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OBJECTIVE@#To investigate whether Salvianolic acid A (SAA) can restore cartilage endplate cell degeneration of intervertebral discs and to identify the mechanism via regulation of micro-RNA.@*METHODS@#Cartilage endplate cells were isolated from lumbar intervertebral disc surgical samples and were treated with serum containing a series of concentrations of SAA (2, 5, and 10 ?M) for 24, 48, and 72 h to identify a proper dose and treatment time of SAA. The effect SAA on interlenkin-1β (IL-1β)-induced extracellular matrix degradation of cartilage endplate cells were analyzed by Alcian blue staining and assessment of the expression levels of ADAMTS-5, MMP3 and Col2a1. Further, the potential target miRNAs were preliminarily screened by micro-RNA sequencing combining qRT-PCR and Western blot, and then, the miRNAs mimics and inhibitors were used to verify the regulatory effect of SAA on potential target miRNAs.@*RESULTS@#The 10 μM SAA treatment for 48 h significantly enhanced the viability of cartilage endplate cells, and increased Col2a1 expression and glycosaminoglycan accumulation that were repressed by IL-1β, and reduced the effect of IL-1β on ADAMTS-5, and MMP3. Screening analysis based on micro-RNA sequencing and Venny analysis identified the downstream micro-RNAs, including miR-940 and miR-576-5p. Then, the miR-940-mimic or miR-576-5p-mimic were transfected into CEPCs. Compared with the SAA group, the expression of ADAMTS-5 and MMP3 increased significantly and the expression of COL2A1 obviously decreased after overexpression of miR-940 or miR-576-5p in CEPCs.@*CONCLUSION@#Salvianolic acid A attenuated the IL-1β-induced extracellular matrix degradation of cartilage endplate cells by targeting regulate the miR-940 and the miR-576-5p.
Subject(s)
Humans , Apoptosis , Cartilage/metabolism , Chondrocytes/metabolism , Interleukin-1beta/metabolism , Matrix Metalloproteinase 3/metabolism , MicroRNAs/metabolismABSTRACT
ObjectiveTo examine the changes in the immune functions of CD8+ T cells in the spleen of mice following Echinococcus multilocularis infections at various doses and at different time points. MethodsThe E. multilocularis protoscoleces were collected, and E. multilocularis infection was modeled in mice via the hepatic portal vein at doses of 50 (low-dose), 500 (medium-dose) and 2 000 protoscoleces (high-dose), while physiological saline served as controls. Mouse spleen was isolated 2 (earlystage), 12 (middle-stage) and 24 weeks post-infection (late-stage), and spleen lymphocytes were harvested. The phenotype of memory CD8+ T cells and 2B4 expression were quantified in the mouse spleen, and the secretion of interferon (IFN)-γ, tumor necrosis factor (TNF)-α, interleukin (IL)-17A and IL-10 was measured. Results A central-memory phenotype was predominant in the CD8+ T cells in the spleen of mice at the early stage of high-dose protoscolece infections, and the proportion of central-memory CD8+ T cells was significantly greater in the high-dose group than in the control group (35.50% ± 2.00% vs. 25.90% ± 2.46%, P < 0.01), while a effector- memory phenotype was predominant in the CD8+ T cells in the spleen of mice at the late stage of medium- and high-dose protoscolece infections, and the proportions of effector-memory CD8+ T cells were significantly greater in the medium- (25.70% ± 4.12%) and high-dose group (28.40% ± 4.12%) than in the control group (10.50% ± 6.45%) (P < 0.05). The proportions of the central-memory CD8+ T cells were significantly higher in the high-dose group than at middle and late stages than at the early stage (P < 0.01), and the proportion of effector-memory CD8+ T cells was significantly greater in the high-dose group at the late stage than at early and middle stages (P < 0.05). The secretion of IFN-γ and IL-17A by spleen CD8+ T cells was elevated in the low- and medium-dose groups at the early stage of infection, and high-dose protoscolece infection promoted the secretion of IFN-γ and TNF-α by spleen CD8+ T cells; however, the levels of IFN-γ and TNF-α were significantly lower at the late stage than at the early and middle stages (P < 0.05). In addition, high 2B4 expression was detected in spleen CD8+ T cells in the middle- and high-dose groups at the late stage of infection, and the 2B4 expression was significantly higher in the medium(4.73% ± 1.56%) and high-dose groups (4.94% ± 1.90%) than in the low-dose group (2.49% ± 0.58%) and the control group (2.92% ± 0.60%) (P < 0.05). Conclusions E. multilocularis may be killed and eliminated through the host immune responses at the middle and late stages of low- and medium-dose protoscolece infections, while high-dose protoscolece infections may trigger the upregulation of 2B4 expression in mouse spleen CD8+ T cells at the late stage, which leads to immune exhaustion and the resultant chronic infections.
