Background/Aims: Four-week treatment of linvencorvir (RO7049389) was generally safe and well tolerated, and showed anti-viral activity in chronic hepatitis B (CHB) patients. This study evaluated the efficacy, safety, and pharmacokinetics of 48-week treatment with linvencorvir plus standard of care (SoC) in CHB patients. Methods: This was a multicentre, non-randomized, non-controlled, open-label phase 2 study enrolling three cohorts: nucleos(t)ide analogue (NUC)-suppressed patients received linvencorvir plus NUC (Cohort A, n=32); treatment-naïve patients received linvencorvir plus NUC without (Cohort B, n=10) or with (Cohort C, n=30) pegylated interferon-α (Peg-IFN-α). Treatment duration was 48 weeks, followed by NUC alone for 24 weeks. Results: 68 patients completed the study. No patient achieved functional cure (sustained HBsAg loss and unquantifiable HBV DNA). By Week 48, 89% of treatment-naïve patients (10/10 Cohort B; 24/28 Cohort C) reached unquantifiable HBV DNA. Unquantifiable HBV RNA was achieved in 92% of patients with quantifiable baseline HBV RNA (14/15 Cohort A, 8/8 Cohort B, 22/25 Cohort C) at Week 48 along with partially sustained HBV RNA responses in treatment-naïve patients during follow-up period. Pronounced reductions in HBeAg and HBcrAg were observed in treatment-naïve patients, while HBsAg decline was only observed in Cohort C. Most adverse events were grade 1–2, and no linvencorvir-related serious adverse events were reported. Conclusions 48-week linvencorvir plus SoC was generally safe and well tolerated, and resulted in potent HBV DNA and RNA suppression. However, 48-week linvencorvir plus NUC with or without Peg-IFN did not result in the achievement of functional cure in any patient.

Osteosarcoma (OS) stood as the most prevalent primary malignant bone tumor among children and adolescents, characterized by its aggressive nature and high propensity for metastasis, thus earning its classification as a highly malignant disease. The primary routes of metastasis in osteosarcoma encompassed hematogenous dissemination (the most common metastasis was lung metastasis) and lymph node involvement, with lymph node metastasis carrying a notably poorer prognosis when contrasted with lung metastasis. However, it was noteworthy that, at that time, clinical practice often overlooked the crucial aspect of conducting regional lymph node screening for newly diagnosed osteosarcoma patients, and there remained a dearth of standardized treatment protocols for osteosarcoma lymph node metastasis. The intricate mechanisms at the heart of osteosarcoma lymph node metastasis primarily revolved around the infiltration of lymphatic vessels by osteosarcoma cells possessing metastatic capabilities and the induction of lymphatic vessel formation by these cells. Nevertheless, it was imperative to underscore that our understanding of the comprehensive mechanisms underpinning the initiation and progression of these processes still remained incomplete. Moreover, following the adaptive proliferation of osteosarcoma cells within regional lymph nodes, there existed a complex biological process that involved a myriad of cytokines and signaling pathways. This process facilitated the metastasis of osteosarcoma cells to distant target organs, most notably the lungs, by virtue of the intimate anatomical relationships between the lymphatic and circulatory systems. Consequently, that comprehensive review sought to provide an all-encompassing exposition on various facets of osteosarcoma lymph node metastasis. These facets included the direct invasion of surrounding lymphatic vessels by osteosarcoma cells, the osteosarcoma cell-induced development of lymphatic vessels through the VEGFC/VEGFR-3 signaling axis, the remodeling of the lymph node microenvironment by osteosarcoma cells via Hsp5B and the Wnt/β-Catenin signaling pathway to ensure their adaptation and survival, the epithelial-mesenchymal transition of osteosarcoma cells promoting metastasis from lymph nodes to the lung, and the utilization of small-molecule compounds in the battle against osteosarcoma lymph node metastasis. The aim was to provide a comprehensive and systematic elucidation of the intricate mechanisms governing osteosarcoma lymph node metastasis and to furnish invaluable insights for the development of therapeutic strategies.

