Advances in laboratory testing methods and strategies for hepatitis C virus infection / 临床肝胆病杂志

Laboratory testing for hepatitis C virus （HCV） infection provides an important basis for the identification and diagnosis of patients with HCV infection. With the continuous development of HCV testing in recent years， the performance of reagents has been significantly improved， and new testing service strategies have emerged and gradually been applied in clinical practice. This article summarizes the laboratory testing methods and strategies for HCV infection in China and globally， as well as the testing methods for HCV infection， and analyzes the influence of new methods and strategies on the prevention and control of HCV infection in China. Timely and accurate laboratory testing methods and effective and feasible testing strategies may help to realize the early identification， early diagnosis， and early treatment of HCV infection and ultimately achieve the strategic goal of eliminating viral hepatitis as a major public health threat by 2030.

Recombinant Schistosoma japonicum egg ribonuclease SjCP1412 inhibits the activation of LX-2 hepatic stellate cells in vitro / 中国血吸虫病防治杂志

Objective To investigate the effect of recombinant Schistosoma japonicum egg ribonuclease SjCP1412 (rSjCP1412) on proliferation, cell cycle, apoptosis and activation of human hepatic stellate cells LX-2 in vitro, and explore the underlying mechanisms. Methods The rSjCP1412 protein was expressed in Escherichia coli BL21 by prokaryotic expression, and the highly purified soluble rSjCP1412 protein was prepared by Ni NTA affinity chromatography and urea gradient refolding dialysis. Yeast RNA was digested using 12.5, 25.0, 50.0 µg rSjCP1412 proteins at 37 °C for 2, 3, 4 h, and the enzymatic products were electrophoresed on 1.5% agarose gel to observe the RNAase activity of rSjCP1412 protein. The proliferation of LX-2 cells stimulated by different doses of rSjCP1412 protein for 48 hours was measured using CCK-8 assay, and the apoptosis of LX-2 cells stimulated by different doses of rSjCP1412 protein for 48 hours was detected using the Annexin V-FITC/PI double staining, while the percentage of LX-2 cells at G0/G1, S and G2/M phases of cell cycle following stimulation with different doses of rSjCP1412 protein for 48 h was detected by DAPI staining. The type I collagen, type III collagen and α-smooth muscle actin (α-SMA) mRNA expression was quantified using quantitative florescent real-time PCR (qPCR) assay and Western blotting at transcriptional and translational levels in LX-2 cells following stimulation with different doses of rSjCP1412 protein for 48 h, while soluble egg antigen (SEA) served a positive control and PBS without rSjCP1412 protein as a normal control in the above experiments. The expression of collagen I, α-SMA and Smad4 protein was determined using Western blotting in LX-2 cells following stimulation with rSjCP1412 protein, transforming growth factor-β1 (TGF-β1) alone or in combination, to examine the signaling for the effect of rSjCP1412 protein on LX-2 cells. Results The rSjCP1412 protein was successfully expressed and the highly purified soluble rSjCP1412 protein was prepared, which had a RNase activity. Compared with the normal group, the survival rates of LX-2 cells significantly decreased post-treatment with 12.5, 25.0, 50.0 µg/mL rSjCP1412 protein and SEA for 48 h (F = 22.417 and 20.448, both P values < 0.05). The apoptotic rates of LX-2 cells significantly increased post-treatment with 12.5, 25.0, 50.0 µg/mL rSjCP1412 protein for 48 h (F = 11.350, P < 0.05), and treatment with 12.5, 25.0, 50.0 µg/mL rSjCP1412 protein for 48 h resulted in arrest of LX-2 cells in G0/G1 phase (F = 20.710, P < 0.05). Treatment with 12.5, 25.0, 50.0 µg/mL rSjCP1412 protein for 48 h caused a significant reduction in relative expression levels of collagen I (F = 11.340, P < 0.05), collagen III (F = 456.600, P < 0.05) and α-SMA mRNA (F = 23.100, P < 0.05) in LX-2 cells, and both rSjCP1412 protein and SEA treatment caused a significant reduction in collagen I (F = 1 302.000, P < 0.05), α-SMA (F = 49.750, P < 0.05) and Smad4 protein expression (F = 52.420, P < 0.05) in LX-2 cells. In addition, rSjCP1412 protein treatment inhibited collagen I (F = 66.290, P < 0.05), α-SMA (F = 31.300, P < 0.05) and Smad4 protein expression (F = 27.010, P < 0.05) in LX-2 cells activated by TGF-β1. Conclusion rSjCP1412 protein may induce apoptosis of LX-2 cells and inhibit proliferation, cell cycle and activation of LX-2 cells through down-regulating Smad4 signaling molecules.

Durable natural killer cell response after three doses of SARS-CoV-2 inactivated vaccine in HIV-infected individuals / 中华医学杂志(英文版)