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BACKGROUND: Animal models are critical to study the mechanism, prevention and treatment of intervertebral disc degeneration (IDD). Therefore, constructing an ideal animal model of IDD is the key to further study IDD. OBJECTIVE: To review the selection and construction methods of the IDD model, so as to select and construct an ideal animal model of IDD. METHODS: A retrieval of CNKI, WanFang, VIP, SinoMed and PubMed databases was performed for the articles published before December 2016. The keywords were "intervertebral disc degeneration, animal model" in English and Chinese, respectively. All the articles were selected from the authoritative magazines, and finally 56 eligible articles were included. RESULTS AND CONCLUSION: There are many kinds of animals used for constructing the IDD model, including small and large animals. The former has a small volume of intervertebral disc that is beneficial for nutrient and metabolite transport,so it can be used for long-term in vitro culture.The latter has a large volume of intervertebral disc,which is appropriate for biomechanical study.The animal models of IDD include in vivo and in vitro models:the in vivo models include the changed biomechanics,destroyed physical structure,spontaneous and systemic disease models;the in vitro models include in vitro cellular and organ models.However,there is still a lack of an ideal animal model that can fully simulate human IDD. Noticeably, similarity, comparability, economy, feasibility, reliability and controllability should be considered.
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<p><b>OBJECTIVE</b>To evaluate the stability of teaching robot of rotation-traction manipulation.</p><p><b>METHODS</b>Operators were required to get the hang of rotation-traction manipulation and had clinical experience for over 5 years. The examination and data processing of the ten operators in our research were collected by the teaching robot of rotation-traction manipulation. Traction, pulling force, maximum force, pulling time, rotational amplitude and pitch range were recorded and compared for five times(G1, G2, G3, G4 and G5). The qualification rates were analyzed to evaluate the stability of teaching robot of rotation-traction manipulation.</p><p><b>RESULTS</b>Nonconforming items were found in G1 and G2, for instance, pulling force(=0.074), maximum force(=0.264) and rotational amplitude (=0.531). There was no statistically difference. None nonconforming item was found in G3, G4 and G5. All data were processed by SPSS and One-way ANOVA was used to analysis. Pulling force was found statistically different in G1, compared with G4 and G5(=0.015,=0.006). Maximum force was found statistically different in G1, compared with G4 and G5 (=0.021,=0.012). None differences were found in other comparisons (>0.05).</p><p><b>CONCLUSIONS</b>The teaching robot of rotation-traction manipulation used in our research could provide objective and quantitative indices and was considered to be an effective tool of assessing the rotation-traction manipulation.</p>
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<p><b>OBJECTIVE</b>To study the effect of Bushen Huoxue decoction on calcification of cartilage endplate in lumbar vertebrae.</p><p><b>METHODS</b>Six healthy male gerbils with 2-month-old were selected as normal control group, and 24 7-month-old healthy male gerbils were fed to 12-month-old to establish the aged gerbil model. Thirty gerbils were randomly divided into five groups as follow: the normal control group (=6), model group (=6, normal saline 4 ml/kg, intragastric 30 d), Bushen Huoxue low dose group(=6, 1.9× 10⁻ ³ ml/g given Bushen Huoxue recipe orally, 30 d), Bushen Huoxue middle dose group(=6, 3.8× 10⁻ ³ ml/g given Bushen Huoxue recipe orally, 30 d), Bushen Huoxue high dose group(=6, 7.6× 10⁻ ³ ml/g given Bushen Huoxue recipe orally, 30 d), the intervention group administered for 1.36 g from 7-month-old age, 30 d. The animals were sacrificed at the age of 2 months in the normal control group and 12 months of age in the other groups. The morphology of the lumbar vertebral cartilage endplate, the area of vascular bud, the ratio of non-calcified/calcified layer were analysis by HE chromosome visual method. The expression of type X collagen and BMPs in cartilage endplates were detected by rabbit monoclonal immunohistochemical staining.