Objective KIF18A is a protein that has close relation with mitotic regulation and tumor development.This study aimed to establish KIF18A protein expression system in baculovirus,which may help to realize high efficient synthesis of KIF1 8A protein in vitro.Methods Transfer vector was constructed with molecular cloning method,and recombinant baculovirus were obtained through gene transposition.KIF18A protein was expressed by transforming recombinant baculovirus into infected insect Sf-9 cells to realize the synthesis efficiency.Results It was confirmed by DNA sequencing,microscopic observation and Western Blot that the KIF18A protein expression system in baculovirus was successfully established.Conclusions Conditions for transfer vector and recombinant virus transfection are defined,and high efficient KIF18A protein baculovirus expression system are successfully constructed.

Objective To purify pre-ribosome and ribosome of mammalian ceils using continuous sucrose density gradient ultracentrifugation.Methods Continuous sucrose density gradient was established by ultracentrifugation,and the continuous sucrose density gradient of 10％-30％ and 10％-45％ were used to extract the pre-ribosome and ribosome in mammalian cells,respectively.The mammalian cell lysis buffer was added to the established continuous sucrose density gradient.Pre-ribosome and ribosome with different sedimentation coefficients were collected and the A260 absorbance of each sample was measured.Proteins of each sample were extracted to detect the large subunit protein,RPL15 by Western Blot.Results Large subunit ribosomal protein RPL15 exists on 60S of the pre-ribosome,and also on 60S,80S and polyribosome of mature ribosome.Conclusions The continuous sucrose density gradient,which is established by the swing-out rotor,can be used to isolate the pre-ribosome and ribosome of mammalian cells rapidly.This method has the advantages of good separation effect and simple operation,which provides a good method for rapid and large amount preparation and separation of various kinds of ribosomes.

Objective To investigate the process that chromosome kinesin KIF4A promote cisplatin resistance in lung cancer cells.Methods Reverse transcription PCR (RT-PCR) and Western Blot experiments were performed to analyze the expression of KIF4A in lung cancer cells A549 and cisplatin (DDP) resistant cells A549/ DDP.Cell transfection, RNA interference (RNAi) experiments and thiazolyl blue tetrazolium bromide (MTT) assays were carried out to examine cell proliferation of A549 cells with overexpression of exogenous KIF4A and A549/DDP cells with depletion of endogenous KIF4A after cisplatin treatment.Results Expression of KIF4A in A549/DDP cells was higher than that in A549 cells.With overexpression of exogenous KIF4A, A549 cells displayed drug resistance to cisplatin.On the contrary, depletion of endogenous KIF4A in A549/DDP cells resulted in cisplatin sensitivity.Conclusions Chromosome kinesin KIF4A involves in the regulation of cisplatin resistance in lung cancer cells and KIF4A may be a potential and effective new biological target for treatment of lung cancer cisplatin resistance.

Objective:To detect the expression and methylation of TIA1 in breast cancer and to study its correlation with multi-slice spiral CT signs.Methods:50 patients with breast cancer were collected from Feb.2019 to Mar. 2020. The expression levels of TIA1 in breast cancer tissues and in peritumoral tissues were estimated by quantitative real-time polymerase chain reaction. Bioinformatics software MethPrimer was used for predicting TIA1 promotor and confirmed the existence of cPG island. Methylation-specific PCR (MSP) was performed to detect TIA1 DNA promoter methylation. All patients were examined by multi-slice CT. CT images were analyzed through observing the tumor size, shape, calcification area, lymph node metastasis and margin. The correlation between CT signs and TIA1 methylation status was further analyzed.Results:The expression levels of TIA1 in breast cancer tissues were lower than in peritumoral tissues (0.50±0.12, 0.95±0.10, P=0.00) , while TIA1 DNA promoter methylation rate was higher than in peritumoral tissues (64%, 42%, χ2=4.86, P<0.05) .There were no significant differences in TIA1 DNA promoter methylation rate among patients with different tumor shape and micro calcifications. TIA1 DNA promoter methylation rate in patients with mass diameter≥2 cm were significantly higher than those in patients with mass diameter<2 cm (78.57%, 45.45%, P<0.05) , and TIA1 DNA promoter methylation rate in patients with lymph node metastasis was higher than those without lymph node metastasis (79.17%, 50%, P<0.05) . TIA1 DNA promoter methylation rate in patients with burr at edge of mass was higher than those without burr at edge of mass (80.77%, 45.83%, P<0.05) . Conclusion:There is a correlation between CT imaging signs and TIA1 DNA promoter methylation rate in patients with breast cancer, which can provide more reference for the judgment of malignant degree and prognosis of patients with breast cancer.

