Etiological analysis of acute respiratory tract infection in Taizhou of Zhejiang Province from 2020 to 2021 / 上海预防医学

ObjectiveWe examined the principal respiratory pathogens in patients with acute respiratory tract infection in Taizhou， Zhejiang Province during 2020‒2021 to provide evidence for prevention， diagnosis and treatment of acute respiratory tract infection. MethodsFrom September 2020 to August 2021， a total of 2 831 cases with acute respiratory tract infection were collected from two influenza sentinel surveillance hospitals in Taizhou， which had then received the examination of 22 respiratory pathogens by multiple fluorescence quantitative PCR. ResultsThe total positive rate of respiratory pathogens in 2 831 samples was 14.13%， among which enterovirus （7.77%） and respiratory syncytial virus （1.59%） were the principal pathogens. Except enterovirus， there was no significant difference in the positive rate of pathogens detected by gender（P>0.05）. Moreover， there was significant difference in pathogens by age （P<0.05）， with the highest positive rate in 0‒4 years（35.21%）. There was also significant difference in pathogens by seasons （P<0.05）， with the highest positive rate in summer（20.54%）. ConclusionThe positive rate of acute respiratory tract infection decreases significantly， compared with that before the COVID-19 epidemic. The differences in the positive rate differ significantly by age and seasons. Comprehensive consideration of diverse factors before diagnosis and the utilization of multiple fluorescent quantitative PCR can quickly and effectively determine the pathogens in the early stage of infection. Our findings may provide certain support for the diagnosis and treatment of acute respiratory infections in the context of COVID-19 in Taizhou.

Genotyping and molecular tracing of the first local infection of dengue virus in Taizhou / 中华实验和临床病毒学杂志

Objective@#To determine the first dengue fever case in Taizhou and trace probable transmission sources.@*Methods@#Collected serum of three patients for antigen, antibody and nucleic acid detection. Dengue viruses were isolated and cultured in C6/36 cell. The whole length of E gene was amplified by reverse transcriptase-polymerase chain reaction(RT-PCR) and then sequenced. The phylogenetic tree was drawn.@*Results@#Three cases were positive in nucleic acid detection. Two cases were IgM positive. One case was NSI antigen postive. Three strains of type I dengue virus were isolated from samples. The phylogenetic trees shown that the three strains were on the same branch. The identities of nucleotide were 99.87%. The identities of amino acid were 99.6%-99.8%.@*Conclusions@#The dengue virus strains isolated in Taizhou was imported from Guangdong or Southeast Asia and caused location infection.

Expression of sterol regulatory element-binding protein-2 in the process of chondrocyte degeneration / 中国组织工程研究

BACKGROUND: Recent studies have demonstrated that sterol regulatory element binding protein-2 (SREBP-2) plays a key role in osteoarthritis, but its exact pathogenesis remains incompletely understood yet. OBJECTIVE:To investigate the expression of SREBP-2 in the process of interleukin-1β-induced articular chondrocyte degenerationin vitro. METHODS: Articular chondrocytes obtained from C57BL/6J mice were culturedin vitro. After the second passage, cels were randomly divided into four groups: control group, and three experimental groups treated with 10 μg/L interleukin-1β for 24, 48 and 72 hours, respectively. RESULTS AND CONCLUSION:The cels became hypertrophic after being stimulated by interleukin-1β, and the staining of colagen X was positive at 72 hours. MTT assay demonstrated that the cel activity after stimulation with interleukin-1β decreased with time. Results of RT-PCR showed that the expression of SREBP-2 and SREBP cleavage activating protein mRNA was significantly increased after stimulation with interleukin-1βas compared with the control group and increased with time. On the contrary, the expression of aggrecan and colagen II mRNA was decreased with time. It is revealed that interleukin-1β could inhibit the proliferation of regular chondrocytes and the expression of its extracelular matrix, and furthermore, induce chondrocyte hypertrophy. The expression of SREBP-2 showed a negative relationship with key cartilage genes during this interleukin-1β-induced degeneration.

