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 causal association between the onset of gastroesophageal reflux disease (GERD) and migraine and to provide genetic evidence, a two-sample bidirectional Mendelian randomization (MR) method was used in this study. 【Methods】 Single nucleotide polymorphism (SNP) information for both samples was obtained from publicly available genome-wide association study (GWAS) databases, in which the appropriate SNPs were selected as instrumental variables, and then bidirectional MR analysis used five MR analysis methods including inverse variance weighting (IVW), MR-Egger regression, weighted median, weighted mode and simple mode methods, followed by sensitivity analysis. 【Results】 IVW showed positive results of forward MR analysis with GERD as exposure [OR=1.398 7, 95%CI (1.181 7-1.655 6), P=9.59×10^-5] , while no positive significance of reverse MR analysis results with migraine as exposure (P>0.05). The same results were obtained in methods other than MR-Egger method. Meanwhile, none of the instrumental variables were found to be horizontally polytomous (P=0.92, P=0.64), and the results were robust after the leave-one-out method to exclude single SNPs. 【Conclusion】 There may be a unidirectional causal association between GERD and migraine, and GERD is a risk factor for migraine development.

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.

Objective To investigate the liver disease phenotypes and clinical features of patients with different genotypes of Wilson's disease(WD).Methods A retrospective analysis was performed for 163 patients with WD who were diagnosed and underwent genetic testing in The Fifth Medical Center of Chinese PLA General Hospital from August 2008 to June 2023,and clinical manifestations,laboratory examination,pathological examination,imaging examination,and ATP7B genetic testing results were collected.According to ATP7B gene mutation,the patients were divided into groups as follows:R778L mutation group and non-R778L mutation group;P992L mutation group and non-P992L mutation group;truncation mutation group and non-truncation mutation group.Liver disease phenotypes and clinical features were analyzed for the patients with c.2333G>T/p.R778L mutation(R778L mutation),c.2975C>T/p.P992L mutation(P992L mutation),and truncation mutation of the ATP7B gene.The Mann-Whitney U test or the Kruskal-Wallis H test was used for comparison of continuous data between groups,and the chi-square test or the Fisher's exact test was used for comparison of categorical data between groups.Results The 163 patients with WD had varying severities of liver disease phenotypes,among whom 121(74.23%)were diagnosed with chronic liver disease,36(22.09%)were diagnosed with decompensated cirrhosis,and 6(3.68%)were diagnosed with fulminant WD,and in addition,there were 5 patients(2 with chronic liver disease and 3 with decompensated cirrhosis)with neurological abnormalities.For the 163 patients with WD,R778L mutation(with an allele frequency of 28.2%)was the most common mutation in the ATP7B gene,followed by P992L mutation(with an allele frequency of 12.6%),and truncation mutation showed an allele frequency of 11.0%.There was no significant difference in the distribution of the three mutations across different liver disease phenotypes(P>0.05).The R778L mutation group had a significantly lower level of ceruloplasmin(CP)than the non-R778L mutation group[0.04(0.02-0.08)g/L vs 0.08(0.03-0.13)g/L,Z=-2.889,P=0.004].Compared with the non-P992L mutation group,the P992L mutation group had significantly higher levels of alanine aminotransferase[135.0(80.5-237.0)U/L vs 80.5(36.0-173.3)U/L,Z=2.684,P=0.007]and aspartate aminotransferase[121.4(77.0-195.0)U/L vs 84.0(39.0-123.3)U/L,Z=3.388,P<0.001].Compared with the non-truncation mutation group,the truncation mutation group had significantly lower levels of CP[0.03(0.02-0.08)g/L vs 0.06(0.03-0.11)g/L,Z=-3.136,P=0.002]and serum copper[3.20(2.15-5.00)mg/L vs 4.20(2.60-7.50)mg/L,Z=-2.296,P=0.025].Conclusion R778L mutation,P992L mutation and truncation mutation are not associated with liver disease phenotype in WD patients;however,R778L mutation is associated with a lower level of CP,P992L mutation is associated with higher levels of ALT and AST,and truncation mutation is associated with lower levels of CP and serum copper.

