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Objective:To investigate the incidence and risk factors of renal injury in human immunodeficiency virus (HIV) infection/acquired immunodeficiency syndrome (AIDS) patients with poor immune reconstitution.Methods:The HIV infection/AIDS patients with poor immune reconstitution who were visited Second Department of Infection of Hangzhou Xixi Hospital from January to December 2021 were enrolled. The clinical data and laboratory examinations of the patients were collected, and the relevant risk factors were analyzed by logistic regression.Results:Among 303 HIV infection/AIDS patients with poor immune reconstitution, 59(19.5%) patients had renal injury. Logistic regression analysis showed that hypertension (odds ratio ( OR)=0.200, 95% confidence interval (95% CI) 0.065 to 0.618, P=0.005), taking tenofovir ( OR=0.275, 95% CI 0.130 to 0.580, P=0.001), hypoproteinemia ( OR=1.045, 95% CI 1.006 to 1.086, P=0.022), and low CD4 + T lymphocytes level ( OR=1.009, 95% CI 1.003 to 1.014, P=0.001) were risk factors for renal injury. Conclusions:The incidence of renal injury in HIV infection/AIDS patients with poor immune reconstitution is high. Hypertension, taking tenofovir, hypoproteinemia, and low CD4 + T lymphocytes level are risk factors for renal injury in patients.
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Aim To construct Flp-In CHO cell line(CYP2A13-CHO)stably expressing cytochrome P450 family 2 subfamily A member 13(CYP2A13)and Flp-In CHO cell line(CYP2A13-POR-CHO)stably co-expressing CYP2A13 and cytochrome P450 oxidoreductase(POR), from which a cell line with better metabolic activity is selected.Method In our previous study, we had constructed a Flp-In CHO cell line(POR-Flp-In CHO)stably expressing POR using lentiviral vector.The recombinant plasmids of pcDNA5/FRT-CYP2A13 were constructed and transfected into Flp-In CHO cells and POR-Flp-In CHO cells through LipofectamineTM 2000.The expression and activity of CYP2A13 were detected by real-time quantitative PCR(qRT-PCR), Western blot and Aflatoxin B1(AFB1)/4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone(NNK)cytotoxicity assay and the metabolic activity was compared between CYP2A13-CHO and CYP2A13-POR-CHO.Results Compared with non-transfected cells, the mRNA and protein expression of CYP2A13 in CYP2A13-CHO and CYP2A13-POR-CHO cells both increased significantly.Besides, compared with CYP2A13-POR-CHO, CYP2A13-CHO cells were more sensitive to AFB1 and NNK.Conclusions The Flp-In CHO cell line stably expressing CYP2A13 and with better metabolic activity has been established successfully, which provides a tool for screening of pre-carcinogens that can be metabolically activated by CYP2A13.
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ObjectiveTo explore the mechanism of Shenxiong glucose injection （SGI） in inhibiting hydrogen peroxide （H2O2）-induced oxidative damage in H9c2 cells by tandem mass tags （TMT）-labeled quantitative proteomics. MethodH9c2 cells cultured in vitro were exposed to H2O2 for inducing oxidative damage. The cell viability was measured by cell proliferation and cytotoxicity assay （MTS）， followed by peptide fractionation by high performance liquid chromatography （HPLC） and protein expression detection in H9c2 cells by ultrahigh performance liquid chromatography-mass spectrometry. MaxQuant （v1.5.2.8） was utilized for data retrieval， and the high-resolution mass spectrometry was conducted to screen out differentially expressed proteins， which were then subjected to gene ontology （GO） and Kyoto encyclopedia of genes and genomes （KEGG） enrichment analysis. The protein expression levels of perilipin 2 （Plin2） and tropomyosin 1 （Tpm1） in cells were measured by Western blot. ResultThe spectral analysis yielded 48 608 specific peptide fragments and 5 903 quantifiable proteins. Compared with the model group，the SGI group exhibited 82 differentially expressed proteins，of which 22 were up-regulated and 60 were down-regulated. GO analysis results showed that the differentially expressed proteins were mainly involved in biological processes such as programmed cell death regulation，regulation of cell proliferation，cardiovascular system development， and cell migration. As revealed by KEGG analysis， these proteins were mainly related to peroxisome proliferator-activated receptor （PPAR），focal adhesion，phosphatidylinositol 3-kinase/protein kinase B（PI3K/Akt），and Ras-related protein 1 （Rap1） pathways. Western blot results demonstrated that compared with the model group，SGI significantly increased the Plin2 protein expression and decreased the Tpm1 protein expression （P<0.01），consistent with the proteomics results. ConclusionSGI may inhibit cell apoptosis and antagonize H2O2-induced cell oxidative damage by regulating PPAR，focal adhesion，PI3K/Akt and Rap1 pathways，which should be further verified by subsequent experiments.