Objective To investigate the clinical significance and changes of serum complement in lipid metabolism disorder in patients with fatty liver disease.Methods One hundred and forty patients with FLD from October 2015 to May 2017 were included in the study,in addition,120 patients with hyperlipidemia(the hyperlipidemia group)and 130 healthy subjects(the control group)in the same period were enrolled as controls.The differences in serum lipid,liver function enzymology,immunoglobulin,serum complement among the three groups were compared.Results There were significant differences in the levels of TC,TG,HDL-C,LDL-C,apoA and apoB among the three groups(TC:(5.7±1.6)mmol/L vs.(4.2±1.0)mmol/L vs.(3.5±1.1) mmol/L,F=105.01,P<0.05;TG:(2.8± 0.6)mmol/L vs.(1.5 ± 0.3)mmol/L vs.(1.1 ± 0.2)mmol/L,F=628.46,P<0.05;HDL-C:(1.2±0.3)mmol/L vs.(1.5±0.3)mmol/L vs.(1.8±0.4)mmol/L,F=107.10, P<0.05;LDL-C:(3.6±0.9)mmol/L vs.(3.0±0.8)mmol/L vs.(2.2±0.6)mmol/L,F=109.07,P<0.05;apoA:(1.0±0.2)g/L vs.(1.2±0.2)g/L vs.(1.4±0.3)g/L,F=95.20,P<0.05;apoB:(1.1±0.2)g/L vs.(0.9±0.2)g/L vs.(0.8±0.2)g/L,F=79.04,P<0.05).The levels of TC,TG,LDL-C and apoB in the FLD group were significantly higher than those in the hyperlipidemia group and the control group.The levels of HDL-C and apoA in the FLD groups were significantly lower than those in the hyperlipidemia group and control group.The levels of TC,TG,LDL-C and apoB in the hyperlipidemia group were significantly higher than those in the control group.The levels of HDL-C and apoA in the hyperlipidemia group were significantly lower than those in the control group.There were significant differences in the levels of GGT,ALT,AST,IgG and IgM among the three groups(GGT:(77.4±15.3)U/L vs.(43.3±10.6)U/L vs.(25.5±8.2)U/L,F=668.12,P<0.05;ALT:(61.5±18.8)U/L vs.(35.7±11.2)U/L vs.(18.9±5.4)U/L,F=355.67,P<0.05;AST:(55.3±12.2)U/L vs.(32.4±12.5)U/L vs.(14.4±4.7)U/L,F=521.80,P<0.05;IgG:(15.7±3.9)g/L vs.(11.6±3.2)g/vs.,(8.5±2.6)g/L,F=162.34,P<0.05;IgM:(1.9±0.6)g/L vs.(1.2±0.4)g/L vs.(0.8±0.3)g/L,F=201.38,P<0.05).The levels of GGT,ALT,AST,IgG and IgM in the FLD group were significantly higher than those in the hyperlipidemia group and the control group.The levels of GGT,ALT,AST,IgG and IgM in the hyperlipidemia group were significantly higher than those in the control group.There were significant differences in the levels of C3,C5,ASP and BF among the mild,moderate and severe fatty liver patients(C3:(2.1±0.4) g/L vs.(1.8±0.3)g/L vs.(1.0±0.2)g/L,F=436.37,P<0.05;C5:(92.3±10.7)mg/L vs.(71.8±8.8) mg/L vs.(58.9±6.5)mg/L,F=486.09,P<0.05; ASP:(51.4±6.8)nmol/L vs.(42.5±4.4)nmol/L vs.(32.8±5.2)nmol/L,F=369.29,P<0.05;BF:(0.48±0.13)g/L vs.(0.34±0.09)g/L vs.(0.23±0.04) g/L,F=233.39,P<0.05).The levels of C3,C5,ASP and BF in the FLD group were significantly higher than those in the hyperlipidemia group and the control group.The levels of C3,C5,ASP and BF in the hyperlipidemia group were significantly higher than those in the control group.There were significant differences in the levels of C3,C5,ASP and BF among patients with mild,moderate and severe fatty liver disease(C3:(1.8±0.3)g/L vs.(2.1±0.4)g/L vs.(2.5±0.4)g/L,F=30.85,P<0.05;C5:(80.5±9.6)mg/L vs.(92.3±10.5)mg/L vs.(100.7±8.)mg/L,F=39.39,P<0.05; ASP:(42.4±6.3)nmol/L vs.(52.8±5.7)nmol/L vs.(61.9±5.6) nmol/L,F=98.19,P<0.05;BF:(0.33±0.12)g/L vs.(0.45±0.11)g/L vs.(0.57±0.09)g/L,F=41.26,P<0.05).The levels of C3,C5,ASP and BF in the mild FLD patients were significantly lower than those in moderate and severe FLD patients.The levels of C3,C5,ASP and BF in the moderate FLD patients were significantly lower than those in severe FLD patients.Conclusion The detection of C3,C5,ASP and BF levels based on routine testes has important clinical value for the assessment of the condition,the treatment and the prognosis of FLD patients.

