ObjectiveTo investigate the protective effect of genistein （Gen） on etoposide-induced chondrocyte senescence and its underlying mechanism. MethodsThe C28/I2 cell line was treated with different concentrations of Gen and etoposide， and the cell viability was detected by the CCK-8 assay. The senescence model of C28/I2 chondrocytes was induced by etoposide， with Gen intervention. Senescence-associated β-galactosidase （SA-β-gal） staining was performed to detect the senescence-positive rate and staining characteristics of chondrocytes. The expressions of peroxiredoxin 6 （Prdx6）， cyclin-dependent kinaseto clarify the functional necessity of Prdx6. ResultsCompared with the etoposide group， the C28/I2 chondrocyte viability significantly increased （P<0.01）， the expression ofsenescence-associated proteins p21 and p16 decreased （P<0.01， P<0.05）， the expression of senescence-associated genes p21 and p16 reduced （both P<0.01）， the fluorescence intensity of senescence-associated proteins p21 and p16 was diminished （P<0.05， P<0.01）， and the proportion of SA-β-gal-positive cells decreased （P<0.01） in the Gen+etoposide group. Compared with the Control group， the expression of Prdx6 was downregulated in the etoposide group （P<0.05）. Compared with the etoposide group， the expression of Prdx6 was upregulated in the Gen+etoposide group （P<0.01）. Compared with the Control group， the GPx activity significantly decreased in the si-Prdx6 group （P<0.01）. Furthermore， compared with the si-Prdx6 group， the GPx activity increased in the si-Prdx6+Gen group （P<0.05）. Molecular docking results revealed that Gen formed hydrogen bond interactions with the active site of Prdx6. After Prdx6 knockdown， the expression of senescence-associated genes p21 and p16 and the fluorescence intensity of senescence-associated proteins p21 and p16 both increased in the Gen+etoposide+si-Prdx6 group （both P<0.01）. ConclusionGen can inhibit etoposide-induced senescence of C28/I2 chondrocytes by upregulating the expression of Prdx6. This study provides potential drug targets and experimental basis for the prevention and treatment of chondrocyte senescence-related diseases.

AIM:To appraise the bioequivalence and safety of the test preparation of ritonavir tab-lets and the reference preparation(trade name:Norvir?)in healthy adult subjects under fasting and postprandial conditions.METHODS:This study was a randomized,open-label,single-dose,four-period,fully repeated crossover design bioequivalence study protocol.Thirty-six healthy male and female volunteers were enrolled in the fasting and post-prandial conditions,and a single dose of the test preparation and reference preparation was orally administered.We used liquid chromatography-tan-dem mass spectrometry(LC-MS/MS)to finish the bioassay of the drug concentration of ritonavir in plasma.Pharmacokinetic parameters were statisti-cally analyzed using PhoenixWinNonlin8.1 software(Pharsight,USA)and a non-compartmental model.RESULTS:Under fasting conditions,the pharmacoki-netic parameters of the test and reference prepara-tions:Cmax(792.010±369.282)ng/mL and(856.939±394.427)ng/mL,AUC0-t(6 463.043±2 876.849)ng·mL-1·h and(6 907.690±3 046.132)ng·mL-1·h,AUC0-∞(6 603.617±2 916.352)ng·mL-1·h and(7 051.614±3 093.047)ng·mL-1·h.Here are the pharmacokinetic parameters for both the test prep-aration and the reference preparation in the post-prandial condition:Cmax(574.380±289.566)ng/mL and(615.796±297.382)ng/mL,AUC0-t(5 084.796±2 435.557)ng·mL-1·h and(5 414.167±2 416.952)ng·mL-1·h,AUC0-∞(5 219.144±2 487.793)ng·mL-1·h and(5 551.060±2 490.604)ng·mL-1·h.The 90%confidence interval of the geometric mean ratio of AUC0-t,AUC0-∞,and Cmax for the test preparation and reference preparation lied in the equivalent range of statistics.CONCLUSION:The tested preparation was bioequivalent to the reference preparation un-der fasting and postprandial conditions.