Influences of IL-6R antibody on PMMA bone cement-mediated expression of OPG and RANKL in synovial fibroblasts / 华中科技大学学报（医学）（英德文版）

Effect of interleukin-6 receptor (IL-6R) antibody on polymethyl methacrylate (PMMA) bone cement-mediated expression of osteoprotegerin (OPG) and receptor activator of nuclear factor-kappaB ligand (RANKL) in synovial fibroblasts was investigated. Synovial tissue obtained from total knee arthroplasty was digested and cultured. Inverted microscope was employed to observe the synovial cells and immunocytochemistry (SABC method) staining was used to identify synovial fibroblasts. This experiment was divided into three groups according to different culture media: PMMA group (75 μg/mL PMMA bone cement particles), IL-6R antibody group (10 ng/mL IL-6R antibody+75 μg/mL PMMA bone cement particles), and control group (no IL-6R antibody or PMMA bone cement particles). Influence of IL-6R antibody and PMMA on proliferation of synovial fibroblasts was measured by cell counting kit-8 (CCK-8). ELISA method was used to measure OPG and RANKL levels in culture solution. Fluorescence quantitative real-time PCR (FQ-PCR) was used to detect the expression of OPG and RANKL mRNA. After three consecutive passages, more than 95% of the primary synovial cells became long spindle fibroblast-like cells. SABC staining results showed that the fibroblast-like cells were negative for anti-CD68 antibody and positive for anti-vimentin antibody, with brown madder stained. CCK-8 test demonstrated that the absorbance (A) value at 450 nm was significantly lower in IL-6R antibody group than in PMMA group and control group (P0.05). Results of ELISA indicated that the expression of OPG was significantly higher in IL-6R antibody group than in PMMA group and control group (P0.05), but the expression of RANKL was higher in PMMA group than in control group (P0.05). IL-6R antibody could significantly increase the expression of OPG, but inhibit the expression of RANKL, which might provide a theoretical basis of molecular biology for the prevention and treatment of aseptic loosening of prosthesis.

Influences of IL-6R antibody on PMMA bone cement-mediated expression of OPG and RANKL in synovial fibroblasts / 华中科技大学学报（医学）（英德文版）

Effect of interleukin-6 receptor (IL-6R) antibody on polymethyl methacrylate (PMMA) bone cement-mediated expression of osteoprotegerin (OPG) and receptor activator of nuclear factor-kappaB ligand (RANKL) in synovial fibroblasts was investigated. Synovial tissue obtained from total knee arthroplasty was digested and cultured. Inverted microscope was employed to observe the synovial cells and immunocytochemistry (SABC method) staining was used to identify synovial fibroblasts. This experiment was divided into three groups according to different culture media: PMMA group (75 μg/mL PMMA bone cement particles), IL-6R antibody group (10 ng/mL IL-6R antibody+75 μg/mL PMMA bone cement particles), and control group (no IL-6R antibody or PMMA bone cement particles). Influence of IL-6R antibody and PMMA on proliferation of synovial fibroblasts was measured by cell counting kit-8 (CCK-8). ELISA method was used to measure OPG and RANKL levels in culture solution. Fluorescence quantitative real-time PCR (FQ-PCR) was used to detect the expression of OPG and RANKL mRNA. After three consecutive passages, more than 95% of the primary synovial cells became long spindle fibroblast-like cells. SABC staining results showed that the fibroblast-like cells were negative for anti-CD68 antibody and positive for anti-vimentin antibody, with brown madder stained. CCK-8 test demonstrated that the absorbance (A) value at 450 nm was significantly lower in IL-6R antibody group than in PMMA group and control group (P<0.01), but there was no statistically significant difference in A value at 450 nm between the control group and PMMA group (P>0.05). Results of ELISA indicated that the expression of OPG was significantly higher in IL-6R antibody group than in PMMA group and control group (P<0.01). The expression of RANKL was inhibited (P<0.05), and the ratio of OPG/RANKL was significantly increased in IL-6R antibody group as compared with PMMA group and control group. There was no significant difference in the expression of OPG between control group and PMMA group (P>0.05), but the expression of RANKL was higher in PMMA group than in control group (P<0.05), and there was a significant difference in the ratio of OPG/RANKL between them (P<0.05). Results of FQ-PCR revealed the expression of RANKL mRNA was significantly inhibited (P<0.01) and the expression of OPG mRNA was significantly increased (P<0.01) in IL-6R antibody group as compared with PMMA group and control group. The expression of RANKL mRNA was higher in PMMA group than in control group (P<0.05), but the expression of OPG mRNA had no significant difference between them (P>0.05). IL-6R antibody could significantly increase the expression of OPG, but inhibit the expression of RANKL, which might provide a theoretical basis of molecular biology for the prevention and treatment of aseptic loosening of prosthesis.