Objective To investigate the relationship between the environmental tobacco smoke (ETS) and lung cancer by Meta-analysis. Methods We used“lung cancer/lung neoplasm”,“non-smoking/non-smoker”,“China/Chinese”,“case-control/case control”,“risk factor”,“environmental tobacco smoke/passive smoking”as key words, to search papers in databases including Chinese BioMedical Literature (CBM), China National Knowledge Internet (CNKI), Wanfang, Vip Citation Databases (VIP), PubMed and Web of Science databases, and collected the case-control studies on ETS and lung cancer among Chinese non-smokers from January 1999 to December 2013. A total of 129 research papers were collected. RevMan 5.2 software was used to calculate combined odds radio (OR) and 95%CI. Results Qualified 18 literatures were included, total cases 6 145 and controls 8 132. Consolidated results showed that ETS exposure could increase the risk of lung cancer, combined OR(95%CI)=1.52(1.42-1.64). Stratified analysis showed that ETS exposure was found to be significantly associated with an increasing risk of the lung cancer on non-smoking women and men, and combined OR(95%CI) were 1.58(1.42-1.75) and 1.34(1.08-1.65), respectively; the ETS exposure from family or the working environment could increase the risk of lung cancer, and combined OR(95%CI) were 1.48(1.20-1.82) and 1.38(1.13-1.69) respectively;childhood exposure and adult exposure were no significant statistical significance, and combined OR(95%CI) were 1.37(0.98-1.91), and 1.34(0.97-1.85) respectively. Conclusion Environmental tobacco smoke exposure was a significant risk factor of lung cancer among non-smokers in China.

Objective To investigate the relationship between the environmental tobacco smoke (ETS) and lung cancer by Meta-analysis. Methods We used“lung cancer/lung neoplasm”,“non-smoking/non-smoker”,“China/Chinese”,“case-control/case control”,“risk factor”,“environmental tobacco smoke/passive smoking”as key words, to search papers in databases including Chinese BioMedical Literature (CBM), China National Knowledge Internet (CNKI), Wanfang, Vip Citation Databases (VIP), PubMed and Web of Science databases, and collected the case-control studies on ETS and lung cancer among Chinese non-smokers from January 1999 to December 2013. A total of 129 research papers were collected. RevMan 5.2 software was used to calculate combined odds radio (OR) and 95%CI. Results Qualified 18 literatures were included, total cases 6 145 and controls 8 132. Consolidated results showed that ETS exposure could increase the risk of lung cancer, combined OR(95%CI)=1.52(1.42-1.64). Stratified analysis showed that ETS exposure was found to be significantly associated with an increasing risk of the lung cancer on non-smoking women and men, and combined OR(95%CI) were 1.58(1.42-1.75) and 1.34(1.08-1.65), respectively; the ETS exposure from family or the working environment could increase the risk of lung cancer, and combined OR(95%CI) were 1.48(1.20-1.82) and 1.38(1.13-1.69) respectively;childhood exposure and adult exposure were no significant statistical significance, and combined OR(95%CI) were 1.37(0.98-1.91), and 1.34(0.97-1.85) respectively. Conclusion Environmental tobacco smoke exposure was a significant risk factor of lung cancer among non-smokers in China.

