OBJECTIVE: To investigate and analyze the storage temperatures of drugs for references of drug storage. METHODS: A classified statistical analysis was conducted on those drugs with temperature requirements in storage as instructed in drug package inserts. RESULTS: The stock drugs totaled 1 254 kinds,of which,483 (38.52%) had the item of temperature requirements in storage,158 with room temperature storage and 61 with cold storage had the item of specific temperature requirement. CONCLUSION: The storage temperature of drugs should be established based on the quality of different drugs so as to ensure the stability of drug quality. It is advisable to clarify the storage temperature in all the drug package inserts.

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.

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.

Background and purpose: Three-incision esophagectomy for patients with esophageal cancer has been increasingly used, but the incidence of either postoperative anastomotic leak or stricture is higher than that in intrathoracic gastroesophageal anastomosis. The purpose of this study was to investigate the clinical effects of the side-to-side stapled cervical gastroesophageal anastomosis in preventing anastomotic leak and stricture after three-incision esophagectomy. Methods: One hundred and twenty-seven patients undergoing three-incision esophagectomy for esophageal cancer with gastric replacement were reviewed. A side-to-side stapled cervical gastroesophageal anastomosis was performed in 71 cases and manually sewn anastomosis in 56 cases. The incidence of postoperative anastomotic leak and stricture was compared between these two groups and the results were statistically analyzed using SPSS11.0 soft.Results: Anastomotic leakage was noted in seven patients (9.9%) in the stapler group and fourteen patients (25.0%) in the manually sewn group (P=0.04). After the operation two patients (2.8%) in the stapler group and nine patients (16.1%) in the manually sewn group developed a benign esophageal stricture (P=0.02).The incidence of either postoperative anastomotic leak or stricture in cases of the stapler group was significantly lower than that in the manually sewn group (P<0.05). Conclusion: Construction of the cervical esophagogastric anastomosis with a side-to-side stapled technique provides a larger luminal diameter which significantly reduces the incidence of postoperative anastomotic strictures. The surgery also greatly reduces the incidence of anastomotic leaks and strictures, so it could be used as an alternative strategy for cervical esophagogastric anastomosis after esophagectomy for esophageal cancer

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 To summarize the experiences of Nuss procedure for complicated pectus excavatum.Methods From August 2006 to July 2011,443 patients with pectus excavatum received Nuss operation.Among them,95 patients with complicated pectus excavatum (CT Haller index ＞ 6) received modified technique of Nuss procedure,including multi-bar technique,oblique placed bar,double arc bar technique; fix with wire ; osteotomies ; auxiliary small incision; thoracoscopy placed by different lateral.Results All operations were successfully accomplished without severe complications.The mean operative time was (90.13 ± 39.12 ) minutes and the mean volume of blood loss was (45.41 ± 19.23 )ml.The mean hospital stay was (7.21 ± 2.87 ) days.All patients have been satisfied with their surgical correction.Therapeutic results evaluation was excellent in 75.8％ of patients,good in 16.8％.Conclusion Multi modified technique of Nuss procedure can lead to a satisfactory outcome for complicated pectus excavatum patients.

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.

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.

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.