Liver histopathological features influencing HBeAg seroconversion in patients with HBeAg-positive chronic hepatitis B responding to Peg-IFN treatment / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the therapeutic efficiency of antiviral treatment with pegylated-interferon (Peg-IFN) for hepatitis B e antigen (HBeAg)-positive chronic hepatitis B (CHB) and to explore whether liver histopathological features or other factors influence the HBeAg seroconversion treatment response.</p><p><b>METHODS</b>Eighty HBeAg-positive CHB patients with diagnosis confirmed by liver puncture were treated with Peg-IFN(2a or 2b)body weight dose, once weekly). At treatment week 48, the rate of HBeAg seroconversion was determined and used to analyze the influence of liver histopathological features (liver biopsy assessment of: inflammation, graded G0 to G4; fibrosis stage, graded S0 to S4), sex, age, differential levels (pre-treatment baseline vs. week 48 post-treatment) of serum alanine transferase (ALT), and HBV DNA, by binary logistic analysis.</p><p><b>RESULTS</b>At week 48, the overall rate of HBeAg seroconversion was 30.0%. The rate of HBeAg seroconversion gradually advanced with increased liver inflammation (X2 = 8.435, P = 0.015): 9.09% of the 22 patients with G1; 31.58% of the 38 patients with G2; 47.30% of the 19 patients with G3; the one patient with G4. In contrast, the rate of HBeAg seroconversion showed a much weaker association with liver fibrosis (X2 = 5.917, P = 0.116). Only baseline HBeAg level, and no other baseline index, was significantly different between the patients who achieved HBeAg seroconversion and those who did not. Liver inflammation and baseline HBeAg level were identified as influencing factors of HbeAg seroconversion in response to Peg-IFN treatment.</p><p><b>CONCLUSION</b>Peg-IFN therapy induces a higher rate of HBeAg seroconversion in HBeAg-positive CHB patients with severe liver inflammation; histological analysis of pre-treatment liver biopsies may help to identify patients most likely to benefit from the antiviral regimen.</p>

Study of the effect and mechanism of spastic paraplegia 21 protein on the replication of hepatitis B virus / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the effect of human spastic paraplegia 21 protein (SPG21) on the replication of hepatitis B virus(HBV) and its regulatory mechanism.</p><p><b>METHODS</b>HBV infectious clone pHBV1.3 and its promoter pHBV-Luc were transfected respectively into HepG2 cells with SPG21 of different concentrations, HBsAg and HBeAg in the supernatants were measured by enzyme linked immunosorbent assay (ELISA), expression of HBV core mRNA and protein were detected by RT-PCR and western blot, covalently closed circular DNA(ccc DNA) levels were measured by real-time PCR, and HBV promoter activity was measured by luminometer fluorescence detector.</p><p><b>RESULTS</b>Expression of HBsAg, HBeAg, HBV core protein and cccDNA were upregulated by SPG21 as well as HBV promoter activity in a dose-dependent approach. The activity of HBV promoter increased to 1.63, 3.09 and 4.66 times in HepG2 cells treated with 50mug/ml, 100mug/ml and 200mug/ml SPG21 respectively during 48 hour-treated ( P less than 0.05), as compared to the control group.</p><p><b>CONCLUSIONS</b>SPG21 can enhance the replication of HBV in HepG2 cells.</p>