BACKGROUND@#Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccine can induce a potent cellular and humoral immune response to protect against SARS-CoV-2 infection. However, it was unknown whether SARS-CoV-2 vaccination can induce effective natural killer (NK) cell response in people living with human immunodeficiency virus (PLWH) and healthy individuals.@*METHODS@#Forty-seven PLWH and thirty healthy controls (HCs) inoculated with SARS-CoV-2 inactivated vaccine were enrolled from Beijing Youan Hospital in this study. The effect of SARS-CoV-2 vaccine on NK cell frequency, phenotype, and function in PLWH and HCs was evaluated by flow cytometry, and the response of NK cells to SARS-CoV-2 Omicron Spike (SARS-2-OS) protein stimulation was also evaluated.@*RESULTS@#SARS-CoV-2 vaccine inoculation elicited activation and degranulation of NK cells in PLWH, which peaked at 2 weeks and then decreased to a minimum at 12 weeks after the third dose of vaccine. However, in vitro stimulation of the corresponding peripheral blood monocular cells from PLWH with SARS-2-OS protein did not upregulate the expression of the aforementioned markers. Additionally, the frequencies of NK cells expressing the activation markers CD25 and CD69 in PLWH were significantly lower than those in HCs at 0, 4 and 12 weeks, but the percentage of CD16 + NK cells in PLWH was significantly higher than that in HCs at 2, 4 and 12 weeks after the third dose of vaccine. Interestingly, the frequency of CD16 + NK cells was significantly negatively correlated with the proportion of CD107a + NK cells in PLWH at each time point after the third dose. Similarly, this phenomenon was also observed in HCs at 0, 2, and 4 weeks after the third dose. Finally, regardless of whether NK cells were stimulated with SARS-2-OS or not, we did not observe any differences in the expression of NK cell degranulation markers between PLWH and HCs.@*CONCLUSION@#s:SARS-CoV-2 vaccine elicited activation and degranulation of NK cells, indicating that the inoculation of SARS-CoV-2 vaccine enhances NK cell immune response.

Distributive characteristics of HBV DNA CpG islands in HBsAg positive mothers and its relationship with intrauterine transmission / 中华流行病学杂志

Objective: To investigate the type, length, and CG loci of HBV DNA CpG islands in HBsAg positive maternal C genotype and its relationship with intrauterine HBV transmission, so as to provide a new perspective for the study of intrauterine transmission of HBV. Methods: From June 2011 to July 2013, HBsAg-positive mothers and their newborns who delivered in the obstetrics and gynecology department of the Third People's Hospital of Taiyuan were collected. Epidemiological data were collected through face-to-face questionnaires and electronic medical records. Serum HBV markers and serum HBV DNA were detected by electrochemiluminescence and quantitative fluorescence PCR, respectively. Intrauterine transmission of HBV was determined by positive HBsAg and/or HBV DNA in femoral venous blood before injection of HBV vaccine/Hepatitis B immunoglobulin within 24 h of birth. A total of 22 mothers and their newborns with HBV DNA load ≥106 IU/ml in intrauterine transmission were selected as the intrauterine transmission group, and 22 mothers with HBV DNA load ≥106 IU/ml without intrauterine transmission were chosen as the control group by random seed method. The distribution prediction of CpG islands of HBV DNA in 39 mothers with genotype C by HBV DNA sequencing was analyzed. Results: Among 39 mothers with HBV C genotype, 19 were in the intrauterine transmission group, and 20 were in the control group. The HBV DNA of 39 patients with genotype C traditional CpG island Ⅱ and Ⅲ, while the control group had traditional CpG island Ⅰ and novel CpG island Ⅳ and Ⅴ. The length of CpG island Ⅱ and Ⅲ and the number of CG loci of CpG island Ⅱ in the intrauterine transmission group differed from those in the control group (P<0.05). The CpG island Ⅱ length ≥518 bp and the number of CG loci ≥40 in the intrauterine transmission group (11/19) were significantly higher than those in the control group (2/20) (P<0.05). The length of CpG island Ⅱ and the number of CG loci in the X gene promoter region (Xp region) were higher than those in the control group (P<0.05). In the HBV intrauterine transmission group, most of maternal (12/19) HBV DNA CpG island Ⅱ completely covered the Xp region, which was significantly higher than that in the control group (5/20), and the number of HBV DNA Xp region CG loci was higher than that in the control group (P<0.05). Conclusions: The distribution of maternal C genotype HBV DNA CpG islands is related to intrauterine transmission. The length of CpG island Ⅱ and the number of CG sites may increase the risk of intrauterine transmission of HBV.

Effect of activation of Toll-like receptor signaling pathway of peripheral blood mononuclear cell in recombinant hepatitis B surface antigen immune response / 中华流行病学杂志

Objective: To explore the effect and mechanism of activation of peripheral blood mononuclear cell (PBMC) Toll-like receptor (TLR3) signaling pathway in recombinant HBsAg (rHBsAg) immune response. Methods: White blood cells were collected from peripheral blood of 13 healthy donors in the preparation of blood products. PBMC was isolated and treated with Poly I:C (Poly I:C group) and PBS (control group) respectively. 48 h later, some cells were collected and the expressions of TLR3 signaling pathway proteins were detected by flow cytometry. After activating (Poly I:C group)/inactivating (control group) TLR3 signaling pathway, rHBsAg was given to both groups for 72 h, and the proportions of DC, T, B cells and their subsets in PBMC were detected by flow cytometry. Paired t-test, paired samples wilcoxon signed-rank test and canonical correlation analyses were used for statistical analysis. Results: The percentage of TLR3 protein-positive cells (19.21%) and protein expression (8 983.95), NF-κB protein expression (26 193.13), the percentage of pNF-κB protein-positive cells (13.73%) and its proportion in NF-κB (16.03%), and the percentage of pIRF3 protein-positive cells (12.64%) and its proportion in IRF3 (21.80%) in Poly I:C group were higher than those in control group (11.54%, 8 086.00, 22 340.66, 8.72%, 9.71%, 9.57%, 19.12%) (P<0.05), and the percentage of TRIF protein-positive cells (89.75%) and protein expression (304 219.54) were higher in Poly I:C group than in the control group (89.64%, 288 149.72) (P>0.05). After PBMC stimulation by rHBsAg, the proportions of mDC (2.90%), pDC (1.80%), B cell (5.31%) and plasma cell (67.71%) in Poly I:C group were significantly higher than those in the control group (1.83%, 0.81%, 4.23%, 58.82%) (P<0.05). Results of canonical correlation analysis showed that the expression of TLR3 protein was positively correlated with the proportions of plasma cells, the expression of pIRF3 protein was positively correlated with the proportions of plasma cells and mDC, and the percentage of pNF-κB protein-positive cells and the percentage of pIRF3 protein-positive cells were positively correlated with the proportion of CD4+T cells. Conclusions: Poly I:C can activate TLR3/TRIF/NF-κB and TLR3/TRIF/IRF3 signaling pathway, promote the function of downstream signaling molecules, and then promote the maturation of DC, induce the immune responses of CD4+T cell, and promote the maturation and activation of B cells and the immune response of rHBsAg.

