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ObjectiveBy analyzing the current situation of drug selection and evaluation in medical institutions in the world，we aim to understand the development of relevant selection methods and tools，provide reference basis for drug selection in traditional Chinese medicine （TCM） medical institutions，and promote the optimization of drug catalogs in TCM medical institutions. MethodBased on the method of scoping review，the eight databases were systematically searched，the included documents were screened，extracted and analyzed，and the research results were graphically displayed. ResultA total of 23 articles were included in this study，including 13 in Chinese and 10 in English，involving 23 methods or tools related to drug selection. Of the 14 methods or tools from Chinese medical institutions，the earliest one was published in 2012，and five were published in 2022. The published methods or tools involved different levels of hospitals，different drug varieties，different evaluation angles，etc.，such as the drug selection methods of one county hospital and one township hospital， methods and tools for different types of drugs such as antibacterial drugs，ibuprofen preparations，proton pump inhibitors and hypoglycemic drugs used in pediatric intensive care units， Chinese patent medicine selection tools， tools for evaluation from the perspective of pharmacoeconomics， and universal tools for selecting domestic medical institutions. The nine drug selection tools of foreign medical institutions were from the European，American，Asian and African countries. It was first published in 1955. The contents included the formulation standards that medical institutions should follow for drug prescription sets，the management formulation and update of hospital prescription sets，and drug evaluation tools. ConclusionOn the whole，the drug selection methods and institutional methods of foreign medical institutions developed earlier than those in China. In recent years，Chinese medical institutions have paid high attention to drug selection and released various types of drug selection tools. However，the standardization should be further improved in the future.
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<p><b>OBJECTIVE</b>To evaluate the effect of demineralized cancellous bone (DCB) seeded with allogeneic chondrocytes for repairing articular osteochondral defects in rabbits.</p><p><b>METHODS</b>Articular chondrocytes were isolated from a 1-month-old male New Zealand rabbit for primary culture. The passage 1 chondrocytes were seeded onto prepared rabbit DCB scaffold to construct tissue-engineered cartilage and cultured for 2 weeks. Full-thickness articular osteochondral defects (3 mm both in diameter and depth) were created on both sides of the femoral medial condyles in 30 New Zealand white rabbits (age 4- 5 months). In 20 of the rabbits, the defects were filled with the tissue-engineered cartilage on the right side (group A) and with DCB only on the left side (group B); the remaining 10 rabbits did not receive any implantation in the defects to serve as the control (group C). At 1, 3, and 6 months after the implantation, tissue samples were collected from the defects for macroscopic observation and histological examination with Toluidine blue (TB) and collagen type Ⅱ staining. The effect of defect repair using the tissue-engineered cartilage was assessed at 6 months based on the histological scores.</p><p><b>RESULTS</b>The prepared DCB had a spongy 3D structure with open and interconnected micropores of various sizes and showed good plasticity and mechanical strength. DCB began to degrade within 1 month after implantation and was totally absorbed at 3 months. At 6 months after implantation, the defects filled with the chondrocyte-seeded DCB were repaired mainly by hyaline-like cartilage tissues, which were well integrated to the adjacent cartilage without clear boundaries and difficult to recognize. The chondrocytes were located in the lacunate and arranged in vertical columns in the deep repaired tissue, where matrix proteoglycans and collagen type Ⅱ were distributed homogeneously close to the normal cartilage. The subchondral bone plate was reconstructed completely. The defects implanted with DCB only were filled with fibrocartilage tissue, as compared with fibrous tissue in the control defects. The histological scores in group A were significantly superior to those in group B and C ( < 0.05), but the scores for subchondral bone plate reconstruction were comparable between groups A and B at 6 months.</p><p><b>CONCLUSIONS</b>DCB is a good scaffold material for preparing tissue-engineered cartilage, and chondrocyte- seeded DCB can repair articular osteochondral defects by inducing the generation of hayline-like cartilage.</p>
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Clinical pharmacist participated in the whole process of drug therapy for one case of acute hypotension adverse event af-ter PCI. Based on the patient's preoperative and postoperative medication, the hypothesis was proposed and the causes of acute hypo-tension were positively analyzed. From the perspective of pharmacology, professional advice was given and satisfactory therapeutic effects were obtained. The experience of pharmaceutical care after PCI was summarized as well.