Effect of hepatitis B virus X gene on the expression of spastic paraplegia 21 / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effect of hepatitis B virus(HBV) X gene on the expression of SPG21.</p><p><b>METHODS</b>The expressions of SPG21 mRNA and protein in HepG2 and HepG2.2.15 cells were tested by RT-PCR and western blot. HepG2 cells were co-transfected with reporter plasmid pGL3-SPG21 and plasmids carrying individual genes of HBV, the luciferase activity was measured and the expressions of SPG21 were detected by RT-PCR and western blot.</p><p><b>RESULTS</b>The expressions of SPG21 mRNA and protein were higher in HepG2.2.15 cells than in HepG2 cells (0.36+/-0.06 vs 0.21+/-0.05, P value is less than 0.05). The activity of SPG21 in HepG2 cells transfected with pCMV-X was higher (875+/-27 vs 67+/-12, P value is less than 0.01) as compared to blank control group (transfected with pCMV-tag2B). HBV X gene enhanced SPG21 gene promoter activity, SPG21 mRNA expression and SPG21 protein production in HepG2 cells in a dose-dependent manner.</p><p><b>CONCLUSION</b>HBV X gene can specially activate SPG21 expression.</p>

Serum levels of leptin and several growth-related hormones in premature infants / 中华儿科杂志

<p><b>OBJECTIVE</b>Leptin (LEP) is mainly produced by white adipose tissue and participates in the energy metabolism and regulation of growth. Cooperating with the other metabolic hormones, it plays an important role in the developments of fetus and neonates. This study was designed to test the serum levels of LEP, neuropeptide Y (NPY), insulin (INS) and insulin-like growth factor-1 (IGF-1) and measure the body mass index (BMI) and head circumference (HC) at different days of life of premature infants with or without serious diseases and to find the changes of serum levels of LEP as well as NPY, INS and IGF-1, the relationship between those hormones and the changes of body weight and the influences of diseases on the levels of those hormones in premature infants.</p><p><b>METHOD</b>The clinical data as well as weights, lengths, HC of 40 sick premature infants (sick group) and 30 premature infants without any diseases (control group) were collected and the serum levels of LEP, NPY, INS and IGF-1 were determined by using radioimmunoassay (RIA) at d 1, d 7 and d 12 of life. BMI was calculated by weight (kg)/length (m)(2). SPSS13.0 was used to analyze the data</p><p><b>RESULT</b>(1) In sick group the serum LEP levels were 0.74 +/- 0.21, 0.60 +/- 0.18, 0.82 +/- 0.12 (mg/L) (P < 0.01), the BMI were 9.81 +/- 1.24, 8.36 +/- 0.87, 9.08 +/- 1.12 (kg/m(2)) (P < 0.01) on d 1, d 7 and d 12, respectively. In control group serum LEP levels were 0.78 +/- 0.17, 0.71 +/- 0.17, 0.88 +/- 0.58 (mg/L) (P < 0.01), the BMI were 10.03 +/- 1.04, 9.35 +/- 0.80, 11.06 +/- 0.82 (kg/m(2)), on d 1, d 7 and d 12, respectively (P < 0.01). In both groups, serum LEP levels as well as the BMI decreased on d 7 and reincreased on d 12. The differences of serum LEP levels and BMI between sick group and control group at d1 were not significant (P > 0.05); compared with control group, the serum LEP levels and BMI on d 7 and d 12 in sick group were lower and the differences were significant. (2) There were positive correlations between serum LEP levels and BMI in sick group as well as in control group. (3) In sick group, the serum NPY levels at d 1, d 7, d 12 were 55.33 +/- 9.38, 46.64 +/- 6.17, 75.13 +/- 9.12 (ng/L) (P < 0.01), INS were 10.07 +/- 2.63, 7.71 +/- 2.77, 10.37 +/- 2.29 (mU/L) (P < 0.01), IGF-1 were 38.66 +/- 11.42, 31.98 +/- 7.34, 41.84 +/- 8.05 (mg/L) (P < 0.01), respectively. In control group, the serum NPY levels at d1, d 7 and d 12 were 57.77 +/- 7.15, 48.49 +/- 8.81, 81.36 +/- 8.51 (ng/L) (P < 0.01), INS were 11.55 +/- 1.99, 8.28 +/- 2.87, 15.42 +/- 3.80 (mU/L) (P < 0.01), IGF-1 were 37.76 +/- 7.07, 34.33 +/- 8.97, 50.19 +/- 8.38 (mg/L) (P < 0.01), respectively. In both groups, serum levels of NPY, INS and IGF-1 had positive correlations with serum LEP levels as well as BMI on the corresponding days and decreased on d 7 and reincreased on d 12.</p><p><b>CONCLUSION</b>(1) The serum LEP levels decreased on 7 d of life and reincreased on 12 d of life, which corresponded to the changes of the physical development of premature infants. (2) The serum LEP levels in sick premature infants decreased definitely as compared with control group, which suggested that diseases had negative influences on the LEP levels and the physical developments were slowed down in sick premature infants. (3) The serum levels of NPY, INS and IGF-1 had positive correlations with LEP levels as well as BMI at the early period of life, which suggested that NPY, INS and IGF-1, cooperating with LEP, might take part in the regulation of development of premature infants.</p>

