OBJECTIVE:To study the time-effect and dose-effect of paeonol on the apoptosis of knee osteoarthritis(OA)artic-ular chondrocyte in rabbits and the mRNA expression of its related protein Bcl-2 and Bax. METHODS:60 big-ear rabbits were ran-domly divided into normal (normal saline) group,model (normal saline) group,paeonol high-dose,medium-dose and low-dose groups and triamcinolone acetonide(positive drug)group,with 10 rabbits in each group. Except for normal group,OA model was induced by right knee anterior cruciate ligament (ACLT) and the medial meniscus 1/3 resection in those groups. After modeling, different doses of paeonol(0.8,0.4,0.2 mg/kg),triamcinolone acetonide 0.2 mg/kg were injected into right articular cavity twice a week. 4 weeks and 8 weeks after administration,articular cartilage specimens were collected. Ultrapathological structure changes of articular chondrocytes were observed by electron microscope. Apoptosis of cartilage cell was observed by TUNEL and apoptotic index was calculated. mRNA expression of apoptosis related genes of Bcl-2 and Bax in articular cartilage tissue of rabbits were de-tected by in situ hybridization technique. RESULTS:Compared with normal group,articular chondrocyte of model group showed early and middle stage apoptosis morphology change after 4 and 8 weeks,and apoptosis index increased significantly and the mRNA expression of Bcl-2 and Bax was up-regulated (P<0.01);4 and 8 weeks later after administration,compared with model group,apoptosis index decreased and mRNA expression of Bax was down-regulated in paeonol groups,while mRNA expression of Bcl-2 was up-regulated(P<0.05 or P<0.01). Electron microscopy ultrastructural observation showed articular chondrocyte of pae-onol high-dose and middle-dose groups were in early stage of apoptosis.CONCLUSIONS:Paeonol can slow down articular chondro-cyte degeneration and destroy in OA model rabbits in time and dose dependently. Its mechanism may be associated with expression up-regulation of Bcl-2 and expression down-regulation of Bax.

Objective To develop a method for determination of iridoid glycosides in Morinda officinalis How.and optimize the extraction methods for iridoid glycosides in Morinda officinalis How.Methods The iridoid glycosides, including monotropein, deacetyl asperulosidic acid,asperulosidic acid and asperuloside as standards, HPLC method was developed to determine the content of iridoid glycosides in Morinda officinalis How.The separation was performed on Venusil MP C18 (250 mm×4.6 mm, 5 μm) column.The mobile phase was acetonitrile (A)-0.2% phosphoric acid and 0.01 disodium hydrogen phosphate buffer salt (B) with gradient elution (0-12 min, 1%-2% A;12-30 min, 2%-25% A).The detection wavelength was 235 nm.The flow rate was set at 1.0 ml/min and the column temperature at 25 ℃.The injection volume was 20 μl.Single factor analysis and orthogonal test were used to optimize extraction method of iridoid glycosides in Morinda officinalis How.Results Monotropein, deacetyl asperulosidic acid, asperulosidic acid and asperuloside showed good linearity (r>0.999 5) in the ranges of 0.375-12 μg, 0.13-4.16 μg, 0.016-0.516 μg and 0.012-0.384 μg, respectively.This validated method has good repeatability, precision, recovery and stability.It was conformed to meet the requirements and regulation.The optimal extraction method included soaking the raw materials with 16 times of 10% ethanol for 9 h, and then extraction by percolation with the flow rate of 0.8 BV/h.Conclusion The HPLC method sensitively and precisely determined the content of iridoid glycosides in Morinda officinalis How.The optimized extraction method extracted these constituents effectively.