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The aim of this paper was to explore the mechanism of Shenxiong Glucose Injection antagonizing apoptosis of H9 c2 cells induced by H_2O_2. H9 c2 cells were pretreated with 1. 7%,3. 4% and 6. 8% Shenxiong Glucose Injection,and then H_2O_2 was introduced to induce apoptosis in vitro. Cell viability was detected by MTS assay,morphological changes of apoptosis were observed by AO/EB fluorescence staining,apoptosis rate was detected by Annexin/PI method,cell expression profile was detected by gene chip technology,the mRNA of PIK3 CA,Bcl-2,Bax,caspase-3 and GAPDH were detected by qRT-PCR,the protein expression levels of PIK3 CA,AKT,P-AKT,Bcl-2,Bax and caspase-3 were detected by Western blot,and the contents of LDH and MDA were detected by kit. The results showed that Shenxiong Glucose Injection of different concentrations significantly increased the viability of H9 c2 cells treated with H_2O_2( P<0. 01),and reversed H_2O_2-induced apoptosis( P< 0. 01). The microarray experiments showed that 138 genes were altered in H9 c2 cells after treatment with Shenxiong Glucose Injection. The differential expression fold of PIK3 CA associated with PI3 K/AKT pathway was 3. 59. The results of qRT-PCR and Western blot showed that Shenxiong Glucose Injection could down-regulate the mRNA and protein expression levels of caspase-3( P<0. 01),up-regulate the mRNA and protein expression level of PIK3 CA and Bcl-2( P<0. 01),and up-regulate the phosphorylation levels of AKT( P<0. 01) in H_2O_2-treated H9 c2 cells. The protective effect of Shenxiong Glucose Injection on H_2O_2 cells injury was significantly inhibited by LY294002,a PI3 K/AKT pathway inhibitor. The results suggested that Shenxiong Glucose Injection may inhibit H_2O_2-induced H9 c2 cells apoptosis by regulating PI3 K/AKT signaling pathway.
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Animals , Rats , Apoptosis , Cell Line , Chromones , Drugs, Chinese Herbal , Pharmacology , Glucose , Morpholines , Phosphatidylinositol 3-Kinases , Metabolism , Proto-Oncogene Proteins c-akt , Metabolism , Signal TransductionABSTRACT
Objective:To discuss the topical action characteristics of the biological transmission of moxibustion heat via temperature collection and numerical modeling.Methods:Temperature of moxibustion was measured at multiple points at a distance of 3 cm to obtain the moxibustion temperature field nephograms by the high-accuracy temperature measure array.Finite element analysis was used to imitate the three-dimensional dynamic distribution of temperature in acupoint tissues.Results:Through numerical analysis,the one-dimensional,two-dimensional and three-dimensional distributions of temperature in human acupoint tissues at 5 min of moxibustion were established.The result showed that moxibustion heat mainly transmitted from the surface of the tissue to the internal,and the influence of moxibustion heat decreased with the depth of the tissue.The analysis of the nephograms of acupoint tissue temperature at 5,10,15 and 20 min of moxibustion showed that with the increase of the moxibustion time,the temperature in acupoint tissues constantly rose,and the transmission depth of moxibustion heat also further expanded inside acupoint.Conclusion:By establishing the three-dimensional dynamic model of heat transmission inside acupoint tissues with the biological parameters of human tissues and the temperature values obtained,this study used finite element analysis software ANSYS 14.0 and discovered the rules in the transmission of heat in body tissues during moxibustion,and the features in moxibustion heat transmission (from the proximal to the distant) and heat penetration (from the surface to the internal).This study provides theoretical and experimental support for the application of moxibustion in clinical practice.