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Objective To clarify the role of nuclear factor κB(NF-κB) signaling pathway in pathogenesis of Kashin-Beck disease(KBD) by observing the expression of NF-κB p65 in the whole blood samples of patients with KBD and controls,and the expression of NF-κB p65 in C28/I2 chondrocyte, and to analyze the role of NF-κB p65 molecule in chondrocyte apoptosis. Methods Through a case-control study, 161 patients with KBD (KBD group) were selected from Xunyi, Yongshou, Changwu, Linyou, Qianyang and Long counties in KBD endemic areas and 312 healthy people(control group) were matched by age and sex in Shaanxi Province. Venous blood samples were collected from patients and healthy controls, which were anticoagulated and used for determination of NF-κB p65 protein.According to the group design,the model of C28/I2 chondrocyte oxidative damage was established.The experiments were divided into 4 groups including control group(C), tBHP injury group (O, tBHP 300.00 μmol/L), low selenium pre-protection group (OS1, 0.05 mg/L Na2SeO3+ 300.00 μmol/L tBHP), and middle selenium pre-protection group(OS2, 0.10 mg/L Na2SeO3+ 300.00 μmol/L tBHP). Then, cell apoptosis was detected by Hoechst 33342 and reactive oxygen species (ROS) was detected by dichlorfluorescein(DCF) method. The protein was extracted by Trizol method, then protein expression level of NF-κB p65 molecule was detected by Western blotting in whole blood samples and C28/I2 chondrocyte. Results The differences in age and sex were not statistically significant between KBD group and control group (t = 0.336, P > 0.05; χ2= 0.407, P > 0.05). The protein expression level of NF-κB p65 in KBD group was 1.835 times as high as that of control group (KBD:0.167 ± 0.026, control: 0.091 ± 0.014, t = 5.147, P < 0.01). Under the fluorescence microscope, chondrocyte showed strong blue fluorescence in tBHP group and the level of ROS(1.219 ± 0.104) was higher than those of low and middle selenium pre-protection groups(0.832 ± 0.077, 0.635 ± 0.070, P < 0.05).The protein expression level of NF-κB p65 in tBHP group (1.563 ± 0.351) was higher than that of control group (0.451 ± 0.069, P < 0.05), and protein levels of NF-κB p65 had a decreasing tendency in low and middle selenium pre-protection groups compared to tBHP group. Conclusion The NF-κB signaling pathway is up-regulated in KBD patients, moreover, chondrocyte experiments show that cell apoptosis is mediated via upregulation of NF-κB p65,which suggests NF-κB signaling pathway may play an important role in pathogenesis of KBD.
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Objective To explore the effects of oxidative damage and selenium on the apoptosis of articular chondrocytes and the expression of selenoprotein genes.Methods C28/I2 chondrocytes were preincubated for 24 h,using sodium selenite (Na2SeO3) or t-butyl hydroperoxide (tBHP) for 24 h.The experiment was divided into six groups,including control group (C,0.00 mg/L Na2,SeO3 + 0.00 μmol/L tBHP),selenium beforehand protection group (S2,0.10 mg/L Na2SeO3),oxidative damage group (O,150.00 μmol/L tBHP),low dose selenium protection group (OS 1,0.05mg/L Na2SeO3 + 150.00 μmol/L tBHP),medium dose selenium protection group (OS2,0.10 mg/L Na2SeO3 + 150.00 μmol/L tBHP),and high dose selenium protection group (OS3,0.15 mg/L Na2SeO3 + 150.00 μmol/L tBHP).After 24 h,Hoechst 33342 staining method was used to observe apoptosis,mRNA expression of glutathione peroxidase 1 (GPX1),GPX4,deiodinase 2 (DIO2),DIO3,selenoprotein P (SEPP1),thioredoxin reductase 1 (TrxR-1) and selenoprotein W(Sel W) was