</p><p><b>RESULTS</b>The relative area of the vascular buds cartilage endplate measurements showed that compared with the model group, middle dose group and normal control group increased (<0.05), high and low dose groups all had different degrees of increase, but no statistical significance(>0.05). The ratio of cartilage endplate thickness of non-calcified/calcified showed that compared with the model group, Bushen Huoxue middle dose, normal control group increased, with statistical significance(<0.05), and high and low dose groups all had different degrees of increase, but there were no statistical significance(>0.05). Compared with the model group, the expression of type X collagen in the cartilage endplate of the normal group, the Bushen Huoxue low, middle and high dose groups decreased, and had statistical significance(<0.01); compared with the model group, the expression of BMPs in the normal group, Bushen Huoxue middle dose group increased, with statistically significant(<0.01), while the high and low dose groups increased in different degrees, but there was no statistical significance(>0.05).</p><p><b>CONCLUSIONS</b>Bushen Huoxue prescription can delay the calcification of cartilage endplate in the process of aging, suggesting that it can be used as a preventive medicine for early disc degeneration.</p>
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<p><b>OBJECTIVE</b>To study the efficacy and safety of voluntary rehabilitation exercise in the treatment of knee osteoarthritis, based on the traditional Chinese medicine theory of "treating muscle for bone".</p><p><b>METHODS</b>Ninety participants with early knee osteoarthritis were randomly divided into experimental group (=45) and control group (=45). Patients in experimental group were treated with voluntary rehabilitation exercise combined with isometric extension of quadriceps femoris. Patients in control group were treated with apparatus training combined with isometric extension of quadriceps femoris. The treatment course lasted for two weeks. Visual analogue scale (VAS), Lysholm score and total therapeutic effect were evaluated before and after treatments.</p><p><b>RESULTS</b>After two weeks of treatment, cure-remarkable-effective rate in experimental group (86.67%) was higher than that in control group (71.11%). The VAS scores and Lysholm scores were significantly improved in both two groups (<0.05). The results were significantly better in experimental group those in control group (<0.05). There were no serious adverse events.</p><p><b>CONCLUSIONS</b>Voluntary rehabilitation exercise combined with isometric extension of quadriceps femoris was effective and safe in the treatment of early knee osteoarthritis.</p>
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Objective To identify the differential proteomic expressions between the liver tissues of male and female mice, and investigate the mechanisms underlying gender differences in liver diseases. Methods Two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) and matrix-assisted laser desorption/ionization time of flight mass spectrometry ( MALDI-TOF-MS) were used to identify the differentially expressed proteins in the liver tissues of male and female C57BL/6J mice. The differentially expressed proteins were validated by Western blot and further analyzed by bioinformatics, including Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG). Results Among the auto-detected 1767 protein spots by 2D-DIGE, 325 protein spots were differentially expressed (|ratio|≥1. 5, P< 0. 05) between the liver tissues of male and female mice, in which 78 spots were randomly selected for MALDI-TOF-MS identification and finally 48 distinct proteins were obtained. Compared with females, 14 and 34 proteins were up-or down-regulated in males, respectively. Among them, 6 differentially expressed proteins were validated by Western blot which confirmed the reliability of 2D-DIGE results. GO analysis showed that the differentially expressed proteins in the liver tissues of male and female mice are associated to various cellular component, molecular function and biological process. 6 pathways were significantly different between the liver tissues of males and females depending on KEGG analysis. Conclusions The proteomic data and related analysis of the liver tissues of C57BL/6J mice offer crucial clues for elucidating the underlying mechanisms of different gender effects on liver diseases.