Analysis on influencing factors of HBV intrauterine transmission based on integration of decision tree model and logistic regression model / 中华流行病学杂志

Objective: To investigate the influencing factors of HBV intrauterine transmission and their interaction effects by integrating logistic regression model and Chi-squared automatic interaction detector (CHAID) decision tree model. Methods: A total of 689 pairs of HBsAg-positive mothers and their neonates in the obstetrics department of the Third People's Hospital of Taiyuan from 2007 to 2013 were enrolled, and the basic information of mothers and their neonates were obtained by questionnaire survey and medical record review, such as the general demographic characteristics, gestational week and delivery mode. HBV DNA and HBV serological markers of the mothers and newborns were detected by fluorescence quantitative PCR and electrochemiluminescence immunoassay respectively. The CHAID decision tree model and unconditional logistic regression analysis were used to explore the factors influencing HBV intrauterine transmission in neonates of HBsAg-positive mothers. Results: Among the 689 neonates, the incidence of HBV intrauterine transmission was 11.47% (79/689). After adjusted for confounding factors, the first and second logistic multivariate analysis showed that cesarean delivery was a protective factor for HBV intrauterine transmission (OR=0.25, 95%CI: 0.14-0.43; OR=0.27, 95%CI: 0.15-0.46); both models indicated that maternal HBeAg positivity and HBV DNA load ≥2×105 IU/ml before delivery were risk factors of HBV intrauterine transmission (OR=3.89, 95%CI: 2.32-6.51; OR=3.48, 95%CI: 2.12-5.71), respectively. The CHAID decision tree model screened three significant factors influencing HBV intrauterine transmission, the most significant one was maternal HBeAg status, followed by delivery mode and maternal HBV DNA load. There were interactions between maternal HBeAg status and delivery modes, as well as delivery mode and maternal HBV DNA load before delivery. The rate of HBV intrauterine transmission in newborns of HBeAg-positive mothers by vaginal delivery increased from 19.08% to 29.37%; among HBeAg-positive mothers with HBV DNA ≥2×105 IU/ml, the rate of HBV intrauterine transmission increased to 33.33% in the newborns by vaginal delivery. Conclusions: Maternal HBeAg positivity,maternal HBV DNA ≥2×105 IU/ml and vaginal delivery could be risk factors for HBV intrauterine transmission in newborns. Interaction effects were found between maternal HBeAg positivity and vaginal delivery, as well as vaginal delivery and high maternal HBV DNA load. Logistic regression model and the CHAID decision tree model can be used in conjunction to identify the high-risk populations and develop preventive strategies accurately.

Comparison of effects of conservative treatment and percutaneous kyphoplasty on paravertebral muscle degeneration in patients with osteoporotic compression vertebral fractures / 中华骨科杂志

Objective:To compare effects of conservative treatment and percutaneous kyphoplasty on paravertebral muscle degeneration in patients with osteoporotic compression vertebral fractures.Methods:A retrospective case control analysis was conducted on 286 cases of osteoporotic compression vertebral fractures from January 2017 to December 2019. There were 54 males and 232 females, with a mean age of 67.7 (range, 52-90 years). According to the treatment, the patients were divided in to conservative treatment group (134 patients) and percutaneous kyphoplasty treatment group (152 patients). The pre-operation and post-operation of paravertebral muscle cross-sectional area (CSA) and fatty infiltration (FI% ) , bed rest time, visual analogue scale (VAS), Oswestry disability index (ODI), the sagittal view Cobb angle, and the anterior column height of fractured vertebra were compared between these two groups.Results:The two groups had no significant difference in CSA and FI% of paravertebral muscle in each plane of the intervertebral discs of the L 3-4、L 4-5 and L 5S 1. The CSA of multifidus in each plane of the intervertebral discs three months after operation were 6.56±1.26 cm 2, 6.87±1.31 cm 2, and 7.14±1.29 cm 2; the CSA of erector were 12.39±2.16 cm 2, 14.72±2.67 cm 2, and 16.45±3.09 cm 2; the CSA of psoas major were 7.05±1.52 cm 2, 8.12±1.75 cm 2, and 8.68±1.66 cm 2, which all were larger than those in conservative treatment group and showed significant difference between two groups ( P<0.05). However, the two groups had no significant difference in FI% of paravertebral muscle three months after operation. The CSA of multifidus in each plane of the intervertebral discs one year after operation were 6.43±1.23 cm 2, 6.62±1.42 cm 2, and 7.06±1.32 cm 2; the CSA of erector were 12.02±2.08 cm 2, 14.53±2.76 cm 2, and 16.39±2.84 cm 2; the CSA of psoas major were 6.98±1.47 cm 2, 8.01±1.59 cm 2, and 8.37±1.72 cm 2, which all were larger than those in conservative treatment group and showed significant difference between two groups ( P<0.05). The FI% of multifidus in each plane of the intervertebral discs one year after operation were 31.40%±5.84% , 32.54%±6.64% , and 33.26%±7.16% ; the FI% of erector were 22.64%±3.47% , 23.08%±3.72% , and 23.84%±3.99% ; the FI% of psoas major were 9.23%±2.20% , 9.72%±2.54% , and 10.98%±2.43% , which all were less than those in conservative treatment group and showed significant difference between two groups ( P<0.05). Two groups had significant difference in bed rest time as (9.21±2.52) d vs. (40.32±9.79) d ( t=37.79, P<0.001). The VAS, ODI score at the time of the first day after treatment and the last follow-up of the surgical treatment group were all significantly lower than those of conservative treatment group ( P<0.05). The operation could effectively improve the kyphosis deformity and reduce the loss the anterior column height of fractured vertebra compared with conservative treatment ( P<0.05). Conclusion:There exists paravertebral muscle degeneration of varying degrees during the course of the osteoporotic compression vertebral fractures. Compared to conservative treatment, percutaneous kyphoplasty treatment can not only significantly relieve pain in the short term, improve quality of patient's life, but also significantly delay the degeneration of paravertebral muscle.