AIM: To observe the effect of PCSK9 inhibitors on blood lipid levels and common inflammatory factors in patients with coronary heart disease. METHODS: The clinical data of 201 patients with coronary heart disease who were admitted to the Department of Cardiology of Shanghai East Hospital from April 2020 to June 2021 were retrospectively analyzed. The patients were divided into PCSK9 inhibitor treatment group (101 cases: statin + PCSK9 inhibitor) and control group (100 cases: statin treatment only) according to their medication status. Blood lipids, blood routine, CRP and FIB were re-examined after 1 month of treatment. The changes of blood lipids and inflammatory factors before and after treatment were compared. RESULTS: Before treatment, there was no significant difference in blood lipids, blood routine, CRP and FIB between the two groups (P> 0.05). The levels of sdLDL and Lp(a) were significantly decreased (P< 0.05); the levels of WBC, CRP, N and FIB were significantly decreased (P< 0.05). Compared with the control group, the levels of TC, HDL, LDL-C, CRP and FIB in the PCSK9 inhibitor group were significantly changed (P< 0.05), and the results were statistically significant. CONCLUSION: PCSK9 inhibitors can not only reduce LDL-C levels, but also reduce Lp(a) levels. PCSK9 inhibitors can reduce CRP and FIB levels, suggesting that it can partially improve inflammation in peripheral blood in patients with coronary heart disease.

Objective To investigate the effect of Dexamethasone (Dex) on the proliferation and apoptosis of osteoblasts in vitro and to explore its underlying mechanism.Methods Osteoblasts were acquired by primary culturing from new born SD rats.The inverted microscope was used to observe the cellular appearance.The cells were identified by alkaline phosphatase staining and alizarin red staining.The third generation osteoblasts were divided into four groups.Cells were incubated with different concentrations (0,10-8 mol/L,10-7 mol/L,10-6 mol/L) of dexamethasone for 12 hours,24 and 48 hours.Cell Counting Kit-8 was performed to evaluate the inhibitory effect on cell proliferation.The apoptosis rate was analyzed by flow cytometry with Annexin Ⅴ-FITC/PI double staining.The fluorescence microscopy was used to test the nuclear alteration and the expression of caspase-3.Western blot assay was applied to detect the expression of Bcl-2,Bad,caspase-3 and phosphorylated Akt.One-Way analysis of variance was used to determine the difference between groups.LSD-t was used to compare the difference between any two groups.Results Com-pared with the control group,dexamethasone at dose of 10-8 mol/L,10-7 mol/L and 10-6 mol/L inhibited the proliferation of osteoblasts,most evidently in 48 hours (0.980±0.028 vs 1.143±0.017,t=5.454,P＜0.05;0.798±0.057 vs 1.143±0.017,t=1 1.555,P＜0.05;0.728±0.031 vs 1.143±0.017,t=13.908,P＜0.05).Dexamethasone at dose of 10-7 mol/L and 10-6 mol/L induced apoptosis of osteoblasts at 48 hours,showing significant difference compared with control group [(9.8± 2.6)％ vs (4.1±0.8)％,t=3.508,P＜0.05;(12.4±2.6)％ vs (4.1±0.8)％,t=5.140,P＜0.05].However,10-8 mol/L of dexamethasone had no apparent effect in inducing apoptosis of osteoblasts [(4.9±1.2)％ vs (4.1±0.8)％,t=0.470,P ＞0.05].The immunofluorescene staining result showed that the expression of caspase-3 protein was significantly increased in 10-7 mol/L and 10-6 mol/L dex group (t=4.320,8.475,P＜0.05).The Western blotting results showed that dexamethasoneat the concentration of 10-7 mol/L,10-6 mol/L could significantly increase the expression of Bad and caspase-3 and down-regulate the expression of Bcl-2 and p-Akt.The expression of Bcl-2 was markedly reduced by 53.8％,78.4％ (t=4.019,5.988;P＜0.05),The expression of p-Akt decreased by 37％,49.6％ (t=2.067,3.491;P＜0.05),the expression of Bad protein increased by 276.9％ and 334.8％ respectively (t=7.342,8.872;P＜0.05),the expression of caspase-3 protein were increased by 138.0％ and 193.9％ (t=5.510,7.750;P＜0.05).Conclusion Dexamethasone is capable of inhibiting the proliferation of osteoblast,as well as augmenting the apoptosis.The mechanism of this process is probably related to reduction of the level of Bcl-2 expressionand up-regulation the expression of Bad,caspase-3 with the effects of inhibiting the PI3K/Akt signaling pathway.