OBJECTIVETo investigate the relationship between the environmental tobacco smoke (ETS) and lung cancer by Meta-analysis.

METHODSWe used "lung cancer/lung neoplasm", "non-smoking/non-smoker", "China/Chinese", "case-control/case control", "risk factor", "environmental tobacco smoke/passive smoking" as key words, to search papers in databases including Chinese BioMedical Literature (CBM), China National Knowledge Internet (CNKI), Wanfang, Vip Citation Databases (VIP), PubMed and Web of Science databases, and collected the case-control studies on ETS and lung cancer among Chinese non-smokers from January 1999 to December 2013. A total of 129 research papers were collected. RevMan 5.2 software was used to calculate combined odds radio (OR) and 95% CI.

RESULTSQualified 18 literatures were included, total cases 6 145 and controls 8 132. Consolidated results showed that ETS exposure could increase the risk of lung cancer, combined OR (95% CI) = 1.52 (1.42-1.64). Stratified analysis showed that ETS exposure was found to be significantly associated with an increasing risk of the lung cancer on non-smoking women and men, and combined OR (95% CI) were 1.58 (1.42-1.75) and 1.34 (1.08-1.65), respectively; the ETS exposure from family or the working environment could increase the risk of lung cancer, and combined OR (95% CI) were 1.48 (1.20-1.82) and 1.38 (1.13-1.69) respectively; childhood exposure and adult exposure were no significant statistical significance, and combined OR (95% CI) were 1.37 (0.98-1.91), and 1.34 (0.97-1.85) respectively.

CONCLUSIONEnvironmental tobacco smoke exposure was a significant risk factor of lung cancer among non-smokers in China.

ObjectiveTo observe the clinical efficacy of tonifying kidney and replenishing essence on asthenozoospermia patients with the syndrome of kidney essence deficiency and the effects of this method on the adenosine 5′-monophosphate-activated protein kinase （AMPK）/mammalian target of rapamycin complex 1 （mTORC1） signaling pathway. MethodsSeventy-two eligible asthenozoospermia patients with the syndrome of kidney essence deficiency treated in the Affiliated Hospital of Chengdu University of Traditional Chinese Medicine from February 2023 to January 2024 were selected and randomly assigned into an observation group and a control group， with 36 patients in each group. The observation group received oral administration of Guilu Tianjing capsules， while the control group received oral administration of L-carnitine oral solution. The treatment course lasted for 4 weeks in both groups. The observed indicators included sperm progressive motility rate （PR）， total sperm motility （PR+NP）， percentage of normal mitochondrial membrane potential （MMP）， and traditional Chinese medicine （TCM） symptom scores before and after treatment in both groups. A three-month follow-up was instituted to record the conception status of the patients’ spouses. Additionally， eight patients were randomly selected from the eligible patients in the observation group， and four healthy males with normal semen routine examination results were included as the control group for the determination of protein expression. Western blotting was conducted to assess the expression of AMPK， phosphorylated （p）-AMPK， regulatory-associated protein of mTOR （RAPTOR） and p-RAPTOR， and PTEN-induced putative kinase 1 （PINK1） in sperms from the observation group before and after treatment， as well as in the sperms of the control group. ResultsThe pregnancy rate of spouses in the observation group was 9.09% （3/33）， which was higher than that （3.33%， 1/30） in the control group. The total response rate was 84.8% （28/33） in the observation group and 66.7% （20/30） in the control group， with no statistically significant difference. After treatment， both groups were improved considering PR， PR+NP， MMP， and TCM symptom scores （P<0.01）. Moreover， the observation group exhibited more pronounced decreases in TCM symptom scores than the control group （P<0.05）， while the changes in PR， PR+NP， and MMP showed no statistical significance between groups. Compared with the control group， the asthenozoospermia group exhibited upregulations in phosphorylation levels of AMPK and RAPTOR and protein level of PINK （P<0.01）. The administration of Guilu Tianjing Capsules led to downregulations in the phosphorylation levels of AMPK and RAPTOR and protein level of PINK1 （P<0.01）. However， the protein levels of AMPK and RAPTOR demonstrated no significant difference between before and after treatment. During the study period， neither group of patients exhibited any notable adverse reactions. ConclusionGuilu Tianjing capsules can enhance the sperm motility and percentage of normal mitochondrial membrane potential in asthenozoospermia patients with the syndrome of kidney essence deficiency by downregulating the AMPK/mTORC1 signaling pathway， lowering the protein level of PINK1， and inhibiting excessive activation of mitophagy.