Objective: To understand the mortality and influencing factors on injecting drug users (IDUs) with HIV/AIDS, in Guizhou province, 1996-2015. Methods: A retrospective cohort study was conducted on IDUs with HIV/AIDS that were reported through national comprehensive HIV/AIDS information system, in Guizhou province during 1996-2015. Cox proportional hazard regression model was used to analyze the influencing factors on the mortality of HIV/AIDS. Results: A total of 3 958 cases of IDUs with HIV/AIDS were recruited in this study, with all-cause mortality rate of 44.01% (1 742/3 958) and total mortality rate of 7.80/100 person-years, respectively. The median survival time between diagnosis and death was 8.08 years. Mortality rate was 3.57/100 person-years in the group receiving antiretroviral therapy (ART). The mortality appeared to be 4.08/100 person-years in the group who were on methadone maintenance treatment (MMT). Data from the multiple regression analysis indicated that factors of gender, ethnicity, age when HIV/AIDS diagnosis was made, CD₄⁺T lymphocyte (CD₄) count at the first testing, ART and MMT were significantly associated with deaths among these people. The risk of death in females was 0.82 times (95%CI: 0.69-0.98) higher than that in males. The risk of deaths among the ethnic minority subjects was 1.39 times (95%CI: 1.21-1.60) higher than that of the Hans. The risk of death appeared to be 2.44 times higher (95%CI: 1.07-5.56) in the over-50-year of age group than in the <20 year-old group, when HIV/AIDS was diagnosed for the first time. The risk of death in CD₄ ≥500/μl group in the first time was 0.27 times (95%CI: 0.22-0.32) more than CD₄ <200/μl group in the firs time. The risk of death in cases who were treated with ART or MMT was 2.83 times (95%CI: 2.45-3.26) and 1.35 times (95%CI: 1.15-1.59) higher than those who did not receive any treatment, respectively. Conclusion Higher risks on death seemed to be related to the following factors: being male, older age at the time of diagnosis, lower CD₄ at diagnosis, not on ART or MMT among the IDUs with HIV/AIDS in Guizhou province, between 1996-2015.

Cancer is the leading cause of death in children. With the development of family participatory care, it has become a trend to use home chemotherapy for children with cancer in stable stages. Therefore, this article introduces the current situation of home chemotherapy medication safety, elaborates on the influencing factors of family chemotherapy medication safety for cancer children from two aspects including oral chemotherapy and intravenous chemotherapy, and proposes intervention measures to optimize the family chemotherapy medication process for cancer children, improve their home chemotherapy medication ability and effectively ensure the medication safety of cancer children.

Objective:To explore the expression features of cytochrome C oxidase subunit Ⅰ (MT-CO1), BCL2 interacting protein 3 (BNIP3) and interleukin (IL)-1β in the liver of MRL/lpr lupus mice.Methods:The mRNA and protein levels of MT-CO1, BNIP3, IL-1β, p16 and p21 in lupus mice and control mice were detected by polymerase chain reaction (PCR) and Western blot, the IL-1β expression site were detected by hematoxylin and eosin (HE) staining and immunohistochemical method, and themalondialdehyde (MDA) was detected by colorimetry. Hepatocytes and macrophages were stimulated with lipopolysaccharide (LPS), while hepatocytes were also cultured with supernatants obtained after macrophages stimulated with LPS, and the mRNA and protein levels of MT-CO1, BNIP3 and LC3B, as well as p16 and p21 expression, were determined by qPCR and Western blot. The expression of mitochondrial reactive oxygen species (mtROS) was detected by immunofluorescence. One way Analysis of Variance (ANOVA) was used to compare the mean of each group, and LSD method was used to compare the means of multiple samples, and Tamhane's T2 method was used to compare the means of multiple samples when the variance was uniform. Results:The results of PCR showed that the mRNA levels of MT-CO1 and BNIP3 in the liver tissue of the lupus group (0.14±0.04; 0.16±0.05) were significantly lower than those of the control group (0.11±0.04; 0.16±0.06), and the differences were statistically significant ( t=7.16, P<0.001; t=4.54, P<0.001). The expression levels of IL-1β, p16 and p21 in the lupus group (2.06±0.69; 0.37±0.14; 0.16±0.06) were significantly higher than those of the control group (0.23±0.06; 0.25±0.08; 0.11±0.04) ( t=9.58, P<0.001; t=24.35, P<0.001; t=22.36, P<0.001). The results of Western blot were consistent with those of PCR. HE staining showed lymphocyte infiltration in the liver tissue of lupus mice, and immunohistochemistry showed IL-1β in the liver tissue of lupus mice. The positive cells were mainly concentrated in the sinusoids, and the expression of hepatic parenchymal cells was not rearkable. The content of MDA in liver tissue of the lupus group (0.19±0.10) was higher than that of the control group (0.17±0.09), and the difference was statistically significant ( t=4.33, P=0.005). LPS directly stimulated AML12 hepatocytes (0.069±0.028; 0.17±0.07). The PCR results showed that compared with the control group (0.176±0.072; 0.08±0.03), the expression of MT-CO1, and BNIP3 were not significantly different ( t=1.01, P=0.337; t=0.88, P=0.399). The expression of IL-1β was significantly higher when incubated with the supernatants of LPS stimulated macrophages (0.28±0.09) compared than that of the control group (0.15±0.05) ( t=28.26, P<0.001). The results of PCR showed that the mRNA levels of MT-CO1 and BNIP3 in the LPS stimulated group (0.046±0.026; 0.17±0.05) were significantly lower than those in the control group (0.143±0.083; 0.18±0.06), and the differences were statistically significant ( t=7.52, P<0.001; t=4.24, P<0.001), The expression of p16 and p21 in LPS stimulated group (0.29±0.09; 0.27±0.09) were significantly higher than those in the control group (0.18±0.06; 0.22±0.07) ( t=13.54, P<0.001; t=8.69, P<0.001). The results of Western blot were consistent with those of PCR. Immunofluorescence showed that the fluorescence intensity of mtROS in LPS stimulated group (0.25±0.10) was higher than that in the control group (0.08±0.03), and the difference was statistically significant ( t= 4.86, P<0.001). Conclusion:Immune-mediated inflammation in the liver tissue of lupus mice can stimulate liver parenchymal cells to cause intracellular mitochondrial dysfunction. However, the mechanism of liver organ damage in lupus mice is not limited to the immune-mediated inflammation of immune active cells, but also include parenchymal cell mitochondrial dysfunction.