AIM:To appraise the bioequivalence and safety of the test preparation of ritonavir tab-lets and the reference preparation(trade name:Norvir?)in healthy adult subjects under fasting and postprandial conditions.METHODS:This study was a randomized,open-label,single-dose,four-period,fully repeated crossover design bioequivalence study protocol.Thirty-six healthy male and female volunteers were enrolled in the fasting and post-prandial conditions,and a single dose of the test preparation and reference preparation was orally administered.We used liquid chromatography-tan-dem mass spectrometry(LC-MS/MS)to finish the bioassay of the drug concentration of ritonavir in plasma.Pharmacokinetic parameters were statisti-cally analyzed using PhoenixWinNonlin8.1 software(Pharsight,USA)and a non-compartmental model.RESULTS:Under fasting conditions,the pharmacoki-netic parameters of the test and reference prepara-tions:Cmax(792.010±369.282)ng/mL and(856.939±394.427)ng/mL,AUC0-t(6 463.043±2 876.849)ng·mL-1·h and(6 907.690±3 046.132)ng·mL-1·h,AUC0-∞(6 603.617±2 916.352)ng·mL-1·h and(7 051.614±3 093.047)ng·mL-1·h.Here are the pharmacokinetic parameters for both the test prep-aration and the reference preparation in the post-prandial condition:Cmax(574.380±289.566)ng/mL and(615.796±297.382)ng/mL,AUC0-t(5 084.796±2 435.557)ng·mL-1·h and(5 414.167±2 416.952)ng·mL-1·h,AUC0-∞(5 219.144±2 487.793)ng·mL-1·h and(5 551.060±2 490.604)ng·mL-1·h.The 90%confidence interval of the geometric mean ratio of AUC0-t,AUC0-∞,and Cmax for the test preparation and reference preparation lied in the equivalent range of statistics.CONCLUSION:The tested preparation was bioequivalent to the reference preparation un-der fasting and postprandial conditions.

Objective : To explore the effects and mechanisms of Kobophenol A ( KPA) on lipopolysaccharide ( LPS) -induced M1 macrophage polarization，and to provide a theoretical basis for the treatment of inflammatory immune diseases and the development of new drugs． Methods: The M1 macrophage polarization model of RAW264. 7 was established by LPS induction，and the peroxiredoxin 6 ( Prdx6) knockdown model was constructed using the Prdx6 inhibitor MJ33 and Prdx6-siRNA．RAW264. 7 cells，a mouse macrophage cell line，were treated with various concentrations of KPA． Cell viability was assessed using the CCK-8 assay．The expression levels of Prdx6 and M1 macrophage polarization-related proteins，including inducible nitric oxide synthase ( iNOS) and cyclooxygenase-2 ( COX-2) ，were detected by Western blot and immunofluorescence staining．The expression levels of Prdx6 and M1 macrophage polarization-related genes iNOS，interleukin-6 ( IL-6) ，and tumor necrosis factor α ( TNF-α) ，were measured by RT-qPCR. Flow cytometry was employed to detect the expression of cluster of differentiation 86 ( CD86) ，a marker of M1 macrophages． Results: Compared with the LPS-induced M1 macrophage polarization model ， KPA significantly reversed the morphological changes of M1 macrophage polarization in RAW264. 7 macrophages and decreased the expression of M1 macrophage polarization-related proteins iNOS，COX- 2，CD86 and related genes iNOS，IL-6，TNF-α ( all P＜0. 05) ．In addition，LPS significantly downregulated the expression of Prdx6 in RAW264. 7 macrophages，while KPA upregulated the expression of Prdx6．Moreover，treatment with the Prdx6 inhibitor MJ33 significantly upregulated the expression of iNOS，a marker of M1 macrophage polarization，in RAW264. 7 macrophages，whereas treatment with KPA significantly downregulated the expression of iNOS ( all P＜0. 05) ． Conclusion KPA inhibits LPS-induced M1 polarization of RAW264. 7 macrophages by upregulating the expression of Prdx6．

Objective To investigate the role of Taraxasterol(TAR)on ferroptosis in chondrocytes induced by Erastin.Methods The C28/I2 chondrocyte line was treated with Erastin to construct the ferroptosis model of chon-drocytes in vitro and the experiments were divided into Control,Erastin,TAR,and TAR+Erastin groups.Cell via-bility was detected by the CCK-8 assay.Cytotoxicity was detected by the lactate dehydrogenase(LDH)kit and the Calcein/PI cytokinesis kit.Flow cytometry was used to detect lipid reactive oxygen species(ROS).The intracellular glutathione(GSH)content was detected by GSH kit.Mitochondrial membrane potential was detected by JC-1 stai-ning and RH123 staining.ACSL4 and GPX4 protein expression and the key indicators of ferroptosis were detected by Western blot.Results TAR restored the decreased cell viability of C28/I2 chondrocytes induced by Erastin treatment as well as reduced Erastin-induced cytotoxicity(P＜0.01).Compared with the control group,the level of intracellular lipid ROS increased(P＜0.01)and the content of GSH decreased(P＜0.01)after treatment with Erastin,while TAR could reduce the production of lipid ROS(P＜0.01)and increase the content of GSH(P＜0.01).TAR restored mitochondrial membrane potential in C28/I2 chondrocytes ferroptosis,decreased ACSL4 pro-tein expression(P＜0.01)and increased GPX4 protein expression(P＜0.01).In addition,TAR restored the re-duced cell viability caused by IL-1 β treatment.Conclusion TAR can inhibit Erastin induced ferroptosis in C28/I2 chondrocytes,which may be related to the regulation of ACSL4 and GPX4 protein expression.