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Objective To identify the differential proteomic expressions between the liver tissues of male and female mice, and investigate the mechanisms underlying gender differences in liver diseases. Methods Two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) and matrix-assisted laser desorption/ionization time of flight mass spectrometry ( MALDI-TOF-MS) were used to identify the differentially expressed proteins in the liver tissues of male and female C57BL/6J mice. The differentially expressed proteins were validated by Western blot and further analyzed by bioinformatics, including Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG). Results Among the auto-detected 1767 protein spots by 2D-DIGE, 325 protein spots were differentially expressed (|ratio|≥1. 5, P< 0. 05) between the liver tissues of male and female mice, in which 78 spots were randomly selected for MALDI-TOF-MS identification and finally 48 distinct proteins were obtained. Compared with females, 14 and 34 proteins were up-or down-regulated in males, respectively. Among them, 6 differentially expressed proteins were validated by Western blot which confirmed the reliability of 2D-DIGE results. GO analysis showed that the differentially expressed proteins in the liver tissues of male and female mice are associated to various cellular component, molecular function and biological process. 6 pathways were significantly different between the liver tissues of males and females depending on KEGG analysis. Conclusions The proteomic data and related analysis of the liver tissues of C57BL/6J mice offer crucial clues for elucidating the underlying mechanisms of different gender effects on liver diseases.
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Objective To observe the effect of electrical stimulation combined with bladder function training on autonomic bladder pa-tients after S2-4spinal cord injury or cauda equina and pelvic nerve injury.Methods From January,2016 to August,2017,76 patients with au-tonomic bladder after spinal cord injury were divided into control group(n=29)and observation groups(n=47)in real world study.The con-trol group received routine bladder function training,and the observation group received bladder muscle stimulation therapy in addition.The micturition diary and urodynamic indexes were assessed before and three months after treatment.Results After treatment,the frequency of urinary incontinence and the number of micturition in 24 hours,and residual urine volume significantly reduced(t>3.251,P<0.01),the urine output every time significantly increased (t>5.000, P<0.001), the maximum bladder capacity and urine flow rate increased (t>1.922, P<0.05),and the detrusor pressure in the end of filling period significantly reduced(t>8.302,P<0.001).All the indexes above were better in the observation group than in the control group(t>1.919,P<0.05). Conclusion Electrical muscular stimulation combined with the bladder func-tional training could improve the urination function in the patients with autonomic bladder after spinal cord injury.
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<p><b>OBJECTIVE</b>To investigate the effect of HBP-A on meniscal injuries and the expressions of genes associated with pathological hypertrophy and calcification of the meniscusinduced by abnormal loading.</p><p><b>METHODS</b>Bovine meniscus explants were subjected to 25% strain at 0.3 Hz for 3 h and treated with 0.6 mg/mL of HBP-A. The cell viability in the meniscus explants after 72 hin culture was determined using live/dead staining and the expression levels of genes associated with pathological hypertrophy and calcification of the meniscus (ANKH, ENPP1, ALP, MMP13, and IL-1) were measured using real-time PCR and Western blotting. The conditioned medium was collected for testing sulfated glycosaminoglycan (GAG) release.</p><p><b>RESULTS</b>The number of dead cells, loss of proteoglycan content, and the expressions of ANKH, ENPP1, ALP and MMP13, and IL-1 at both the mRNA and protein levels were all significantly lower in the meniscus explants treated with 0.6 mg/mL HBP-A than in the explants with only 25% abnormal pressure stimulation (n=3, P<0.05).</p><p><b>CONCLUSION</b>HBP-A can effectively alleviate meniscal injuries induced by abnormal loading and suppress the expressions of genes related with pathological hypertrophy and calcification of the meniscus, and can serve as a potential drug for treatment of knee osteoarthritis.</p>