detected by Real-time PCR,both experiments were done three times.Results Apoptotic rates of C,S2,O,OS1,OS2,OS3 groups [(0.78 ± 0.06)％,(13.61 ± 7.11)％,(92.27 ± 3.44)％,(71.38 ± 5.22)％,(44.31 ± 9.16)％,(72.46 ± 4.69)％] were compared between groups,the differences were statistically significant (F =120.10,P ＜ 0.01).The apoptotic rates of O group was significantly higher than that of C group (P ＜ 0.05);compared to O group,the apoptotic rates of OS1,OS2,OS3 groups decreased significantly (P＜ 0.05),OS2 group was the most obvious.DIO2,SEPP1,GPX1,GPX4,TrxR-1,Sel W mRNA levels were compared in the six groups,the differences were statistically significant (F =24.60,14.53,127.60,30.60,637.10,59.64,P ＜ 0.01).Compared to C group (1.00 ± 0.00),the mRNA levels of GPX1 (0.10 ± 0.05),GPX4 (0.43 ± 0.09),TrxR-1 (0.11 ± 0.05) and Sel W (0.72 ± 0.15) in O groups were decreased significantly (P ＜ 0.05);compared to 0 group,the mRNA levels of GPX1 in OS1 (0.20 ± 0.03),OS2 (0.74 ± 0.10),and OS3 (0.30 ± 0.07) were increased significantly (P ＜ 0.05).Conclusion Down-regulated expression of selenoprotein genes are involved in the regulation process of articular cartilage apoptosis caused by oxidative stress,selenium also has a regulatory role in selenoprotein gene expression in articular chondrocytes.
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Objective To explore the effects of oxidative damage and selenium on the apoptosis of articular chondrocytes and the expression of selenoprotein genes.Methods C28/I2 chondrocytes were preincubated for 24 h,using sodium selenite (Na2SeO3) or t-butyl hydroperoxide (tBHP) for 24 h.The experiment was divided into six groups,including control group (C,0.00 mg/L Na2,SeO3 + 0.00 μmol/L tBHP),selenium beforehand protection group (S2,0.10 mg/L Na2SeO3),oxidative damage group (O,150.00 μmol/L tBHP),low dose selenium protection group (OS 1,0.05mg/L Na2SeO3 + 150.00 μmol/L tBHP),medium dose selenium protection group (OS2,0.10 mg/L Na2SeO3 + 150.00 μmol/L tBHP),and high dose selenium protection group (OS3,0.15 mg/L Na2SeO3 + 150.00 μmol/L tBHP).After 24 h,Hoechst 33342 staining method was used to observe apoptosis,mRNA expression of glutathione peroxidase 1 (GPX1),GPX4,deiodinase 2 (DIO2),DIO3,selenoprotein P (SEPP1),thioredoxin reductase 1 (TrxR-1) and selenoprotein W(Sel W) was detected by Real-time PCR,both experiments were done three times.Results Apoptotic rates of C,S2,O,OS1,OS2,OS3 groups [(0.78 ± 0.06)％,(13.61 ± 7.11)％,(92.27 ± 3.44)％,(71.38 ± 5.22)％,(44.31 ± 9.16)％,(72.46 ± 4.69)％] were compared between groups,the differences were statistically significant (F =120.10,P ＜ 0.01).The apoptotic rates of O group was significantly higher than that of C group (P ＜ 0.05);compared to O group,the apoptotic rates of OS1,OS2,OS3 groups decreased significantly (P＜ 0.05),OS2 group was the most obvious.DIO2,SEPP1,GPX1,GPX4,TrxR-1,Sel W mRNA levels were compared in the six groups,the differences were statistically significant (F =24.60,14.53,127.60,30.60,637.10,59.64,P ＜ 0.01).Compared to C group (1.00 ± 0.00),the mRNA levels of GPX1 (0.10 ± 0.05),GPX4 (0.43 ± 0.09),TrxR-1 (0.11 ± 0.05) and Sel W (0.72 ± 0.15) in O groups were decreased significantly (P ＜ 0.05);compared to 0 group,the mRNA levels of GPX1 in OS1 (0.20 ± 0.03),OS2 (0.74 ± 0.10),and OS3 (0.30 ± 0.07) were increased significantly (P ＜ 0.05).Conclusion Down-regulated expression of selenoprotein genes are involved in the regulation process of articular cartilage apoptosis caused by oxidative stress,selenium also has a regulatory role in selenoprotein gene expression in articular chondrocytes.