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The cerebral ischemia-reperfusion model was established to investigate the anti-oxidation and anti-apoptosis mechanism of Xinshao formula on the cerebral ischemia reperfusion injury in rats. SD rats were randomly divided into five groups: sham operation group, model group, and low, middle and high-dose Xinshao formula groups (0.31, 0.62, 1.25 g·kg⁻¹). After administration with Xinshao formula for 7 days, the rats were used to establish the cerebral ischemia-reperfusion model. The neurological behavior was evaluated. TTC staining was implemented to determine the volume of cerebral infarction. The levels of ROS, SOD, GSH-PX, NO and iNOS in serum were examined, and the mRNA and protein levels of Bcl-2, Bax and caspase 3 in hippocampal CA1 were detected by qRT-PCR, immunohistochemical assay and Western blot. It is found that Xinshao formula could significantly reduce the symptoms of nervous function and the volume of cerebral infarction in MACO rats. Compared with model group, the rats in Xinshao formula group showed increases in the activities of SOD and GSH-PX (<0.01), and decreases in the activities of iNOS and the contents of NO, ROS and MDA (<0.01). In addition, Xinshao formula could down-regulate the mRNA and protein levels of Bax and caspase 3 (<0.01), and up-regulated those of Bcl-2 (<0.01) in MACO rats. In conclusion, Xinshao formula showed aprotective effect on cerebral ischemia reperfusion injuryin rats, which may be associated with the promotion of anti-oxidation and anti-apoptosis.
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Objective To observe the clinical efficacy of stone needle therapy plus medication in treating early-mid stage senile knee osteoarthritis (KOA).Method Seventy elderly patients with early-mid stage KOA were randomized into a treatment group and a control group, 35 cases each. The control group was intervened by oral administration of Meloxicam tablets, while the treatment group was intervened by stone needle therapy based on the medication. The two groups were treated 5 times a week, for successive 3 weeks. Visual Analogue Scale (VAS) and Lysholm Knee Scoring (LKS) were observed before and after the treatment, and the clinical efficacies of the two groups were also compared. Result The total effective rate was 94.3％ in the treatment group versus 80.0％ in the control group, and the between-group difference was statistically significant (P<0.05). After the treatment, the VAS and LKS scores were significantly changed in both groups (P<0.05). The VAS and LKS scores of the treatment group were significantly different from those of the control group (P<0.05).Conclusion Stone needle therapy plus medication is an effective method in treating early-mid stage senile KOA.
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Objective: To explore the points selection pattern of acupuncture for sleep apnea syndromes by data mining technique. Methods: Clinical literature about acupuncture therapy for sleep apnea syndromes was derived from China National Knowledge Infrastructure (CNKI), Wanfang Academic Journal Full-text Database (Wanfang), Chongqing VIP Database (CQVIP), PubMed and Science Direct between the time that databases were created and March 25th,2017. Relevant excel database was established and descriptive studies and association rules were analyzed. Results: The most frequently used point was Lianquan (CV 23) and the most frequently used meridian was the Stomach Meridian. The analysis of association rules showed that the clinical choice of acupuncture points was highly correlated, among which the combination of the highest degree of confidence and the highest degree of support was Shenmen (HT 7) and Sishencong (EX-HN 1); Lieque (LU 7), lianquan (CV 23) and Zhaohai (KI 6). Conclusion: Acupuncture treatment of sleep apnea syndromes has specific selection rules of points, providing certain references for clinical and scientific research.