Relationships among magnetic resonance imaging, histological findings, and IGF-I in steroid-induced osteonecrosis of the femoral head in rabbits / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

<p><b>OBJECTIVE</b>To study the relationships among magnetic resonance imaging (MRI), histological findings, and insulin-like growth factor-I (IGF-I) in steroid-induced osteonecrosis of the femoral head in rabbits.</p><p><b>METHODS</b>Thirty rabbits were randomly divided into experimental Group A (n=15) and control Group B (n=15). The 7.5 mg/kg (2 ml) of dexamethasone (DEX) and physiological saline (2 ml) were injected into the right gluteus medius muscle twice at one-week intervals in animals of Groups A and B, respectively. At 4, 8 and 16 weeks after obtaining an MRI, the rabbits were sacrificed and the femoral head from one side was removed for histological study of lacunae empty of osteocytes, subchondral vessels, and size of fat cells under microscopy, and the femoral head from the other side was removed for enzyme-linked immunoadsorbent assay (ELISA) for IGF-I.</p><p><b>RESULTS</b>At 4, 8 and 16 weeks after treatment, no necrotic lesions were detected in Group B, while they were detected in Group A. Light microscopy revealed that the fat cells of the marrow cavity were enlarged, subchondral vessels were evidently decreased, and empty bone lacunae were clearly increased. The IGF-I levels in Group A were significantly higher than those in Group B. At 8 weeks after the DEX injection, the MRI of all 20 femora showed an inhomogeneous, low signal intensity area in the femoral head, and at 16 weeks, the findings of all 10 femora showed a specific "line-like sign". The MRI findings of all femora in Group B were normal.</p><p><b>CONCLUSION</b>MRI is a highly sensitive means of diagnosing early experimental osteonecrosis of the femoral head. However, the abnormal marrow tissues appeared later than 4 weeks when the expression of IGF-I increased. This reparative factor has an early and important role in response to steroid-induced osteonecrosis of the femoral head, and provides a theoretical foundation for understanding the pathology and designing new therapies.</p>