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OBJECTIVE:To provide reference for the classification of the specification grade of Dioscorea opposite.METHODS:Using the Delphi method,17 experts,who were associated with the study of the distribution and clinical application of D.opposite,were selected to conduct two rounds of consultation on 18 samples of medicinal herbs.The importance of the sensory evaluation indexes of D.opposite was screened and the overall satisfaction was evaluated.The specification grade of D.oppositewas determined preliminarily.RESULTS:Totally 17 questionnaires were issued for each round of consultation,and 17 were recovered with recovery rate of 100％.According to statistical analysis,the average value of expert's authority coefficient was 0.77 +0.07,and average recurrence rate of individual review was (91.18 + 7.64)％.The average recurrence rates of group review in two rounds of consultation were (66.67 + 13.50)％ and (65.97 + 14.01)％.The influence of working life,business background and educational background on recurrence rates of group review for interviewed experts was comprehensive.The full score ratio of appearance and cross-sectional characteristics importance was more than 80％,and the average value was more than 0.8.The full score ratio and average value of iron yam were higher.CONCLUSIONS:It is accurate and scientific to evaluate the specification grade of Chinese medicinal herbs with sensory experience.The most important evaluation indicators of D.opposite were the appearance,shape and cross-sectional characteristics.And it is divided into iron yam and non-iron yam preliminarily.The iron yam grade in descending order:Henan sandy iron yam,Shandong sandy iron yam,Henan loquat iron yam.The non-iron yam grade in descending order:Henan Huaiqingfu D.opposite,Hebei Xiaobaizui D.opposite,Hebei Ma D.opposite and Shanxi Chang D.opposite.
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Objective:To discuss the importance and significance of training base establishment for clinical pharmacy of traditional Chinese medicine ( TCM) . Methods:The urgency and necessity of training base establishment for TCM clinical pharmacy was stated based on the status and problems of TCM clinical pharmacy, and the important significance of training base establishment of TCM clini-cal pharmacy was clearly put forward. Results:The establishment of TCM clinical pharmacy training base with the aim of training pro-fessional talents for TCM clinical pharmacy can provide clinical pharmaceutical care for patients with higher quality, which surely will promote the development of TCM clinical pharmacy and the pharmaceutical care level. Conclusion:It is extremely urgent and signifi-cant to set up TCM clinical pharmacy training base, which is the important topic needed to be solved in TCM clinical pharmacy work.
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In order to ef fectively control the quality ofSophora lfos carbonisatus (flower and flower buds), this study established quality control methods and standard of the decoction pieces. Referring to the related methods in appendix of Chinese Pharmacopoeia (2010 Edition), the moisture, total ash, acid insoluble ash, alcohol extracts ofSophora lfos carbonisatus were measured, respectively, with rutin, quercetin as control substance. The eluents for rutin and quercetin are ethyl acetate - formic acid - water (8: 1:1) and chloroform - methanol - water (6.5:1:1), respectively and all TLC plates were observed at 365 nm. Total flavonoids are measured by visible - UV - spectrophotometric, and rutin and quercetin were determined by HPLC. The chromatographic conditions for rutin are: Kromasil C18 as the stationary phase, methanol -1% acetic acid (32:68) as mobile phase, flow rate: 0.8 mL·min-1, detection wavelength 257 nm,the column temperature 35℃; for quercetin: Kromasil C18 as the stationary phase, methanol -0.4% acetic acid (44:56) as the mobile phase, flow rate: 0.8 mL·min-1, detection wavelength 257 nm, the column temperature 40℃.The contents of moisture, total ash, acid insoluble ash, should not exceed 6%, 16%, 8.0% in flower and not exceed 6.0%,9.0%, 1.5% in buds, respectively. Under the conditions of TLC, in flower and flower buds, 2 reference substances can be separated well with others. Extract, total flavonoids, rutin, quercetin were no lower than 40.0%, 5.0%, 2.5%,0. 2% in flower and no lower than 45.0%, 10.0%, 5.0%, 0.9% in buds, respectively. The established standards can improve the levels of quality control, provide experimental data for safety and efficacy of clinical application of Sophora flos carbonisatus, and also offer supporting data for the Chinese Pharmacopoeia 2015 Edition.