BACKGROUND:Osteoblasts with high purity and activity are essential for bone metabolism research. OBJECTIVE:To explore a simple and effective culturing method of primary osteoblasts. METHODS:Osteoblasts were isolated from the parietal and frontal bones of newborn Sprague-Dawley rats using trypsin and collagenase digestion and tissue explant method. The morphology of osteoblasts was observed by inverted phase contrast microscope and transmission electron microscope;the cells was counted to draw the growth curve;the osteoblasts were identified by alkaline phosphatase BCIP/NBT staining and alizarin red staining. RESULTS AND CONCLUSION:The cells showed spindle, triangle or polygon shapes, having two or three protrusions. There were abundant mitochondria and endoplasmic reticulum under electron microscope, which presented the typical characteristics of osteoblasts. The cell growth was slow intially, accelerating at the 3rd day, and peaking at the 7th day. The cells were highly positive for alkaline phosphatase staining and were stained orangered through the alizarin red staining. To conclude, the cells isolated using enzymatic digestion combined with tissue explant method exhibit the typical characteristics and functions of osteoblasts, and this method is an ideal way to culture primary osteoblasts.

Objective: To investigate the clinical effect and safety of long-acting pegylated interferon-α-2b (Peg-IFN-α-2b) (Y shape, 40 kD) injection (180 μg/week) in the treatment of HBeAg-positive chronic hepatitis B (CHB) patients, with standard-dose Peg-IFN-α-2a as positive control. Methods: This study was a multicenter, randomized, open-label, and positive-controlled phase III clinical trial. Eligible HBeAg-positive CHB patients were screened out and randomized to Peg-IFN-α-2b (Y shape, 40 kD) trial group and Peg-IFN-α-2a control group at a ratio of 2:1. The course of treatment was 48 weeks and the patients were followed up for 24 weeks after drug withdrawal. Plasma samples were collected at screening, baseline, and 12, 24, 36, 48, 60, and 72 weeks for centralized detection. COBAS® Ampliprep/COBAS® TaqMan® HBV Test was used to measure HBV DNA level by quantitative real-time PCR. Electrochemiluminescence immunoassay with Elecsys kit was used to measure HBV markers (HBsAg, anti-HBs, HBeAg, anti-HBe). Adverse events were recorded in detail. The primary outcome measure was HBeAg seroconversion rate after the 24-week follow-up, and non-inferiority was also tested. The difference in HBeAg seroconversion rate after treatment between the trial group and the control group and two-sided confidence interval (CI) were calculated, and non-inferiority was demonstrated if the lower limit of 95% CI was > -10%. The t-test, chi-square test, or rank sum test was used according to the types and features of data. Results: A total of 855 HBeAg-positive CHB patients were enrolled and 820 of them received treatment (538 in the trial group and 282 in the control group). The data of the full analysis set showed that HBeAg seroconversion rate at week 72 was 27.32% in the trial group and 22.70% in the control group with a rate difference of 4.63% (95% CI -1.54% to 10.80%, P = 0.1493). The data of the per-protocol set showed that HBeAg seroconversion rate at week 72 was 30.75% in the trial group and 27.14% in the control group with a rate difference of 3.61% (95% CI -3.87% to 11.09%, P = 0.3436). 95% CI met the non-inferiority criteria, and the trial group was non-inferior to the control group. The two groups had similar incidence rates of adverse events, serious adverse events, and common adverse events. Conclusion In Peg-IFN-α regimen for HBeAg-positive CHB patients, the new drug Peg-IFN-α-2b (Y shape, 40 kD) has comparable effect and safety to the control drug Peg-IFN-α-2a.

Objective To study the influence of the minimal invasive Nuss procedure on the pulmonary function of post-operative pectus excavatum patients. Methods Conduct retrospective analysis on the data from 676 pectus excavatum patients who were treated by the minimal invasive Nuss procedure from August 2006 to November 2014. Wherein 182 cases have com-plete preoperative and postoperative pulmonary-function data of one year, three years. These cases were divided into 3 groups according to the age, namely, children group( from 6 to 12 years old, 34 cases) , adolescents group( from 13 to 18 years old, 80 cases) , adults group( above 18 years old, 68 cases) , among which there were 71 cases with pulmonary function data of 1 year after removal of steel plate, they were divided into 3 groups in the same way,namely, children group(20 cases), adoles-cents group(22 cases), adults group(29 cases) . To compare and analyze the pulmonary function indicatrix of patients with dif-ferent ages in preoperative stage, 1 year, 3 years postoperative stages and 1 year after dismantling the steel plate stage, and to investigate the influence of the minimal invasive Nuss procedure on the postoperative lung function. Results The pulmonary function indicatrix in preoperative stage, 1 year, 3 years postoperative stages of the children group did not have significant difference(P>0. 05); the FVC, FEV1 indicatrix of adolescents and adults groups declined after operation in 1 year and 3 years compared with the preoperative stage(P<0. 05), FEF 25％ -75％, FEF 50％, FEF75％ were improved after operation in 1 year and 3 years compared with the preoperative stage(P<0. 05);the pulmonary function indicatrix of three age groups in the 1 year after dismantling the steel plate stage had all improved, in which the pulmonary function indicatrix of the children group improves most significantly(P<0. 05). Conclusion After the minimal invasive Nuss procedure before the plate dis-mantling process, the pulmonary function of children patients remains to be similar. Partial ventilatory function was damaged in the adolescents and adults patients. After the plate dismantling process, the pulmonary function indicatrix of each age group hasimproved in different degrees. Improvement effect is the most significant in patients below the age of 12.