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Animals , Cattle , Calcinosis , Drug Therapy , Glucans , Pharmacology , Hypertrophy , Menisci, Tibial , Osteoarthritis, Knee , Drug Therapy , Real-Time Polymerase Chain Reaction , Tibial Meniscus Injuries , Drug TherapyABSTRACT
<p><b>OBJECTIVE</b>To investigate the effects of serum containing Gumibao (Chinese character: see text) on the proliferation and differentiation of osteoblast induced by dexamethasone.</p><p><b>METHODS</b>Osteoblasts were extracted from skulls in newly born (within 24 hours) SD rats, and digested with collagenase. The first passage of cells were used for experiments. Cells were cultured in the medium containing different concentrations of dexamethasone (0, 10(-8), 10(-7), 10(-6), 10(-5) ,10(-4) mol/L). Alkaline phosphatase staining were carried out after 1 week and numbers of mineralized nodes with alizarin red staining were observed after 3 weeks. Accordingly, following the treatment of 10(-5) mol/L dexamethasone for 1 week, cells were cultured in the medium with serum containing Gumibao (Chinese character: see text). One week after Cumibao (Chinese character: see text) treatment, cells were stained with Alkaline phosphatase and collagen I and PCNA were examined by Western-blot. However, the observation of numbers of mineralized nodes with alizarin red stain required one more week.</p><p><b>RESULTS</b>High concentration of dexamethasone could inhibit the expression of PCNA, collagen I, alkaline phosphatase and reduce the number of mineralized nodes of osteoblast, while serum containing Gumibao (Chinese character: see text) could reverse the inhibition.</p><p><b>CONCLUSION</b>High concentration of dexamethasone could inhibit the proliferation and differentiation of osteoblastic cells, while serum containing Gumibao (Chinese character: see text) could reverse the inhibition.</p>
Subject(s)
Animals , Female , Rats , Cell Differentiation , Cell Proliferation , Cells, Cultured , Collagen Type I , Dexamethasone , Pharmacology , Dose-Response Relationship, Drug , Drugs, Chinese Herbal , Pharmacology , Osteoblasts , Cell Biology , Physiology , Proliferating Cell Nuclear Antigen , Rats, Sprague-DawleyABSTRACT
Objective To further understand the biomechanical relationship between activities of cervical spine and blood flow of vertebral artery (VA) by developing the VA finite element model and calculating the fluid-structure interaction. Methods Based on the normal model of cervical spine and the developed C0-T1 finite element model with bilateral VA, the flexion and extension, right and left lateral bending, right and left axial rotation movement of cervical spine at physiological velocity were simulated. The effects of cervical activities on stress of vertebral arterial wall were observed, and the biomechanical interaction between the vessel wall and fluid was calculated by fluid-structure interaction equation to obtain the hemodynamic parameters. Results The maximum stress was usually concentrated on the both sides of C2 transverse foramen, where the second arc of vertebral arterial wall protruded into the cranial direction during cervical activities. The maximum strain of the vessel wall was most obvious during the extension and lateral bending movement, with strain ratio of 23.04% and 35.5%, respectively. The maximum stress on the vessel was located in the position of contralateral transverse foramen during lateral bending movement, while the maximum strain on the vessel was located in the position of ipsilateral transverse foramen during rotation movement. In aspect of cervical spine range of motion (ROM), the minimum volume flow rate occurred within 30%-40% of the physiological ROM. The volume flow rate-time curve of bilateral VA was similar during flexion and extension movement, when the circulation of flow rate was completed for two times within 0.5 s. The peak and valley of ipsilateral blood flow in volume flow rate-time curve occurred earlier than that of contralateral blood flow during lateral bending movement, while the results of rotation movement were opposite. Conclusions The obtained stress features of bilateral VA vessel and the law of the volume flow rate-time curve validated the experimental results with those in the literature, which could reasonably explain the clinical phenomenon. The established model would provide an ideal platform for researches on vertebral artery-related diseases.