Roles of transforming growth factor - β1 and heat shock protein 47 in progression of Schistosoma japonicum-induced hepatic fibrosis / 中国血吸虫病防治杂志

Objective To investigate the dynamic expression of transforming growth factor-β1 (TGF-β1) and heat shock protein 47 (HSP47) and explore their roles in the progression of hepatic fibrosis induced by Schistosoma japonicum infection. Methods Fifty female mice of the ICR strain were randomly divided into the infection group and the normal control group, of 25 mice in each group. Each mouse in the infection group was infected with 20 ± 1 cercariae of S. japonicum via the abdominal skin, while uninfected animals served as normal control. Five mice were sacrificed 4, 6, 8, 10 and 12 weeks post-infection and liver tissues were sampled. Serum HSP47 and TGF-β1 was determined using enzyme-linked immunosorbent assay (ELISA), and the pathological changes of liver specimens were observed with hematoxylin & eosin (HE) staining. In addition, the synthesis of alpha 1 chain of type I collagen (COL1A1) was measured using Masson staining, and the mRNA expression of TGF-β1, HSP47 and COL1A1 was determined using real-time fluorescent quantitative PCR (qPCR) assay. Results During the period of S. japonicum-induced hepatic fibrosis, the serum HSP47 and TGF-β1 levels and the mRNA expression of TGF - β1, HSP47 and COL1A1 gradually increased with the progression of hepatic fibrosis. The serum levels of HSP47 and TGF-β1 were (179.26 ± 29.87) pg/mL and (22.37 ± 5.21) ng/mL 6 weeks post-infection, respectively, which were significantly greater than those [(150.29 ± 34.91) pg/mL and (18.54 ± 7.78) ng/mL, respectively] in the normal control group (both P values < 0.05). In addition, the mRNA expression of HSP47, COL1A1 and TGF-β1 was (0.86 ± 0.04), (1.17 ± 0.06) and (0.64 ± 0.13) in mouse liver specimens, which was significantly higher than that (0.23 ± 0.03, 0.20 ± 0.02 and 0.38 ± 0.02) in the normal control group (all P values < 0.01). Conclusions The expression of TGF-β1 and HSP47 during the period of S. japonicum-induced hepatic fibrosis is consistent with the progression of the hepatic fibrosis, and exhibits the same tendency with type I collagen expression. HSP47 is a novel promising diagnosis marker and therapeutic target for S. japonicum-induced hepatic fibrosis.

Roles of transforming growth factor - β1 and heat shock protein 47 in progression of Schistosoma japonicum-induced hepatic fibrosis / 中国血吸虫病防治杂志

Objective To investigate the dynamic expression of transforming growth factor-β1 (TGF-β1) and heat shock protein 47 (HSP47) and explore their roles in the progression of hepatic fibrosis induced by Schistosoma japonicum infection. Methods Fifty female mice of the ICR strain were randomly divided into the infection group and the normal control group, of 25 mice in each group. Each mouse in the infection group was infected with 20 ± 1 cercariae of S. japonicum via the abdominal skin, while uninfected animals served as normal control. Five mice were sacrificed 4, 6, 8, 10 and 12 weeks post-infection and liver tissues were sampled. Serum HSP47 and TGF-β1 was determined using enzyme-linked immunosorbent assay (ELISA), and the pathological changes of liver specimens were observed with hematoxylin & eosin (HE) staining. In addition, the synthesis of alpha 1 chain of type I collagen (COL1A1) was measured using Masson staining, and the mRNA expression of TGF-β1, HSP47 and COL1A1 was determined using real-time fluorescent quantitative PCR (qPCR) assay. Results During the period of S. japonicum-induced hepatic fibrosis, the serum HSP47 and TGF-β1 levels and the mRNA expression of TGF - β1, HSP47 and COL1A1 gradually increased with the progression of hepatic fibrosis. The serum levels of HSP47 and TGF-β1 were (179.26 ± 29.87) pg/mL and (22.37 ± 5.21) ng/mL 6 weeks post-infection, respectively, which were significantly greater than those [(150.29 ± 34.91) pg/mL and (18.54 ± 7.78) ng/mL, respectively] in the normal control group (both P values < 0.05). In addition, the mRNA expression of HSP47, COL1A1 and TGF-β1 was (0.86 ± 0.04), (1.17 ± 0.06) and (0.64 ± 0.13) in mouse liver specimens, which was significantly higher than that (0.23 ± 0.03, 0.20 ± 0.02 and 0.38 ± 0.02) in the normal control group (all P values < 0.01). Conclusions The expression of TGF-β1 and HSP47 during the period of S. japonicum-induced hepatic fibrosis is consistent with the progression of the hepatic fibrosis, and exhibits the same tendency with type I collagen expression. HSP47 is a novel promising diagnosis marker and therapeutic target for S. japonicum-induced hepatic fibrosis.