Objective To explore the effect of 2-aminoethoxy-diphenyl borate(2-APB)on H2O2-induced chondro-cyte apoptosis and its mechanism.Methods The experiment was divided into control group,H2O2 group,2-APB group and H2O2+2-APB group.CCK-8 method was used to detect the cell viability of each group;The effect of 2-APB on the morphological changes of chondrocytes induced by H2O2 was observed under microscopy;TUNEL meth-od and flow cytometry were used to detect chondrocyte apoptosis;Flow cytometry was used to detect Lipid reactive oxygen species(ROS);Western blot was used to detect the protein expressions of Cleaved-PARP,p-PKCα and HIF-1α in H2O2-induced cells by 2-APB;Immunofluorescence was used to detect the fluorescent expression of HIF-1α in cells induced by H2O2 by PKCα inhibitor BIM-1.Results 2-APB inhibited H2O2-induced apoptosis in chon-drocytes,and the inhibitory effect was the most significant when the concentration of 2-APB was 100 pmol/L(F=235.80,P＜0.01);22-APB could inhibit the positive rate of H2O2-induced apoptosis of chondrocytes(F=114.80,P＜0.01)and the level of ROS(F=52.99,P＜0.01).and inhibited the expression of Cleaved-PARP(F=10.10,P＜0.05),p-PKCα(F=24.56,P＜0.05)and HIF-1α proteins(F=6.85,P＜0.05).The PKCα in-hibitor BIM-Ⅰ could inhibit the increase in HIF-1α fluorescence intensity caused by H2O2.Conclusion 2-APB can inhibit chondrocytes apoptosis induced by H2O2 through the PKCα/HIF-1α pathway and thus protect chondro-cytes.

AIM: To compare the pharmacokinetic behavior of a single oral sorafenib tosylate tablets in Chinese healthy subjects under fasting conditions and evaluate the bioequivalence of the test reagent (T) and the reference reagent (R). METHODS: A single-center, randomized, open-labeled, two-agent, three-period, three-sequence (TRR, RTR, RRT), and partially repeated crossover trial design was adopted. The trial was administered once per cycle (0.2 g) under fasting conditions. 36 healthy subjects were randomly divided into 3 groups, each with 12 cases. RESULTS: Thirty-six healthy subjects (9 females, average age 31 years) were enrolled in the trial. The upper limits of the one-sided 95% confidence interval of the pharmacokinetic parameters C

Objective    To discuss the safety, feasibility and short-term clinical efficacy of thoracoscopic anterior mediastinal mass resection in lithotomy position via subxiphoid approach or lateral position via transthoracic approach. Methods    A total of 44 patients suffering anterior mediastinal tumor enrolled, including 21 patients (10 males and 11 females as a trial group) with an average age of 43.6±11.8 years who have been performed thoracoscopic anterior mediastinal tumor resection in lithotomy position via subxiphoid approach and 23 patients (13 males and 10 females as a control group) with an average age of 45.3±10.8 years who have been performed thoracoscopic anterior mediastinal tumor resection in lateral position via transthoracic approach. The clinical efficacy of the two groups was compared. Results    Postoperative chest drainage time (3.8±1.3 d vs. 5.0±1.8 d, P=0.017), postoperative drainage volume (238.8±66.2 mL vs. 467.2±120.0 mL, P=0.000), postoperative mean  visual analogue score at 24 h (2.5±0.9 point vs. 4.9±1.0 point, P=0.000), times of self-pressure analgesic pump (3.7±0.9 vs. 8.4±2.0, P=0.000), duration of postoperative hospital stay (4.7±1.3 d vs. 7.4±3.1 d, P=0.000) and hospitalization cost (34±8 kyaun vs. 44±11 kyuan P=0.001) in the trial group were all better than those in the control group. There was no significant difference between the two groups in surgical duration (59.0±18.1 min vs. 60.4±16.4 min) (P>0.05). During follow-up, no recurrence or metastasis occurred in either group. Conclusion    Compared with the lateral position through the transthoracic approach, the lithotomy position through subxiphoid approach of thoracoscopic anterior mediastinal mass resection is safe and feasible, and has certain advantages.