As an important part of tumor microenvironment, neutrophils are poorly understood due to their spatiotemporal heterogeneity in tumorigenesis. Here we defined, at single-cell resolution, CD44-CXCR2- neutrophils as tumor-specific neutrophils (tsNeus) in both mouse and human gastric cancer (GC). We uncovered a Hippo regulon in neutrophils with unique YAP signature genes (e.g., ICAM1, CD14, EGR1) distinct from those identified in epithelial and/or cancer cells. Importantly, knockout of YAP/TAZ in neutrophils impaired their differentiation into CD54+ tsNeus and reduced their antitumor activity, leading to accelerated GC progression. Moreover, the relative amounts of CD54+ tsNeus were found to be negatively associated with GC progression and positively associated with patient survival. Interestingly, GC patients receiving neoadjuvant chemotherapy had increased numbers of CD54+ tsNeus. Furthermore, pharmacologically enhancing YAP activity selectively activated neutrophils to suppress refractory GC, with no significant inflammation-related side effects. Thus, our work characterized tumor-specific neutrophils in GC and revealed an essential role of YAP/TAZ-CD54 axis in tsNeus, opening a new possibility to develop neutrophil-based antitumor therapeutics.
Humans ; Animals ; Mice ; Adaptor Proteins, Signal Transducing/metabolism* ; Transcription Factors/metabolism* ; Stomach Neoplasms/pathology* ; Neutrophils/pathology* ; Signal Transduction/genetics* ; YAP-Signaling Proteins ; Tumor Microenvironment ; Hyaluronan Receptors/genetics*

Objective:To analyze the biomechanical characteristics of bilateral lumbo-iliac fixation by finite element analysis after digital optimization of a safe screw pathway.Methods:Firstly, a healthy adult male volunteer was recruited for this study who underwent CT scanning of the lumbus and pelvis, with a slice thickness of 0.625 mm.The original data were imported into Mimics 16.0 software for 3D lumbar and pelvic reconstruction.At the same time, a geometric screw pathway and a free screw pathway were generated in the pedicles of L4 and L5 and ilium.Secondly, all 3D models were imported into Ansys 12 software to assemble a finite element model.The displacements and von Misses stress distribution were measured in the conditions of forward bending, backward extending, lateral bending to the left and right, and left and right rotations in the models of 2 kinds of pathway after gravity axial load of 500 N was applied.Results:The models of 2 different pathways were similar in comprehensive displacement and stress in the conditions of forward bending, lateral bending and backward extending, and in the maximum stress of the screw in the conditions of forward bending and lateral bending.However, the comprehensive displacement and stress in the condition of rotation and the stress of the screw in the conditions of backward extending and rotating in the model of geometric screw pathway were obviously smaller than those in the model of free screw pathway.The maximum pelvic stress was on the screw surface between 190 and 260 MPa, concentrated on the bending area of the 2 screws connecting the sacrum and vertebral body.The lumbar-iliac fixation model showed high com-pressive stiffness.The maximum von Misses stress appeared in the L4-L5 pedicle screw on the longitudinal rod.Under rotational loading, the upper longitudinal rod connecting the L4-L5 pedicle screw and the iliac bone screw bore the maximum stress.Conclusions:Our finite element analysis shows that double verte-bral bodies and long screw fixation are effective ways of iliolumbar fixation.The geometric screw path optimized by digital analysis exhibits better fatigue resistance, rotation resistance and overall stability.Internal lum-bo-iliac fixation can effectively share the stress of vertebral bodies.

The Alataw Pass, near the Ebinur Lake Wetland (northwest of China) and Taldykorgan (east of Kazakhstan), is a natural habitat for wild rodents. To date, little has been done on the surveillance of Bartonella spp. and Wolbachia spp. from fleas in the region. Here we molecularly detected Bartonella spp. and Wolbachia spp. in wild rodent fleas during January and October of 2016 along the Alataw Pass-Kazakhstan border. A total of 1,706 fleas belonging to 10 species were collected from 6 rodent species. Among the 10 flea species, 4 were found to be positive for Wolbachia, and 5 flea species were positive for Bartonella. Molecular analysis indicated that i) B. rochalimae was firstly identified in Xenopsylla gerbilli minax and X. conforms conforms, ii) B. grahamii was firstly identified in X. gerbilli minax, and iii) B. elizabethae was firstly detected in Coptopsylla lamellifer ardua, Paradoxopsyllus repandus, and Nosopsyllus laeviceps laeviceps. Additionally, 3 Wolbachia endosymbionts were firstly found in X. gerbilli minax, X. conforms conforms, P. repandus, and N. laeviceps laeviceps. BLASTn analysis indicated 3 Bartonella species showed genotypic variation. Phylogenetic analysis revealed 3 Wolbachia endosymbionts were clustered into the non-Siphonaptera Wolbachia group. These findings extend our knowledge of the geographical distribution and carriers of B. rochalimae, B. grahamii, B. elizabethae, and Wolbachia spp. In the future, there is a need for China-Kazakhstan cooperation to strengthen the surveillance of flea-borne pathogens in wildlife.
Bartonella ; Ecosystem ; Lakes ; Rodentia ; Siphonaptera ; Wetlands ; Wolbachia ; Xenopsylla