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Objective: To observe the effects of different doses of ginger-partitioned moxibustion on serum trefoil factor 1 (TFF1) and mucin 5AC (MUC5AC) levels, as well as the expression of epidermal growth factor receptor (EGFR) in gastric mucosa of rats with spleen deficiency syndrome, therefore, to explore the possible mechanism and the dose-effect characteristics of ginger-partitioned moxibustion in spleen deficiency syndrome. Methods: Seventy-five SPF grade Sprague-Dawley (SD) rats were randomly divided into a blank control group (group A), a model group (group B), a 3 moxa-cone ginger-partitioned moxibustion group (group C1), a 6 moxa-cone ginger-partitioned moxibustion group (group C2) and a 9 moxa-cone ginger-partitioned moxibustion group (group C3) using random number table method, 15 rats in each group. Except group A, rats in the other groups received intragastric administration of 4 ℃200% concentrated Da Huang (Radix et Rhizoma Rhei) to prepare spleen deficiency syndrome model. After successful modeling, rats in group B received no treatment; rats in group C1, C2 and C3 were treated with 3, 6 and 9 moxa-cone ginger-partitioned moxibustion at Zusanli (ST 36) and Zhongwan (CV 12) respectively for 8 continuous days. The general symptom score of rats was observed. The serum levels of TFF1 and MUC5AC were detected by enzyme-linked immunosorbent assay (ELISA). The expression of EGFR protein in gastric mucosa was detected by immunohistochemistry. Results: After the treatment, compared with group A, the spleen deficiency symptom score was increased in group B, the levels of serum TFF1 and MUC5AC, the EGFR protein expression in gastric tissues of group C1, C2 and C3 were significantly increased (all P<0.01); compared with group B, the spleen deficiency scores were decreased in group C1, C2 and C3, and the serum levels of TFF1 and MUC5AC, as well as EGFR protein expression in gastric tissues were increased (all P<0.01). Compared with group C1, the spleen deficiency scores were decreased in group C2 and C3, the serum levels of TFF1 and MUC5AC, and the expression of EGFR protein in gastric tissues were increased (all P<0.01), however, there was no significant difference between group C2 and C3 (all P>0.05). The mechanism may be related to the increase of serum TFF1 and MUC5AC levels and activation of EGFR protein. Conclusion: Ginger-partitioned moxibustion can improve the symptoms, as well as promote the proliferation and repair of gastric mucosa in rats with spleen deficiency. The therapeutic efficacy of 6 or 9 moxa-cone ginger-partitioned moxibustion is better than that of 3 moxa-cone ginger-partitioned moxibustion, while the efficacies are equivalent between 6 and 9 moxa-cone ginger-partitioned moxibustion groups.
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To investigate the protective effects and mechanism of Polygonum orientale flower extract on H₂O₂-induced oxidative damage of human umbilical vein endothelial cells (HUVEC), H₂O₂ was used to induce the oxidativestress damage on HUVEC cells and efforts were made to screen the low, medium and high drug concentrations of P.orientale flower extract. Cell viability was detected by the MTS assay. The content of lactate dehydrogenase (LDH) and malondialdehyde (MDA), and the activities of superoxidedimutase (SOD) and catalase (CAT) were detected by biochemical kits. The mRNA and protein levels of Bax, Bcl-2 were detected respectively by quantitative real time polymerase chain reaction (qRT-PCR) and Western blot. The protein level of cleaved caspase-3 was detected by Western blot. According to the results, the viability of HUVEC cells was reduced to around 55% after being treated with 120 μmol·L⁻¹ H₂O₂ for 0.5 h. Treatment of H₂O₂ also could increase LDH leakage rate and MDA content and attenuate the activities of SOD and CAT, up-regulate the expression level of Bax and cleaved caspase-3, and down-regulate the expression level of Bcl-2. As compared with H₂O₂ model group, P.orientale flower extract of 50-200 mg·L⁻¹ could increase the viability of HUVEC cells, reduce LDH release and MDA content, enhance the activities of SOD and CAT, down-regulate pro-apoptotic protein cleaved caspase-3 and Bax, and up-regulate apoptosis inhibitory protein Bcl-2. In summary, P.orientale flower extract showed a protective effect on H₂O₂-induced HUVEC cells injury, which may result from enhancing the cell capability of clearing the oxygen free radial, decreasing the production of lipid peroxidation and inhibiting apoptosis.
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Objective To investigate the clinical efficacy of gentle moxibustion plus oral Gushen Dingchuan pills in treating the clinical remission stage of bronchial asthma.Methods Ninety patients with bronchial asthma in the clinical remission stage were randomized to groups A,B and C,30 cases each.group A received moxa-stick gentle moxibustion;group B,oral administration of Gushen Dingchuan pills;group C,moxa-stick gentle moxibustion plus oral administration of Gushen Dingchuan pills.The course of treatment was three months in all the three groups.The TCM symptom score and the Asthma Control Test (ACT) score were recorded and serum immunoglobulin E (IgE) content was measured in the two groups before and after treatment.The clinical therapeutic effects were compared between the groups.Results The total efficacy rate was 76.7％ in group A,80.0％ in group B and 93.3％ in group C.there was a statistically significant difference in the total efficacy rate between group C and group A or B (P＜0.05).There were statistically significant pre-/post-treatment differences in the TCM symptom score,the ACT score and serum IgE content in all the three groups (P＜0.01).There were statistically significant post-treatment differences in the TCM symptom score,the ACT score and serum IgE content between between group C and group A or B (P＜0.01,P＜0.05).Conclusion Gentle moxibustion plus oral Gushen Dingchuan pills can improve immunologic function in patients with bronchial asthma in the clinical remission stage.