Objective To investigate the effect of surgical treatment of necrotizing enterocolitis (NEC) with different surgical ages.Methods From January 2014 to December 2017,105 neonates with NEC in our hospital were divided into early operation group (operation age ＜ 7 days,n =47) and late operation group (7 ＜ operation age ＜ 28 days,n =58).The general data,surgical indications,intraoperative conditions,surgical methods,postoperative complications,and postoperative survival rates were compared between the two groups.Results Among the 105 neonates with NEC,74 were male and 31 were female.The average birth weight was (2 398 ± 927)g,and the average gestational age was (35 ± 4)weeks.Compared with the early operation group,the late operation group had lighter birth weight,smaller gestational age and higher rate of respiratory failure (P ＜ 0.05).There was no significant difference between the two groups in the proportion of surgical indications (diffuse peritonitis,pneumoperitoneum,and medical treatment ineffective) (P ＞ 0.05).The necrosis rate of small intestine in the late operation group was higher than that in the early operation group,but the necrosis rate of small intestine and colon was lower than that in the early operation group (P ＜ 0.05).There was no significant difference in the proportions of the two groups in the surgical methods (enterostomy,intestinal resection and anastomosis and enterostom,exploratory laparotomy,abdominal drainage,and intestinal resection and anastomosis) (P ＞ 0.05).The incidence of intestinal stenosis in early operation group was higher than that in late operation group (P ＜ 0.05).The survival rate of early operation group was 78.7％,while that of late operation group was 63.8％,with no significant difference (P ＞ 0.05).Conclusions The patients with NEC who were operated within 1 week after birth are more common in term infants and with colon necrosis,and are more likely to occur intestinal stenosis after surgery.The patients with NEC who were operated after 1 week of birth are more common in prematures and low-birth-weight patients,and are often associated with respiratory failure.Pneumoperitoneum and diffuse peritonitis are common surgical indications for NEC.Enterostomy is the major surgical method.Choosing the right timing and surgical method can improve the prognosis of patients with NEC.

Objective To summarize the pathology of congenital intestinal atresia,the incidence and prenatal diagnosis rate of different types,and to analyze the location and type of intestinal atresia as well as the factors that affect the mortality of various types of intestinal atresia.Method We retrospectively analyzed the clinical data of 147 children with congenital intestinal atresia from January 2013 to March 2016,including gender,gestational age,parity,prenatal diagnosis or not,delivery methods,hospital admission,surgical methods,findings during surgery,combined malformations,complications and prognosis.They were analyzed statistically.Result A total of 147 cases,including 69 males and 78 females were enrolled.There were 40 premature infants and 107 full term cases.Twins were found in 3 cases.Hospital admission age range from 1 hour to 62 days;admission weight range from 1 480 g to 4 200 g;32 cases were diagnosed before birth.2 cases were abandoned before surgery because of trisomy 21.Postoperatively,the occlusion sites was confirmed as following:67 cases (46.2％) in ileum,43 cases (29.7％) in jejunum,26 cases (17.9％) in duodenum,and 9 cases (6.2％) in colon.The pathological types were as following:type Ⅰ 42 cases (29.0％),type Ⅱ 8 cases (5.5％),type Ⅲa 65 cases (44.8％),type Ⅲb 15 cases (10.3％) and type Ⅳ 15 cases (10.3％).22 cases (14.9％) were died because of refusal of treatment:7 cases were due to short bowel syndrome and meconium peritonitis,6 cases were due to postoperative chronic pseudo-obstruction,and 5 cases had anastomotic leakage requiring reoperation.1 case had postoperative enterocolitis and gave up treatment,1 case had anastomotic leak and sever systemic post-surgery infection and gave up further treatment,and 2 cases gave up because of 21-trisomy syndrome.Conclusion The operation plan of intestinal atresia should be based on the location and type of the blockade;the location and complications of the blockade (pseudo-obstruction,short bowel syndrome,and anastomotic leakage) are important factors affecting the treatment and prognosis.