Objective To understand the infected strains and prevalence of brucellosis in occupational population in Tacheng and Kashgar regions,Xinjiang Uygur Autonomous Region.Methods In September 2015,blood samples from occupational population (including herders,semi-agricultural and semi-pastoral,veterinarians) and non-occupational population (including students and cadres) were collected to detect Brucellaspecific antibody and bacterial nucleic acids by rose bengal plate test (RBPT),serological standard tube agglutination test (SAT) and PCR methods,respectively.The positive products of PCR were sent to Shanghai Sangon Biotechnology Co.,LTD.Then the sequence results were retrieved online using the basic alignment search tool (BLAST) in GenBank web page and uploaded to NCBI (National Center for Biotechnology Information).Results A total of 546 blood samples were tested,including 300 males,aged (55-± 15) years old,and 246 females,aged (54 ± 12) years old.The positive rates were 17.58％ (96/546) and 6.78％ (37/546) in 546 blood samples by serological method and genetic markers targeting omp22 and omp2,respectively.The positive rates were statistically significant (x2 =29.8,P ＜ 0.05).Additionally,based on BLAST analysis of outer membrane protein omp22 and omp2 genes,the positive products were identified as Brucella abortus,and the sequence similarity was 100.00％ (253/253,863/863 bp) to Brucella abortus strain Wisconsin genome assembly,chromosome (LT651712).Conclusions Brucellosis has a high infection rate in the occupational population of some animal husbandry-based groups in Xinjiang.The infection strain is abortive species Brucella,and health education for the occupational population and prevention of brucellosis should be strengthened to reduce the infection rate.

Objective: To observe the clinical efficacy and safety of transhepatic arterial chemoembolization (TACE) combined with radiofrequency ablation (RFA) in the treatment of colorectal cancer with liver metastasis. Methods: The data of 92 patients with colorectal cancer with liver metastasis admitted to Nanyang First People′s Hospital of Henan Province from January 2014 to January 2016 were retrospectively analyzed. A total of 46 patients treated with TACE were selected as the TACE group, and another 46 patients treated with TACE and RFA were selected as the combined group. The clinical efficacies of the two groups were compared, and the changes of Karnofsky functional status (KPS) scores before and after treatment in the two groups were analyzed. The incidences of complications in the two groups were calculated. Patients in the two groups were followed up, and the progress-free survival (PFS) and overall survival (OS) were calculated. Results: The disease control rate of the combined group was 82.61% (38/46), and that of the TACE group was 63.04% (29/46). The disease control rate of the combined group was higher than that of the TACE group (χ²=4.449, P=0.035). Before treatment, the KPS scores of the combined group and the TACE group were 71.84±4.37, 72.22±4.26, with no statistically significant difference (t=0.423, P=0.673). After treatment, the KPS scores of the two groups were higher than those before treatment, and the KPS score of the combined group was higher than that of the TACE group (79.81±6.15 vs. 75.86±6.02; t=3.108, P=0.003). The incidence of complications was 54.35% (25/46) in the combined group and 41.30% (19/46) in the TACE group. The difference between the two groups was not statistically significant (χ²=1.568, P=0.210). The median PFS and OS in the TACE group were 12.6 and 20.7 months, and those in the combined group were 18.9 and 28.2 months. The PFS and OS of the combined group were longer than those of the TACE group (χ²=72.025, P<0.001; χ²=26.580, P<0.001). Conclusion TACE combined with RFA is effective in the treatment of liver metastasis of colorectal cancer, which can effectively improve the KPS score of patients, prolong the PFS and OS, and do not increase the risk of complications.