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<p><b>OBJECTIVE</b>To analyze the current situation on China Infectious Disease Automated-alert and Response System (CIDARS) in Guangdong province, China.</p><p><b>METHODS</b>Early-warning signals and response time were analyzed by using three approaches of CIDARS. Positive rates of early-warning signals and error early-warning rates prior and after the adjustment,were compared.</p><p><b>RESULTS</b>Totally, 114 585 early-warning signals appeared, with an average response time of 1.35 hours from April 21, 2008 to December 31, 2012. There were 12,394 early-warning signals in terms of fixed threshold method with a positive rate of 7.96%. 85 727 early-warning signals appeared under the mobile percentile method with a positive rate of 0.85%. There were 16,464 early-warning signals by using accumulation and control chart methods,with a positive rate of 1.82%. Results showed there was a positive correlation between the number of reported cases and the number of early-warning signals (r = 0.924, P < 0.01). The overall positive rate in Guangdong province increased from 1.48% to 2.14%, after the adjustment done by the Chinese Center for Disease Control and Prevention in 2010. The error early-warning rates regarding eight infectious diseases including hepatitis A, bacillary or amebic dysentery, both typhoid and paratyphoid fevers had reduced.</p><p><b>CONCLUSION</b>The early-warning signal response appeared timely in CIDARS with good operation. However, despite the improvement on the efficacy of CIDARS, some functions and parameters of the systems still need to be adjusted.</p>
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Humans , China , Communicable Disease Control , Methods , Disease Outbreaks , Population Surveillance , MethodsABSTRACT
<p><b>OBJECTIVE</b>To compared the activity and yield rate of osteoblast obtained by different collagenase digestion methods, to find a better way to extract osteoblast for the experimental researches of osteoporosis.</p><p><b>METHODS</b>Ten 24-hour-old SD rats were were euthanized. The cranium of rats were removed and cuted into blocks of 1 mm x 1 mm size. After digested by trypsin for 15 min, all the cranium were divided into two equal parts, and randomly divided into two groups which would be digested by type I collagenase and type II collagenase separately for two times. The rat cells of the two groups were cultured in thermostat incubator with 5% CO2 under the condition of 37 degrees C. The primary culture osteoblasts were counted by using a haemacytometer after digestion and 72 hours later. The second generation osteoblasts cultured 48 h were dyed by NBT/BCIP staining solution, and were detected by quantitative measurement with PNPP.</p><p><b>RESULTS</b>The cells had irregular shapes. The results of cell counting showed that the cell number of type I group was larger than type 11 group. Alkaline phosphatase dyeing were positive. Detecting of alkaline phosphatase using the method of PNPP showed that the absorbance value in type I group were higher than type II group (P<0.05).</p><p><b>CONCLUSION</b>Two types of collagenase are both suitable for the in vitro culture of rat osteoblasts. The activity and yield rate of osteoblasts in type I group are higher which could provide more stable seed cells for the treatment of osteoporosis.</p>
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Animals , Female , Male , Rats , Cell Count , Cell Culture Techniques , Methods , Collagenases , Metabolism , Osteoblasts , Cell Biology , Rats, Sprague-DawleyABSTRACT
<p><b>OBJECTIVE</b>To investigate the regulatory effect of Jingang Jiangu pill (see text, JGJG) on expression of integrin in ovariectomized rats.</p><p><b>METHODS</b>Fifty ovariectomized 10 months old female rats were randomly divided into 5 groups: Fushanmei group (FSM), Jingang Jiangu pill (see text) group (JGJG), Gusongbao granule group (GSB), Model group (OVX), Sham group. After ovariectomized,the rats were raised in the same environment for 13 weeks. The rats in JGJG group took 0.13 g JGJG pill orally each day for each rat; the rats in GSB group took 0.86 g GSB granule orally each day for each rat; the rats in FSM group took 0.28 mg FSM orally each day for each rat; and the rats in OVX and sham groups took sodium. The treatment duration of rats in above 5 groups was 13 weeks. Bone mineral density (BMD) and the expression of integrin beta1 and alphavbeta3 were detected in each group after the treatment. RESYKTS: The BMD and the expression of integrin beta1 in FSM group, JGJG group and GSB group improved obviously than that of OVX group. There were statistical difference between these groups (P<0.05). The expression of integrin alphavbeta3 of the three treating groups significantly depressed.</p><p><b>CONCLUSION</b>The JGJG pill improves BMD and express of integrin beta1, in ovariectomized rats and reduces express of integrin alphavbeta3 through the regulation of the coupling of osteoblasts and osteoclasts.</p>