An analysis of CD3+CD56+ lymphocytes and their subsets in the peripheral blood of patients with chronic hepatitis B / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To investigate CD3+CD56+ lymphocytes and their subsets in the peripheral blood of chronic hepatitis B patients and to explore the relationship between these cells and the pathogenesis of their diseases.</p><p><b>METHODS</b>Blood samples from 53 chronic hepatitis B patients, 17 from HBV asymptomatic carriers (ASC) and 19 from healthy controls (HC) were collected. CD3+CD56+ lymphocytes were detected by flow cytometry (FCM), then the CD3+CD56+ lymphocytes were gathered to analyze their expressions of CD4, CD8, TCR Valpha24, TCRalpha/beta and TCRgamma/delta.</p><p><b>RESULTS</b>The number of CD3+CD56+ lymphocytes of chronic hepatitis B patients (7.4+/-4.6%) was more than those of ASC (4.5%+/-3.5%) and healthy controls (4.4%+/-3.7%). The expressions of TCR Valpha24 on CD3+CD56+ lymphocytes showed no significant differences among the three groups, but the expression of TCR Valpha24 on CD3-CD56+ lymphocytes of ASC ( 2.8%+/-1.4% ) was much more than that of the HC (1.7%+/-1.0%). For the subsets analysis, the CD8 and TCRalpha/beta subsets of CD3+CD56+ lymphocytes of chronic hepatitis B (61.9%+/-16.8% and 68.1%+/-16.9%) were significantly higher than those of the HC (49.2%+/-15.6% and 56.4%+/-17.9%), while the TCRgamma/delta subsets of chronic hepatitis B and ASC (29.6%+/-15.4% and 30.5%+/-14.8%) were decreased significantly than those of the HC (41.4%+/-19.4%). On the other hand, the CD8 and TCRalpha/beta subsets of CD3+CD56+ lymphocytes of severe chronic hepatitis B (69.0%+/-14.0% and 76.1%+/-12.9%) and CD8 subsets of moderate chronic hepatitis B patients (66.4%+/-14.9%) were significantly higher than those of the mild chronic hepatitis B patients (51.4%+/-16.2% and 62.1%+/-14.6%).</p><p><b>CONCLUSION</b>The pathogenesis of chronic hepatitis B may positively relate to the high expression of CD8 on the CD3+CD56+ lymphocytes.</p>

Treatment of hepatocellular carcinoma in mice with locally administered epirubicin-loaded poly(D, L)-lactic acid microspheres / 中国医学科学院学报

<p><b>OBJECTIVE</b>To study the effectiveness of treating hepatocellular carcinoma (HCC) in mice with locally administered epirubicin-loaded poly( D, L) - lactic acid microspheres (EPI-PLA-MS ).</p><p><b>METHODS</b>EPI-PLA-MS was prepared with double emulsion solvent evaporation technique. Five groups of mice (n = 8 in each group) were intraperitoneally injected with five different doses of free epirubicin (FEPI), and the maximum tolerated dose (MTD) was calculated. Then 15 mice with transplanted subcutaneous H22 HCC were divided into three groups (n = 5), which were respectively intratumorally injected with normal saline (NS), blank microspheres, and EPI-PLA-MS (with 9 mg/kg of EPI). After two weeks the tumors were excised and weighed. Another 15 mice with transplanted H22 ascites HCC were divided into three groups (n = 5), which were intraperitonealy injected with the same drugs, and the increased life span were registered exactly.</p><p><b>RESULTS</b>The MTD of intraperitoneally injected FEPI was 9 mg/kg. The tumour-inhibiting rates was 40.35% and 36.09% when EPI-PLA-MS were administered by intratumoral injection to the mice with subcutaneous H22 HCC. It significantly prolonged the survival time of mice with H22 ascites HCC and the increased life span by 153.49% and 142.22% when EPI-PLA-MS were intraperitoneally administered.</p><p><b>CONCLUSION</b>EPI-PLA-MS is a new sustained-release preparation with high-efficacy and low-toxicity in treating HCC and has shown promising prospects when administered locally.</p>