Expressions of TOLL-like receptor 4 (TLR-4) and matrix metalloproteinase 9 (MMP-9)/Tissue inhibitor of metalloproteinase 1 (TIMP-1) in pulmonary blood vessels with chronic obstructive pulmonary diseases and their relationships with pulmonary vascular remodelling

SUMMARY OBJECTIVE: This study aims at investigating the expressions of TOLL-like receptor 4 (TLR-4) and matrix metalloproteinase 9 (MMP-9)/ tissue inhibitor of metalloproteinase 1 (TIMP-1) in pulmonary blood vessels with chronic obstructive pulmonary disease (COPD) and their relationships with pulmonary vascular remodelling (PVR). METHODS: 60 para-tumour tissues were divided into the COPD group and the control group (n=30); the inflammations, pulmonary artery wall area/total artery area (WA%), and wall thickness/vascular outer diameter (WT%) were compared. The expressions of TLR-4, MMP-9/TIMP-1, and PCNA in pulmonary vascular smooth muscle cells were detected, and their relationships with PVR were then analysed. RESULTS: The inflammations (1.6±0.8), WA% (44.0±6.4), and WT% (27.3±3.3) in the COPD group were higher than in the control group (0.3±0.5, 26.1±2.8, 15.6±1.8), and the expressions of TLR-4 (31.4±147) and MMP-9/TIMP-1 (2.2±2.6) were increased compared to the control group (4.7±4.5, 1.9±12). Correlation analysis: TLR-4 and MMP-9/TIMP-1 were positively correlated with the inflammations (r=0.18, P<0.01), WA% (r=0.68, P<0.01), and WT% (r=0.73, P<0.01), as well as positively correlated with the expression of PCNA (r=0.44, P<0.01); the upregulation of TLR-4 was positively correlated with the expressions of MMP-9 and TIMP-1. CONCLUSIONS: The upregulation of TLR-4 in the pulmonary arterial smooth muscle cells of COPD patients could promote the inflammations and the MMP-9 expression, thus causing abnormal degradation of extracellular matrix, so it played an important role in the process of PVR.

RESUMO OBJETIVO: Este estudo tem como objetivo investigar as expressões de TOLL-like receptor 4 (TLR-4) e metaloproteinase 9 da matriz (MMP-9)/inibidor de tecido da metaloproteinase 1 (TIMP-1) em vasos sanguíneos pulmonares com doença pulmonar obstrutiva crônica (DPOC) e suas relações com o remodelamento vascular pulmonar (PVR). MÉTODOS: Sessenta tecidos paratumorais foram divididos em grupo COPD e o grupo controle (n = 30). Foram comparadas as inflamações, área da parede da artéria pulmonar/área da artéria total (WA%) e espessura da parede/diâmetro externo vascular (WT%). As expressões de TLR-4, MMP-9/TIMP-1 e PCNA em células de músculo liso vascular pulmonar foram detectadas, e suas relações com PVR foram então analisadas. RESULTADOS: As inflamações (1,6 ± 0,8), WA% (44,0 ± 6,4) e WT% (27,3 ± 3,3) no grupo COPD foram maiores que no grupo controle (0,3 ± 0,5; 26,1 ± 2,8; 15,6 ± 1,8). E as expressões de TLR-4 (31,4 ± 14,7) e MMP-9/TIMP-1 (2,2 ± 2,6) foram aumentadas em relação ao grupo controle (4,7 ± 4,5, 1,9 ± 1,2). Na análise de correlação, TLR-4 e MMP-9/TIMP-1 foram positivamente correlacionadas com as inflamações (r = 0,18; P <0,01), WA% (r = 0,68; P <0,01) e WT% (r = 0,73; P <0,01), bem como correlacionadas positivamente com a expressão de PCNA (r = 0,44; P <0,01). A elevação da TLR-4 foi correlacionada positivamente com as expressões de MMP-9 e TIMP-1. CONCLUSÕES: A regulação positiva do TLR-4 nas células do músculo liso arterial pulmonar de pacientes com DPOC poderia promover as inflamações e a expressão de MMP-9, causando assim uma degradação anormal da matriz extracelular, por isso desempenhou um papel importante no processo de PVR.

Comparison of clinical efficacy of unilateral and bilateral short segment transpedicular fixation in treating thoracolumbar fractures combined with unilateral pedicle fractures / 中华创伤杂志