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Objective To investigate the expressions of interleukin-1β(IL-1β), interleukin-6(IL-6) and tumor necrosis factor alpha(TNF-α) in cartilage of children with Kashin-Beck disease(KBD) in order to provide a possible mechanism of the disease. Methods Articular cartilage tissues of 5 KBD children(KBD group) were selected from KBD children autopsy samples keeping in Institute of Endemic Diseases, Medical School of Xi’an Jiaotong University; articular cartilage tissues of 5 normal children ( control group ) were selected from non-KBD areas of Shaanxi Province, three cases were from accident death children, two cases were the samples of congential malformation of six finger. Expressions of IL-1β, IL-6 and TNF-α in the cartilage were detected using immunohistochemistry; the cells of articular cartilage were divided into three areas (superficial zone, middle zone and deep zone) to analyze the expressions of IL-1β, IL-6 and TNF-α. Results The expressions of IL-1β in superficial zone , middle zone and deep zone of articular cartilage of KBD group (63.50 ± 7.19, 54.75 ± 5.50, 66.20 ± 9.91) were significantly higher than those of control group(5.75 ± 1.26, 0.00 ± 0.00, 0.00 ± 0.00, all P<0.05). The expression of IL-6 in superficial zone of articular cartilage in KBD group(55.25 ± 6.24) was significantly higher than that of control group(0.00 ± 0.00, P<0.05). The expressions of TNF-αin all zone of articular cartilage of KBD group(33.25 ± 6.50, 3.75 ± 0.96, 29.80 ± 1.92) were significantly higher than those of control group (3.74 ± 0.82, 0.00 ± 0.00, 0.00 ± 0.00, all P < 0.05). Conclusion The levels of IL-1β, IL-6 and TNF-α are up-regulated in articular cartilage of KBD children, suggesting that cytokines may play an important role in matrix degradation in KBD children cartilage.
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Objective To investigate the expressions of matrix metalloproteinases(MMPs) in Kashin-Beck disease(KBD) cartilage as well as in a KBD rat model of T-2 toxin poisoning under selenium deficient conditions, and to investigate the effect of T-2 toxin on MMP-13 expression in human chondrocytes in vitro in order to determine a possible mechanism underlying KBD. Methods Samples of articular cartilage were divided into 2 groups:controls(samples from 5 normal children, traffic accident or operation), and KBD(samples from 5 children with KBD, auctopsy). Thirty-two Sprague-Dawley rats were divided into two groups by body weight using random number table: normal diet group(n = 16) and selenium-deficient diet group(n=16). The selenium level in normal diet was 101.500μg/kg, and in selenium-deficient diet was 1.118μg/kg. Rats were fed for 4 weeks with selenium-deficient or normal diet, respectively. After successful build up of the low selenium rat model, normal diet group was then subdivided into 2 sub-groups: normal group(n = 8) and normal diet plus low T-2 toxin group(n = 8);and selenium-deficient diet group was also subdivided into 2 sub-groups: selenium-deficient group ( n = 8 ) and selenium-deficient diet plus T-2 toxin group ( n = 8 ) . T-2 toxin of 100 μg·kg-1·d-1 was administered by intragastric administration for 30 days. Then the rats were sacrificed, and their knee joints were processed for histopathological evaluation. MMP-1 and MMP-13 locations in cartilages were performed by inmmunohistochemistry. Human chondrocytes C28/I2 were cultured in vitro. The experiment was divided into 4 groups: empty vector plasmid group, MMP-13 promoter plasmid group, MMP-13 promoter plasmid plus 20 μg/L T-2 toxin group and MMP-13 promoter plasmid plus 40 μg/L T-2 toxin group. MMP-13-luciferase reporter plasmid and vector plasmid were transiently transfected into C28/I2 cells for 24 hours, and then treated with 20 - 40 μg/L T-2 toxin for 24 hours. Transactivation of human MMP-13 promoter was analyzed using luciferase