-SA (p<0.05). The inhibitory effect of PPT-CSO-SA on glioma cells proliferation was significantly stronger than that of PPT (p<0.05). The morphologic change of apoptosis cell such as shrinkage, karyorrhexis and karyopyknosis were observed. The percentage of U251 cells in G2/M phase increased significantly in the PPT-CSO-SA group compared with PPT group (p<0.05). Compared with the PPT group, the cell migration ability of the PPT-CSO-SA group was significantly inhibited after 12 and 24 hours (p<0.05). Conclusion : PPT-CSO-SA can effectively enhance the glioma cellular uptake of drugs, inhibit glioma cells proliferation and migration, induce G2/M phase arrest of them, and promote their apoptosis. It may be a promising anti-glioma nano-drug.