Objective To compare the clinical efficacy of unilateral and bilateral short segment fixation in treating thoracolumbar fractures combined with unilateral pedicle fractures.Methods A retrospective case control study was conducted on the clinical data of 43 patients with thoracolumbar fractures with unilateral pedicle fractures admitted from January 2012 to December 2016.There were 24 males and 19 females,with a mean age of 48 years (range,19-68 years).Fractured segments included T10 in 1 patient,T11 in 6,T12 in 11,L1 in 19,and L2 in 6.According to the fixation method,the patients were divided into unilateral transpedicular fixation group (unilateral group,15 patients) and bilateral transpedicular fixation group (bilateral group,28 patients).All patients were treated with fixation via injured vertebrae combined with short segment fixation.The operation time,intraoperative blood loss,X-ray frequency,fracture healing time,intemal fixation removal time,preoperative and postoperative anterior vertebral height ratio,sagittal Cobb angle recovery,spinal canal invasion rate,visual analogue score (VAS),Oswestry dysfunction index (ODI),Frankel classification of spinal cord injury,incidence of screw deviation,and spinal lateral angulation were compared between the two groups.Results All patients were followed up for 12-26 months,with an average of 18.5 months.The operation time in unilateral group and bilateral group were (82.3 ± 14.7) minutes and (120.9 ± 12.8) minutes,respectively.The intraoperative blood loss was (186.1 ± 20.4)ml in unilateral group and (231.2 ± 39.6) ml in bilateral group.The number of fluoroscopy was (6.6 ± 1.2) times in unilateral group and (13.3 ± 2.0) times in bilateral group respectively.The incidence of screw deviation was 13％ (2/15) in unilateral group and 46％ (26/56) in bilateral group (P ＜ 0.05).There were no significant differences between the two groups in anterior vertebral height ratio,sagittal Cobb angle,spinal canal invasion rate,VAS,ODI,Frankel grade and spinal lateral angulation (P ＞ 0.05).Conclusions For thoracolumbar fractures combined with unilateral pedicle fractures,unilateral and bilateral transpedicular short segment fixation has similar reduction and fixation effects.Unilateral transpedicular short segment fixation has the advantages of shorter operation time,less bleeding,fewer fluoroscopy times,and lower screw deviation rate.

Septation of the Intrapericardial Arterial Trunks in the Early Human Embryonic Heart / 中华医学杂志(英文版)

<p><b>Background</b>Outflow tract (OFT) septation defects are a common cause of congenital heart disease. Numerous studies have focused on the septation mechanism of the OFT, but have reported inconsistent conclusions. This study, therefore, aimed to investigate the septation of the aortic sac and the OFT in the early embryonic human heart.</p><p><b>Methods</b>Serial sections of 27 human embryonic hearts from Carnegie stage (CS) 10 to CS19 were immunohistochemically stained with antibodies against α-smooth muscle actin (α-SMA) and myosin heavy chain.</p><p><b>Results</b>At CS10-CS11, the OFT wall was an exclusively myocardial structure that was continuous with the aortic sac at the margin of the pericardial cavity. From CS13 onward, the OFT was divided into nonmyocardial and myocardial portions. The cushion formed gradually, and its distal border with the OFT myocardium was consistently maintained. The aortic sac between the fourth and sixth aortic arch arteries was degenerated. At CS16, the α-SMA-positive aortopulmonary septum formed and fused with the two OFT cushions, thus septating the nonmyocardial portion of the OFT into two arteries. At this stage, the cushions were not fused. At CS19, the bilateral cushions were fused to septate the myocardial portion of the OFT.</p><p><b>Conclusions</b>Data suggest that the OFT cushion is formed before the aortopulmonary septum is formed. Thus, the OFT cushion is not derived from the aortopulmonary septum. In addition, the nonmyocardial part of the OFT is septated into the aorta and pulmonary trunk by the aortopulmonary septum, while the main part of the cushion fuses and septates the myocardial portion of the OFT.</p>

Application of fuzzy comprehensive evaluation model in configuration management of medical equipment / 中国医学装备

Objective: To explore the optimal configuration methods of medical equipment of hospital so as to increase the scientific, rationality, fairness and effectiveness of decisions for equipment configuration of hospital. Methods:The applications of ultrasound imaging equipment configuration of two departments were used as example. Through a series of steps, included establishing the hierarchical structure of model, constructing the judgment matrix, consistency check, determining the weight matrix, determining the evaluation set and fuzzy evaluation matrix, and calculating the evaluation vector quantity and so on, the fuzzy comprehensive evaluation model was performed and applied in the evaluation of equipment configuration. Results: After evaluation, for the ultrasound imaging equipment configuration of department 1, the ratio of experts who thought the introduction of this equipment was very good was 82.92%, and the ratio of experts who thought it was better was 12.36%, and the ratio of experts who though it was normal was 4.72%. While for department 2, the ration of experts who though it was normal or worst was more than 50%. Conclusion:The fuzzy comprehensive evaluation model is an effective method for verifying equipment configuration. It can provide effective and quantitative assessment results for decision makers of hospital and provide reference for optimal configuration of medical equipment of hospital.

Comparison of commercial HIV-1 viral load tests by using proficiency test results in China,2013-2015 / 中华流行病学杂志

Objective To compare the bio-equivalence among commercial HIV-1 viral load tests,including EasyQ HIV-1 v2.0 (EasyQ) from bioMerieux NucliSens of France;VERSANT HIV-1 RNA 3.0 assay (bDNA) from Siemens Healthcare Diagnostics of USA;COBAS AmpliPrep/COBAS TaqMan HIV-1 test (Taqman) from Roche Molecular Diagnosis of USA;Abbott Real Time HIV-1 Kit (M2000) from Abbott Molecular of USA and two domestic HIV-1 viral load test kits (domestic kit) from DaAn Gene Company of Sun Yat-Sen University and Liaoning Bio-Pharmaceutical company of Northeast pharmaceutical group,by using proficiency test results in China from 2013 to 2015.Methods A total of 2 954 proficiency test results,obtained from 22 positive samples of 6 proficiency tests in 155 laboratories conducted by China CDC were analyzed during 2013-2015.The results from each sample were first logarithmic transformed and then grouped according to the method used,the mean value of logarithmic results was calculated.Subsequently,22 clusters of mean values were analyzed by Bland-Altman analysis for the consistency,and linear regression analysis for the interdependency.Results The results indicated that,by taking Taqman as the reference,EasyQ,M2000,bDNA and domestic kit had good consistency (90％-100％) and interdependency.Conclusion All the viral load tests were bio-equivalent.Moreover,according to the conversion formula derived from domestic proficiency test results,all the viral load results could be converted,which is critical for epidemiological analysis.