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Animals , Female , Rats , Bone Density , Disease Models, Animal , Integrin alphaVbeta3 , Integrin beta1 , Medicine, Chinese Traditional , Osteoporosis , Drug Therapy , Metabolism , Ovariectomy , Rats, WistarABSTRACT
<p><b>OBJECTIVE</b>To investigate the effects of osthole on proliferation of neonatal rat osteoblast and the mechanism.</p><p><b>METHODS</b>Ten 24 hours old SD rats were executed by dislocating. The cranium of rats were removed and cut into blocks of 1 mm x 1 mm size. After digested by trypsin for 15 min, the cranium were digested by type I collagenase for one hour two times. The mixed cells were cultured in thermostat incubator with 5% CO2 under the condition of 37 degrees C. To identify the cells, ALP staining and alizarin red staining were performed after cultured 48 h and 28 d. The osteoblasts were randomly divided into five groups. Cells were treated with osthole at concentrations of 100, 50, 25, 12.5, 0 micromol/L. CCK-8 method was used to evaluate the proliferation after 24 h,48 h and 72 h. The expression of PCNA and beta-catenin protein were detected through the method of Western Blot after one week.</p><p><b>RESULTS</b>The cells had irregular shapes and showed typical features of osteoblast. The results of ALP staining and alizarin red staining were both positive. CCK-8 detection showed that the osthole with final concentration of 100 micromol/L inhibited the proliferation of osteoblast after 24 h, while the osthole with final concentrations of 50 micromol/L and 25 micromol/L displayed the inhibition effect after 48 h. The osthole of 12.5 micromol/L had no obvious influence on the proliferation of osteoblast. The result of Western Blot showed that osthole reduced the expression of PCNA and beta-catenin protein in a dose-dependent manner.</p><p><b>CONCLUSION</b>The osthole with final concentrations of 100, 50, 25 micromol/L inhibited the proliferation of osteoblast (P < 0.05). The osthole with final concentrations of 12.5 micromol/L had no obvious influence on the proliferation of osteoblast (P > 0.05). These findings demonstrate that osthole may inhibit the proliferation of osteoblast by regulating the Wnt/beta-catenin signaling in osteoblast.</p>
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Animals , Female , Male , Rats , Animals, Newborn , Cell Differentiation , Cell Division , Cell Proliferation , Cells, Cultured , Coumarins , Pharmacology , Osteoblasts , Cell Biology , Metabolism , Rats, Sprague-Dawley , Signal Transduction , Sincalide , Metabolism , beta Catenin , MetabolismABSTRACT
Objective To understand the infection status of pandemic 2009 H1N1 influenza after the first epidemic wave and to estimate the infected population. Methods Multi-stage stratified random sampling was introduced with 4500 subjects chosen in Guangdong province. 1500 people were selected from 5 districts (3 streets were selected in every district) in Guangzhou city which was representing the large cities. 1500 people were respectively selected from medium-sized city and rural areas, including 20 cities (1 county or district was selected in every city, at least 1 street or town was selected in every county or district respectively and then 1-2 residential area or county was selected in every street or township, respectively). Every sample was selected in accordance with the principle of random sampling, excluding those who had injected with novel H1N1 vaccine. We used hemagglutination inhibition test to understand the serum antibody level of novel H1N1, with title of 1∶40 as positive. Results A number of 4319 specimens, distributed in 21 cities, 25 counties, 85 streets or townships, 144 residential areas, were tested, with an overall positive rate as 22.82% (985/4319). The positive rate of those who had no symptoms of cold since June was 23.47% (471/2007).The positive rate of those who had fever, cough or sore throat was 26.25% (714/2720). The positive rate of those who had influenza-like illness (ILI) was 29.69% (337/1135 ). Conclusion The infection rate in the first epidemic wave of the novel influenza A (H1N1) pandemic in Guangdong province was 22.82%(985/4317). Based on the number of residonts in Guangdong province, the number of natural was estimated to have reached 21.78 million.