Objective To systematically evaluate the accuracy of computer navigation technique in the positioning of bone tunnel location of anterior cruciate ligament reconstruction(ACLR) and its effect on post-operative functional recovery.Methods The controlled trials of computer navigation-assisted ACLR in the da-tabases of Cochrane Library,PubMed,Embase,CNKI,Wanfang and VIP Database were retrieved.The retriev-al time limit was from the establishment of the database to August 2023.According to the inclusion and exclu-sion criteria,the NoteExpress V3.0 software was used to screen the literatures,the Cochrane risk bias assess-ment tool was used to evaluate the quality of the included literatures,and the RevMan5.4 software was used for conducting the meta analysis.Results A total of 10 trials involving 705 patients were included,including 354 cases in the navigation group and 351 cases in the conventional group.The meta analysis results showed that compared with the conventional group,the location positioning of bone tunnel in the navigation group was significantly improved[in femoral side (MD=5.59,95％CI:1.21-9.97,P=0.01) and tibial side (MD=1.32,95％CI:0.20-2.43,P=0.02).However there were no statistically significant differences in the IKDC scores (MD=1.76,95％CI:-0.17 to 3.70,P=0.07),Lysholm scores (MD=0.49,95％CI:-0.16 to 1.14,P=0.14),Tegner scores (MD=-0.08,95％CI:-0.35 to 0.20,P=0.58),KT-1000 anterior shift dis-tance (MD=0.01,95％CI:-0.49 to 0.52,P=0.96),the positive rate of Lachman test (RD=-0.01,95％CI:-0.09 to 0.07,P=0.75) and the positive rate of axial shift test (RD=-0.09,95％CI:-0.22 to 0.04,P=0.20).Conclusion The computer navigation technology is conducive to elevate the accuracy of tibial and femoral tunnel positioning in ACLR,but could not improve the postoperative functional recovery of the pa-tients.

BACKGROUNDTo evaluate and compare the effects and toxicity of the domestic product of nrhTNF combined with chemotherapy in the trial group and chemotherapy alone in the control group in the treatment of patients with non-small cell lung cancer (NSCLC).

METHODSNinety patients with NSCLC in multicenter were randomly devided into trial group and control group. Each group had 45 patients. Chemotherapy with CAP regimen was given for the patients in the trial group. Meanwhile, nrhTNF injection of 4×10⁶U/m ² was also given from the 1st to 7th days, the 11th to 17th days on the chemotherapy course. Twenty-one days were as a cycle, 2 cycles were given each patients. Chemotherapy alone with CAP regimen was given in the control group. The chemothepeutic effects and toxicity were observed and compared between the two groups after the therapy.