Comparison of commercial HIV-1 viral load tests by using proficiency test results in China,2013-2015 / 中华流行病学杂志

Objective To compare the bio-equivalence among commercial HIV-1 viral load tests,including EasyQ HIV-1 v2.0 (EasyQ) from bioMerieux NucliSens of France;VERSANT HIV-1 RNA 3.0 assay (bDNA) from Siemens Healthcare Diagnostics of USA;COBAS AmpliPrep/COBAS TaqMan HIV-1 test (Taqman) from Roche Molecular Diagnosis of USA;Abbott Real Time HIV-1 Kit (M2000) from Abbott Molecular of USA and two domestic HIV-1 viral load test kits (domestic kit) from DaAn Gene Company of Sun Yat-Sen University and Liaoning Bio-Pharmaceutical company of Northeast pharmaceutical group,by using proficiency test results in China from 2013 to 2015.Methods A total of 2 954 proficiency test results,obtained from 22 positive samples of 6 proficiency tests in 155 laboratories conducted by China CDC were analyzed during 2013-2015.The results from each sample were first logarithmic transformed and then grouped according to the method used,the mean value of logarithmic results was calculated.Subsequently,22 clusters of mean values were analyzed by Bland-Altman analysis for the consistency,and linear regression analysis for the interdependency.Results The results indicated that,by taking Taqman as the reference,EasyQ,M2000,bDNA and domestic kit had good consistency (90％-100％) and interdependency.Conclusion All the viral load tests were bio-equivalent.Moreover,according to the conversion formula derived from domestic proficiency test results,all the viral load results could be converted,which is critical for epidemiological analysis.

Posterior enlargement of spinal canal for the treatment of multi-segmental cervical diseases without cervical lordosis / 中华骨科杂志

Objective To evaluate the feasibility and efficacy of posterior enlargement of spinal canal for the treatment of multi-segmental cervical diseases without cervical lordosis.Methods From January 2013 to June 2017,a retrospective study was conducted with 21 patients of multi-segmental cervical diseases accompanied cervical lordosis loss,and the complete followup data was obtained.There were 14 males and 7 females,with an average age of 53.9±7.3 years (range,42-65 years).There were 14 multi-segmental cervical spondylotic myelopathy,5 ossification of posterior longitudinal ligament,and 2 congenital cervical stenosis included in this study.The cervical lordotic angle and cervical curvature index were measured preoperatively and 1 year postoperatively.To access the enlargement of spinal canal and spinal cord,the anteroposterior diameter and cross section area of spinal canal or spinal cord were measured on MRI preoperatively and 1 year postoperatively.The Japanese Orthopaedic Association Scores (JOA) was applied to evaluate the neurological function at preoperation and postoperation.Visual Analogue Scales (VAS) was applied to evaluate the pain degree at preoperation and postoperation.Frankel classification was used to assess the severity of spinal cord injury at preoperation and postoperation.Results The follow-up time was 12-26 months,with an average of 16.4 months.The cervical lordosis angle was 3.1°±2.3° preoperatively,and 4.2°±1.6° 1 year postoperatively with a significant difference.The cervical curvature index was 4.4％ ± 1.7 ％ preoperatively and 5.0％ ± 1.5 ％ 1 year postoperatively with no statistically difference.Except for C7T1 level,the preoperative anteroposterior diameter and cross section area of spinal canal at C2.3,C3.4,C4.5,C5.6,and C6.7 level were lower than that at 1 year after operation with a significant difference.Except for C2,3 and C7T1 and level,the preoperative anteroposterior diameter and cross section area of spinal cord at C3,4,C4,5,C5,6,and C6,7 level were significantly lower than that at 1 year after operation.The average JOA score preoperatively was 8.9±1.7.The average JOA score at 3 months postoperatively was 13.1±2.0,which was significantly higher than that preoperatively.At 3 months postoperatively,the average improvement rate was 52.0％,and the superior rate was 52.3 ％.At 1 year postoperatively,the average JOA score was 13.3±2.1,which improved significantly from that preoperatively.The average improvement rate was 54.3 ％,and the superior rate was 61.9％.The VAS score at preoperatively was 3.0±2.4,and which was 2.7± 1.7 at 1 year postoperatively with no significant differences.At pre-operation,the level of Frankel classification was C level in one (4.8％) case,D level in 8 (38.1％) cases and E level in 12 (57.1％) cases.At 1 year postoperatively,the level of Frankel classification was C level in one (4.8％) case,D level in 6 (28.6％) cases and E level in 14 (66.7％) cases,compared with that at preoperatively,there was no statistically significant difference.One patients suffered from neurologic deterioration at 1 year after surgery and recovered after anterior cervical surgery.No other serious complications were occurred.Conclusion For the patients with multi-segmental cervical diseases accompanied cervical lordosis loss,effective spinal decompression by cervical posterior laminoplasty was feasible,and a good clinical efficacy was achieved.