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<p><b>OBJECTIVE</b>To analysis the risk factors influencing nosocomial infection of severe acute respiratory syndrome (SARS) in health-care workers and to evaluate effectiveness of its control and preventive measures in 13 key hospitals caring for SARS patients.</p><p><b>METHODS</b>Number of SARS patients, clinical conditions of them, its attack rate in health-care workers, and characteristics of hospitals, including their environment, isolating measures, etc. were investigated at the 13 hospital in Guangzhou to analyze the risk factors influencing nosocomial infection of SARS and its attack rates in health-care workers before and after implementation of preventive measures and to evaluate their effectiveness.</p><p><b>RESULTS</b>Totally, 841 patients with SARS were treated at the 13 hospitals in Guangzhou and 285 health-care workers caring for them infected nosocomially. Attack rate in health-care workers was higher at general hospitals, hospital accepting more cases in critical conditions and hospitals with poor precautious measures, and lower in hospitals with isolated wards or areas, or department of infection, specially caring for SARS patients, and those with effective intervention measures to prevent secondary infection.</p><p><b>CONCLUSION</b>Nosocomial infection of SARS in health-care workers was affected by clinical condition of SARS patients, characteristics and environment of hospitals and their personal protective measures adopted.</p>
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Humans , China , Epidemiology , Contact Tracing , Cross Infection , Hospitals , Occupational Exposure , Personnel, Hospital , Severe Acute Respiratory Syndrome , EpidemiologyABSTRACT
<p><b>OBJECTIVE</b>To evaluate the effectiveness of personal protective measures of health care workers (HCWs) against severe acute respiratory syndrome (SARS).</p><p><b>METHODS</b>A case-control study from ten hospitals in Guangdong, with 180 non-infected and 77 infected staff members that accessed the isolation unit every day, and participated in direct first aid for severe SARS patients. All participants were surveyed about how they were using personal protective equipment (PPE), protective drugs and hygiene habits when caring for patients with SARS. Statistical analysis was done with either chi(2) or Fisher's exact test for univariate analysis, whereas we used forward stepwise selection (Waldesian) for logistic regression.</p><p><b>RESULTS</b>Univariate analysis showed that mask, gown, gloves, goggles, footwear, "hand-washing and disinfecting", gargle, "membrane protection", "taking shower and changing clothing after work", "avoid from eating and drinking in ward", oseltamivir phospha tall had protective effects (P < 0.05), but stepwise logistic regression showed significant differences for mask (OR = 0.78, 95% CI: 0.60 - 0.99), goggles (OR = 0.20, 95% CI: 0.10 - 0.41) and footwear (OR = 0.58, 95% CI: 0.39 - 0.86). Analysis for linear trend in proportions showed that dose response relationship existed in mask, gown, gloves, goggles, footwear, gargle, "membrane protection" and "taking shower and changing dree after work" (P < 0.01). The attack rate of HCWs who were rescuing severe SARS patients without any PPE was 61.5% (16/26). It seemed that the more the protective measures were used, the higher the protective effect was (P < 0.001), and could reach 100% if mask, gown, gloves, goggles, footwear, "hand-washing and disinfecting" were all used at the same time.</p><p><b>CONCLUSIONS</b>Nosocomial infection of SARS can be prevented effectively by precautions against droplets and personal contact. HCWs must take strict protection according to the guidance of WHO or Chinese MOH and pay attention to personal hygiene.</p>
Subject(s)
Female , Humans , Male , China , Cross Infection , Health Occupations , Education , Logistic Models , Protective Devices , Classification , Severe Acute Respiratory Syndrome , Surveys and QuestionnairesABSTRACT
<p><b>OBJECTIVE</b>To explore epidemiological features and risk factors of severe acute respiratory syndrome (SARS) in Guangdong Province of China, so as to work out effective strategies for its better control.</p><p><b>METHODS</b>A total of 1 511 clinically confirmed SARS cases in Guangdong Province of China from November 16, 2002 to Jun 15, 2003 were retrospectively analyzed.</p><p><b>RESULTS</b>The first SARS case was identified in Foshan municipality on November 16, 2002, followed by 1 511 clinically confirmed cases (including 58 deaths) up to May 15, 2003. Of all cases, health care workers and community family cluster cases accounted for 19.38% and 12.04%. 65.86% SARS patients aged 20 - 49 years, and increased incidence was positively related to their ages. 95.97% cases lived in the following five cities around Pearl Delta Area: Foshan, Guangzhou, Shenzhen, Zhongshan, and Jiangmen. Eleven early reported cases in the communities took animal-related positions. Face-to-face contacts with infected droplets were the main transmission route. An epidemic peak occurred during January 28 to February 26, and those cases accounted for 50.69% of total. Incidence, mortality, and case fatality of SARS were 1.77/100,000, 0.07/100,000, and 3.84% respectively. The mean incubation period was 4.5 days.</p><p><b>CONCLUSION</b>The most effective way to control SARS is to break the chain of transmission from infected to healthy persons-early identification, prompt and effective isolation, and vigorous close contact tracing. Hospital infections among health care workers is critical. Several observations support the hypothesis of an animal origin for the disease.</p>