The author carried on bibliometrics statistical analysis of published papers during 2003-2012 by Disease control agencies of Chongqing and put forward some suggestions based on the results of the analysis.The results showed that in 10 years,the number of published papers for a total of 1154 with an average annual growth rate of 10.41％,of which 467 core journals,journal article 687.the difference between the municipal,district and county larger.The quantity and quality of published papers showed an upward trend,but there is still development space and the quality of the paper should be improved.

Objective To establish a novel and convenient method to study the phenotype of drug resistant hepatitis B virus (HBV) isolates,and to analyze the drug susceptibility by replacing the reverse transcriptase (RT) domain of wild-type HBV with that of the drug resistant HBV isolates.Methods Full length of HBV isolates was amplified and cloned from the sera of patients prior to nucleoside/nucleotide analogues (NA) treatment.Wild-type full-length HBV genomes was used to construct the recombinant expression plasmids PHY536207 (genotype B) and PHY97 (genotype C).The restriction enzyme sites were introduced in the upstream and downstream region of reverse transeription (RT) domain to construct plasmid,which were named as mPHY536207 and mPHY97,respectively.Lamivudine (LAM) resistant mutant and adefovir (ADV) resistant mutant were isolated and cloned to construct recombinant expression plasmids PHY634 and PHY6923,respectively.Subsequently,the RT domain of mPHY536207 was replaced by that of drug resistant mutant to construct the plasmids RT634 (LAM-resistant) and RT6923 (ADVresistant).The HBV constructs were transfected into Huh7 cells.The HBsAg levels in supernatant were determined by enzyme-linked immunosobent assay (ELISA),and the amount of intracellular HBV DNA was assayed by real-time polymerase chain reaction and Southern blot analysis.Results The plasmids PHY536207 and PHY97 containing genotype B and genotype C wild-type fulllength HBV genomes were constructed successfully,both of which could replicate in Huh7 cells.Intracellular HBV DNA extracted from cells in each of six-well culture plates was more than 1 × 107 copy/ mL,and the introduction of Pst Ⅰ restriction enzyme site did not affect the viral replication and HBsAg secretion.PHY634 and RT634,in which mutant RT domain was replaced into a wild type HBV expressing vector,exhibited the same HBV DNA replication under the treatment with different doses of LAM,the value of 50％ inhibitory concentration (IC50) was ＞100 μmol/L,while the IC50 of mPHY536207 was 0.18μmol/L.Moreover,wild-type isolate was sensitive to ADV (IC50 =1.2 μmol/L),while PHY6923 and RT6923 were resistant to ADV treatment (IC50 ＞100 μmol/L).Conclusion The phenotypic assay is successfully developed in this study based on replacing RT domain of wild-type HBV strains with that of clinical isolated drug resistant strain.

Objective The Nuss procedure is a minimally invasive pectus repair.The aim of this study was to explore the changes in quality of life in patients who underwent a Nuss procedure.Methods 96 patients,who underwent a Nuss procedure in our institution,were interviewed at preoperation,1 year after operation with the bar in place,and before the planned bar removal.The Nuss Questionnaire modified by George Krasopoulos et al,which evaluates psychosocial and physical well-being,was independently used by patients.Results Most of the scoring of the individual questions and the total score of individual patients revealed a significant improvement,both in preoperation vs.1 year after operation and 1 year after operation vs.before the planned bar removal.Conclusion The Nuss procedure has been shown to improve the quality of life in patients with pectus excavatum deformity in the short term.