Applied anatomic study of narrow pedicle in thoracolumbar spine of adults / 中华骨科杂志

Objective To explore the demographic factors and the risk of the pedicle screw insertion of the narrow pedi?cles. Methods Thoracolumbar spine thin?section CT image data of 312 adults from September 2014 to September 2015 were ana?lyzed. The pedicle width,medial and lateral cortical thickness, spongy bone thickness, spongy bone thickness/cortical thickness, e angle and screw path length of each pedicle were measured. The incidence and the distribution characteristics of the narrow pedi?cle were analyzed. Anatomic parameters and age, gender and stature were compared between the narrow pedicle group and non?narrow pedicle. The risk of the pedicle screw insertion of the narrow pedicle was assessed. Results Among the 3 081 pedicles, 74 narrow pedicles were determined as their pedicles width were less than 5 mm, and the proportion of narrow pedicle was 2.40%. Among the 312 subjects, 26 subjects were found having narrow pedicles, and the proportion of individuals with narrow pedicles in the population was 8.33% (26/312). The incidences of narrow pedicle in thoracolumbar spine were T10 0.32%, T11 0.32%, T12 0.98%, L1 7.54%, L2 2.92%. The spongy bone thickness, spongy bone thickness/cortical thickness of narrow pedicle were lower than non?narrow pedicle. However, there were no significant differences of medial and lateral cortical thickness, e angle and screw path length between the narrow pedicle and non?narrow pedicle. Difference of the mean age between the two subjects groups had no statistical significance. The percentage of female in narrow pedicle subjects group was 84.6%(22/26), which was higher than that in non?narrow pedicle subjects group (49.7%, 142/286). The mean stature of the male and female of stenosis pedicle group subjects were 163.8±1.3 cm and 152.5±4.3 cm, which were shorter than those of non?narrow subjects pedicle group (169.5±5.6 cm, 160.1±6.6 cm). The percentage of the cortical bone breakthrough by the pedicle screws of narrow pedicle group was (84.6%, 27/32), which was higher than that of non?narrow pedicle group (14.7%, 33/224). Conclusion L1 is the most common segment of thoracolumbar spine that narrow pedicle exist, which is the result of reduction of the spongy bone thickness. Narrow pedicle mostly appears in short stature female. There is high risk of cortical bone breakthrough by insertion of the posterior pedicle screws in the narrow pedicle.

Inclusion Bodies are Formed in SFTSV-infected Human Macrophages / 病毒学报

The severe fever with thrombocytopenia syndrome virus (SFTSV) is a new member in the genus Phlebovirus of the family Bunyaviridae identified in China. The SFTSV is also the causative pathogen of an emerging infectious disease: severe fever with thrombocytopenia syndrome. Using immunofluorescent staining and confocal microscopy, the intracellular distribution of nucleocapsid protein (NP) in SFTSV-infected THP-1 cells was investigated with serial doses of SFTSV at different times after infection. Transmission electron microscopy was used to observe the ultrafine intracellular structure of SFTSV-infected THP-1 cells at different times after infection. SFTSV NP could form intracellular inclusion bodies in infected THP-1 cells. The association between NP-formed inclusion bodies and virus production was analyzed: the size of the inclusion body formed 3 days after infection was correlated with the viral load in supernatants collected 7 days after infection. These findings suggest that the inclusion bodies formed in SFTSV-infected THP-1 cells could be where the SFTSV uses host-cell proteins and intracellular organelles to produce new viral particles.

Infection of the Severe Fever with Thrombocytopenia Syndrome Virus in Balb/C Mice and Hamsters / 病毒学报

The severe fever with thrombocytopenia syndrome virus (SFTSV) is the causative pathogen of an emerging infectious disease severe fever with thrombocytopenia syndrome and a new member in the genus Phlebovirus of family Bunyaviridae. Immune responses and pathological lesions in SFTSV-infected Balb/C mice and hamsters were evaluated by inoculation of SFTSV at 105 TCID50 or 103 TCID50 per animal through four different routes of infection, including intravenous, intramuscular, intraperitoneal, and intracerebral injections. The vehicle control groups were also included. At different time points after the inoculation blood and plasma samples were collected. Blood cell counts, blood viral RNA copies, and plasma antibodies were detected by automatic blood cell counters, real-time PCR, and luminex assays, respectively. At two weeks post inoculation, the animals were sacrificed. Tissues including heart, liver, spleen, lung, kidney, intestine, muscle, and brain, were collected for pathological analyses. Results showed that the SFTSV could infect Balb/C mice and hamsters with SFTSV-specific immunoglobulin (Ig) M and IgG antibodies detected in plasma samples on day 7 post inoculation. The SFTSV-specific IgM levels peaked on day 7 post inoculation and then decreased, whereas the SFTSV-specific IgG levels started to increase on day 7 and then peaked on day 14 post inoculation. Pathological analyses indicated significant pathological lesions in liver and kidney tissues. In conclusion, SFTSV could can infect different strains of rodent animals and cause similar immunological and pathological responses.

Association of CRP and D-Dimer with secondary pulmonary hypertension in patients with AECOPD / 实用医学杂志

Objective To explore the clinical significance of C-reaction protein ( CRP ) and D-Dimer levels in patients with acute exacerbation of chronic obstructive pulmonary disease ( AECOPD ) complicated by pulmonary hypertension (PH) and their association with HP. Methods Arterial blood gas and levels of CRP and D-Dimer were detected in 150 patients with AECOPD. Results Levels of CRP, D-Dimer, and PaCO2 were higher but PO2 level was lower in patients with moderate to severe PH than those with mild PH and the control subjects. Levels of CRP and D-Dimer were higher in patients with mild PH than the control subjects. Levels of CRP, D-Dimer, and PCO2 levels had a linear relationship with PASP, while PO2 was negatively correlated with PASP. Levels of CRP and D-Dimer were positively related with PCO2, while were negatively correlated with PO2. Conclusions Levels of CRP and D-Dimer can be used as an indicator for estimating the severity of pulmonary hypertension in patients with AECOPD.