reporter constructs containing sequences spanning-1602 to+20 bp in C28/I2 chondrocytes. Results The percentages of chondrocytes staining for MMP-1 in the superficial and middle zones of KBD samples [(29.73 ± 10.12)%, (28.27 ± 0.91)%] were significantly higher than those of controls[(2.47 ± 0.11)%, (0.00 ± 0.00)%, all P < 0.05]. The percentages of chondrocytes staining for MMP-13 in the superficial and middle zones of KBD samples [(13.21 ± 4.32)%, (41.85 ± 6.32)%] were significantly higher than those of controls[(5.72 ± 0.31)%, (0.00 ± 0.00)%, all P<0.05]. The percentages of chondrocytes staining for MMP-13 in the superficial and middle zones of rats fed with selenium-deficient diet plus T-2 toxin group[(13.21 ± 4.32)%, (61.85 ± 8.68)%] were significantly higher than those of the normal and selenium-deficient groups[(2.43 ± 0.22)%, (5.89 ± 0.69)%, (3.03 ± 0.29)%, (25.99 ± 0.57)%, all P < 0.05]. Moreover, T-2 toxin activated the MMP-13 promoter detected with luciferase reporter assays in C28/I2 cells. The luciferase activities in MMP-13 promoter plasmid plus 20 μg/L T-2 toxin group and MMP-13 promoter plasmid plus 40μg/L T-2 toxin group(0.082 78 ± 0.008 40, 0.103 35 ± 0.013 19) were significantly higher than those in empty vector plasmid group and MMP-13 promoter plasmid group(0.024 19 ± 0.000 96, 0.040 32 ± 0.003 56, all P < 0.05). Conclusions These data suggest that T-2 toxin induces cartilage matrix degradation through up-regulation of MMP-13 promoter expression. Increased MMPs staining intensity in KBD cartilage and the rat KBD model of T-2 toxin poisoning under selenium deficient conditions suggest that matrix degradation appear to be driven by MMPs activity.
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<p><b>OBJECTIVE</b>To study the chemical constituents from Centipeda minima.</p><p><b>METHOD</b>The compounds were separated and purified by silica gel, reserved phase silica gel and Sephadex LH-20 gel column chromatography. Their structures were identified on the basis physicochemical property and spectral analysis.</p><p><b>RESULT</b>Four chemical constituents were separated from ethyl acetate extracts and five chemical constituents were separated from n-butanol extracts of C. minima and identified as (2R,3R)-(+)-7,4'-di-O-methyldihydrokaempferol (1), iristectorin A (2), 4',5,8-trihydroxy-7-methoxyisoflavone (3), 3-trimethoxyquercetin (4), 3-O-caffeoyl-alpha-glueopyranose (5), 3-O-caffeoyl-beta-glucopyranose (6), quercetin (7), epipinoresinol (8) and hispidulin (9).</p><p><b>CONCLUSION</b>Compounds 1-3, 5-6, 8 and 9 are separated from this genus for the first time.</p>
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Asteraceae , Chemistry , Drugs, Chinese Herbal , Chemistry , Molecular Structure , Quercetin , ChemistryABSTRACT
Objective To examine matrix proteoglycan metabolic markers and probe into the turnover of matrix proteoglycan and enzyme-mediated role of matrix metalloproteinases (MMPs) and aggrecanases in reparative tissues with tissue engineering cartilage. Methods Tissue-engineered cartilage was constructed by cancellous bone matrix gelatin (BMG) with allogeneic chondrocytes in vitro for 2 weeks, then implanted to repair osteochondral defects of rabbit knee joint. Samples were obtained 6 months later to explore the expressions of 3-B-3(-) epitope, MMPs, MMP-generated epitope BC-4 and aggrecanases-generated epitope BC-13. Results In repaired tissues, the expression of 3-B-3(-) epitope increased, but that of MMPs and MMP-generated epitope BC-4 reduced. There was no expression of aggrecanases-generated epitope BC-13. Conclusion Expressions of 3-B-3(-), MMPs, BC-4 and BC-13 can help probe into the matrix proteoglycan turnover in reparative cartilage tissues. Anabolism exceeds catabolism in the repaired tissues. MMPs play an important role in the conservative baseline turnover of proteoglycan and remodeling of the graft tissues.