RESULTSOf the 90 patients, 3 cases in each group were out of the trial because of economy. The other 84 cases (each group had 42 patients) could be used to analyze and evaluate the clinical effects and toxicity. The response rate of chemotherapy was 47.62% (20/42) in the trial group and 19.05% (8/42) in the control group (P=0.002) respectively. The KPS was 85.02±10.74 in the trial group, and 81.35±9.63 in the control group (P=0.038). No significant difference of degree III+IV toxicity was observed between the trial group and control group (P > 0.05). The side effects related to nrhTNF included slight fever, cold like symptoms, pain, and red and swelling in injection site. All of them were mild and didn't need any treatment and disappeared after the therapy.

CONCLUSIONSThe results demonstrate that the effects of domestic nrhTNF combined with chemotherapy can remarkably higher than that of chemotherapy alone in the treatment of NSCLC. It is able to increase the sensitivity to chemotherapy and improve the quality of life of the patients. The toxicity is also slight and is worth to expand clinical use, so as to further evaluate its effect and toxicity.

Background Arsenic is a well-known environmental toxicant. Hepatic fibrosis could occur dueto excessive or long-term exposure to arsenic, while associated molecular mechanisms remain undefined. Mitogen-inducible gene 6 (Mig-6) exhibits a protective effect on numerous diseases or cancers. However, the specific role of Mig-6 in the mechanisms of arsenite-induced hepatic fibrosis remains indistinct. Objective To investigate the specific role of Mig-6 in the activation of hepatic stellate cells (HSC) and the deposition of extracellular matrix (ECM) induced by sodium arsenite (NaAsO₂). Methods Human hepatic stellate cells (Lx-2) were treated with 0, 1.875, 3.75, 7.5, and 15 μmol·L⁻¹ of NaAsO₂ for 24 h, or with 7.5 μmol·L⁻¹ NaAsO₂ for 0, 12, 24, 48, and 72 h. Additionally, Lx-2 cells were transfected by pcDNA3.1(+)/Mig-6, then treated with 7.5 μmol·L⁻¹ NaAsO₂ for 24 h; a blank control group, a pcDNA3.1(+)-control group, a pcDNA3.1(+)/Mig-6 group, and an arsenic (7.5 μmol·L⁻¹ NaAsO₂) group were also set up. After transfection, the cells and culture supernatants were collected, and the protein levels of Mig-6, α-smooth muscle actin (α-SMA), and transforming growth factor-β1 (TGF-β1) in Lx-2 cells were identified by Western blotting analysis; moreover, the secretion levels of main ECM components in supernatants such as hyaluronic acid (HA), laminin (LN), collagens IV (COL-IV), and procollagen-III (PIIINP) were tested by ELISA. Results The Mig-6 expression decreased in the 3.75, 7.5, and 15 μmol·L⁻¹ NaAsO₂ groups (0.561±0.095, 0.695±0.048, and 0.401±0.030) compared to the control group (1.000±0.000) in Lx-2 cells (P<0.05). After administration with 7.5 μmol·L⁻¹ of NaAsO₂ for 24, 48, and 72 h, the Mig-6 expression (0.856±0.036, 0.515±0.077, 0.491±0.060) decreased compared with the 0 h group (1.000±0.000) (P<0.05). After over-expression of Mig-6, the results of Lx-2 activation related protein levels showed that compared to the control group, the α-SMA and TGF-β1 expression were up-regulated in the arsenic group (P<0.05); meanwhile, the α-SMA and TGF-β1 in the Mig-6 over-expression combined arsenic exposure group reduced compared to the arsenic (7.5 μmol·L⁻¹) group (P<0.05). The results of ELISA showed that compared with the control group, the HA, LN, PIIINP, COL-IV in the arsenic group were up-regulated (P<0.05); while compared to the arsenic group, the HA, LN, PIIINP, and COL-IV in the Mig-6 over-expression combined with arsenic exposure group were decreased (P<0.05). Conclusion Arsenic down-regulates Mig-6 expression in HSC, and over-expression of Mig-6 can reverse the activation of HSC and ECM deposition induced by arsenic exposure. It suggests that Mig-6 plays a protective role in arsenic-induced HSC activation and ECM deposition.