ObjectiveTo observe the clinical efficacy of tonifying kidney and replenishing essence on asthenozoospermia patients with the syndrome of kidney essence deficiency and the effects of this method on the adenosine 5′-monophosphate-activated protein kinase （AMPK）/mammalian target of rapamycin complex 1 （mTORC1） signaling pathway. MethodsSeventy-two eligible asthenozoospermia patients with the syndrome of kidney essence deficiency treated in the Affiliated Hospital of Chengdu University of Traditional Chinese Medicine from February 2023 to January 2024 were selected and randomly assigned into an observation group and a control group， with 36 patients in each group. The observation group received oral administration of Guilu Tianjing capsules， while the control group received oral administration of L-carnitine oral solution. The treatment course lasted for 4 weeks in both groups. The observed indicators included sperm progressive motility rate （PR）， total sperm motility （PR+NP）， percentage of normal mitochondrial membrane potential （MMP）， and traditional Chinese medicine （TCM） symptom scores before and after treatment in both groups. A three-month follow-up was instituted to record the conception status of the patients’ spouses. Additionally， eight patients were randomly selected from the eligible patients in the observation group， and four healthy males with normal semen routine examination results were included as the control group for the determination of protein expression. Western blotting was conducted to assess the expression of AMPK， phosphorylated （p）-AMPK， regulatory-associated protein of mTOR （RAPTOR） and p-RAPTOR， and PTEN-induced putative kinase 1 （PINK1） in sperms from the observation group before and after treatment， as well as in the sperms of the control group. ResultsThe pregnancy rate of spouses in the observation group was 9.09% （3/33）， which was higher than that （3.33%， 1/30） in the control group. The total response rate was 84.8% （28/33） in the observation group and 66.7% （20/30） in the control group， with no statistically significant difference. After treatment， both groups were improved considering PR， PR+NP， MMP， and TCM symptom scores （P<0.01）. Moreover， the observation group exhibited more pronounced decreases in TCM symptom scores than the control group （P<0.05）， while the changes in PR， PR+NP， and MMP showed no statistical significance between groups. Compared with the control group， the asthenozoospermia group exhibited upregulations in phosphorylation levels of AMPK and RAPTOR and protein level of PINK （P<0.01）. The administration of Guilu Tianjing Capsules led to downregulations in the phosphorylation levels of AMPK and RAPTOR and protein level of PINK1 （P<0.01）. However， the protein levels of AMPK and RAPTOR demonstrated no significant difference between before and after treatment. During the study period， neither group of patients exhibited any notable adverse reactions. ConclusionGuilu Tianjing capsules can enhance the sperm motility and percentage of normal mitochondrial membrane potential in asthenozoospermia patients with the syndrome of kidney essence deficiency by downregulating the AMPK/mTORC1 signaling pathway， lowering the protein level of PINK1， and inhibiting excessive activation of mitophagy.