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OBJECTIVE To explore a target mornitoring method for hospital infection control based on web. METHODS According to hospital information system(HIS),using the database oracle 10.0 and progamming tools powerbiuld 9.0,based on inserted catheter and project,the objective mornitoring sofeware of hospital infection were explored and developed. The real-time mornitoring system was established for the important departments and projects of the Hospital infection. RESULTS By using HIS for sharing data and extracting useful data from the database and the main line of inquiry,the results of automatic statistics.The print function was convenient for extracting the basic materials of the subjiects under monitoring and helpful to supervise actively. The export capabilities could facilitate further analysis of data processing. CONCLUSIONS The mornitoring of the HIS flow is improved,A bridge were set up between infectious mornitoring and the clinic department. We can get a complete basic information,a comprehensive monitoring object and realize the standardization of information,aquire data easy and accurate,determine the dynamic observation in a timely manner. the humanpower and material resources have been saved.
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BACKGROUND: Deficit of nivalenol (NIV) and selenium (Se) is related with kashin-beck disease (KBD) to certain extent. Hyaluronic acid (HA) metabolism affects directly the polymerization of proteoglycans (PG) and normal structure and function of cartilage. The integration with HA receptor on surface of cartilage tissue is the key link in HA metabolism. Being the main receptor of HA on chondrocytic membrane, CD44 expression impacts directly HA metabolism, further affects cartilage matrix metabolism, which is extremely important to maintaining the structure and function of cartilage matrix.OBJECTIVE: To probe into the injury and protection of related etiology of KBD to target tissue cells and the mechanism on degenerative necrosis of chondrocytes.DESIGN: Randomized controlled observation was designed.SETTING: Department of Genetics and Molecular-Biology of Medical College of Xi' an Jiaotong University.MATERIALS: The experiment was performed in Key Laboratory Room on Ministry of Education associated with Environment and Disease of Xian Jiaotong University from October 2002 to July 2004. One New Zealand pedigree young rabbit aged 30 days was employed and its humerus, femurs and tibia were cut out in surgery.METHODS: With cell culture, the model of bone tissue was reconstructed in vitro, in which, NIV of various concentrations, KBD suspicious infectious agent and Se, the protective factor were added. HA receptor CD44 on chondrocytic membrane and soluble CD44 in cell culture solution were determined.MAIN OUTCOME MEASURES: ① Microscopic observation of adhesion molecule CD44 on chondrocytic surface. ② Soluble CD44 in chongrocytic culture solution.RESULTS: ① Microscopic observation of adhesion molecule CD44 on chondrocytic surface: CD expression in chondrocytic membrane was decreased with increasing of NIV concentration and it was in tendency of increasing with Se added. ② Soluble CD44 in chongrocytic culture solution:The concentration of soluble CD44 in cell culture solution was decreased gradually following the increased concentration of NIV, but it was increased in high concentration group and such tendency did not alter when Se added. Except blank control and Se control, significant difference was presented among groups (P ＜ 0.05).CONCLUSION: NIV disturbs adhesion molecule CD44 expression on chondrocytic surface and further induces metabolic disturbance of cartilage extracellular matrix. Se supplementation can resist the injury of NIV to chondrocytes, but its action is limited.
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In this paper, We present the method of using glucuronate as a parameter to determine the growth of cartilage cell in the culture media. Using this method, we observed the metabolic condition of cartilage cell and knew the effect of the serum with the various types and different concentration to the growth of cartilage cell. The. experimental method was used for the first passage chondrocyte of rabbit. We put the cartilage cell in the culture media containing different concentration of serum from calf and human; then, observed the growth of cartilage cell under the microscope, collected the medium in given time; and determined the amount of glucuronate in the medium. Our results indicated that the first passage chondrocyte of rabbit can grow in the medium containing serum of calf and human in 5%～15% Also, the study shows that it is possible to determine the amount of glucuronate in the medium as a parameter to respond the growth of the cartilage cells.
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The experiment was carried out on rats fed with synthetic diet containing torula yeasts with different levels of selenium, we collected their urine of 24 hours before the test on the 30th day and the 40th day of the test respectively. The level of acid glycosaminolycan in urine was determined by precipitate carbazole sulfate with zirconium oxychloride (ZrOCl_2) to measure glucuronate. The results by F test showed no significant difference in each group. The following two points have been reached. (a) The glucuronate level general rises in each group on the 30th day and declines on 40th day, especially in the low selenium group.(b) The more selenium the diet contains, the more glucuronate is excreted. The level of glucuronate in urine is correlated with that of blood selenium, with no significant difference.