Current status and prospect of biomarker research for schizophrenia / 中华检验医学杂志

Schizophrenia is a serious mental disease. The diagnosis of schizophrenia so far relies heavily on subjective evidence, including self-reported experiences by patients, manifestations described by relatives, and abnormal behaviors assessed by psychiatrists. The diagnosis, monitoring of the disease progression and therapy efficacy assessment are challenging due to the lack of established laboratory biomarkers. Based on the current literature, clinical consensus, guidelines, and expert recommendations, this review highlighted evidence-based potential laboratory biomarkers for the diagnosis of schizophrenia, including genetic biomarkers, neurotransmitters, neurodevelopmental-related proteins, and intestinal flora, and discussed the potential future directions for the application of these biomarkers in this field, aiming to provide an objective basis for the use of these biomarkers in the early and accurate diagnosis, treatment, and prognosis and rehabilitation assessment of schizophrenia.

Current status and correlates of screen time among children in Guangdong during the COVID-19 pandemic / 中国学校卫生

@#To investigate the current status of online classes, screen time and its influencing factors among primary school students in Guangdong during the 2019 novel coronavirus pandemic.@*Methods@#Using the convenience sampling method, a total of 5 266 pupils aged 6-12-years-old from Guangzhou, Zhanjiang, and Zhongshan participated in the online questionnaire. ANOVA or chi square tests were performed to compare differences in online classes and screen time between grades, and multinomial Logistic regression was performed to analyze the correlates of recreational screen time.@*Results@#The prevalence of prolonged recreational screen time was 42.2% and 55.2% on weekdays and weekends, respectively. Recreational screen time increased by 40.31 min/d on weekdays and 33.07 min/d on weekends, compared to usual school semesters. The average duration of an online class was (26.07±9.62) min, which totaled (110.41±51.98)min per day. Sex, grade, being the only child, and parents education levels were identified as the influencing factors of prolonged recreational screen time. Children who practiced moderate levels (weekdays: OR =1.27; weekends: OR =1.40; P <0.05) or lower levels of physical activity (weekdays: OR =1.86; weekend: OR =1.84; P < 0.05 ) were at a higher risk of prolonged recreational screen time than those who practiced more vigorous physical activity. Children whose parents limited their screen time to a moderate (weekdays: OR=1.61, P <0.05) or lower level (weekdays: OR=1.32, P < 0.05 ) had a higher risk of prolonged recreational screen time than those with a higher frequency. Children with parents recreational screen time ≥ 2 h/d had a higher risk of prolonged recreational screen time than the reference group; children who exhibited moderate to vigorous levels of physical activity <1 h/d (weekdays: OR=1.31, P <0.05), and those used electronic devices for learning 1-2 h/d (weekdays: OR =2.65; weekend: OR =2.65; P <0.05) or for ≥2 h/d (weekdays: OR =4.05, weekend: OR=5.24, P < 0.05 ) were at a higher risk of prolonged recreational screen time than the reference group.@*Conclusion@#During the COVID-19 pandemic, the level of screen time among children in Guangdong was high. Behavioral monitoring and targeted interventions are needed to promote children s health.

Emotional and behavioral problems and determinants among primary and middle school students aged 6 to 17year-old in Guangdong Province during the COVID-19 pandemic / 中国学校卫生

Objective@#To gain a better understanding of the emotional and behavioral problems associated determinants of primary and middle school students from Guangdong Province during the COVID-19 pandemic, so as to provide a basis for developing targeted intervention strategies.@*Methods@#Using the method of convenience sampling, an online questionnaire survey was carried out among primary and middle school students from Guangzhou, Zhanjiang and Zhongshan from March to April 2020. The emotional and behavioral problems of primary and middle school students were assessed using the Conners Parental Symptoms Questionnaire(PSQ), and a self compiled questionnaire was used to collect basic information related to the primary and middle school students and the influencing factors of emotional and behavioral problems. A total of 7 755 valid questionnaires were retrieved and statistically analyzed using the chi square test and multivariate Logistic regression.@*Results@#The detection rate of emotional and behavioral problems among children and adolescents aged 6 to 17-years-old in Guangdong Province was 14.8%; that was 21.0%, 14.4%, 7.3 %, and 10.1% in lower primary school students, upper primary school students, junior high school students, and high school students, respectively. The detection rate of the psychosomatic and hyperactivity index in boys was higher than that observed in girls, and the detection rate of anxiety in boys was lower than that observed in girls( P <0.05). There were statistically significant differences in emotional and behavioral problems in children in different grades( P <0.05). The results of the regression analysis showed that male students were at risk of psychosomatic ( OR= 1.37 , 95%CI =1.04-1.82) and hyperactivity disorders( OR=1.58, 95%CI =1.21-2.06), whereas the male gender was a protective factor for anxiety( OR=0.50, 95%CI =0.39-0.64). Grades were identified as the influencing factors of all of the factors related to emotional and behavioral problems. Students who reported excessive screen time and insufficient sleep were more likely to experience emotional and behavioral problems.@*Conclusion@#The detection rate of emotional and behavioral problems among primary and middle school students in Guangdong Province during the COVID-19 epidemic was high, which was associated with sex, grade, screen time and sleep. It is necessary to develop and implement targeted intervention measures.

Relationship between iodine nutrition level and iodine content in drinking water in Weinan City, Shaanxi Province / 中华地方病学杂志

Objective:To explore the relationship between iodine nutrition level and iodine content in drinking water in Weinan City, Shaanxi Province.Methods:In 2017, in 10 counties (cities, districts) under the jurisdiction of Weinan City, one township (town, sub-district office, hereinafter referred to as the township) was selected in each county (city, district) according to the 5 directions of east, west, south, north, and middle. One primary school was selected in each township, and 42 children aged 8 to 10 (age and gender balanced) in each primary school were selected, urine samples and household edible salt samples were collected to test the contents of urinary iodine and salt iodine, and children's goiter was examined. Twenty-one pregnant women were selected from each township, urine samples and household edible salt samples were collected to test the contents of urinary iodine and salt iodine. At the same time, the drinking water samples of residents in townships were collected to test water iodine content. The correlation between urinary iodine, water iodine and salt iodine was analyzed by multiple linear regression.Results:Urine samples of 2 100 and 1 050 were collected from children and pregnant women, and the median urinary iodine was 254.51 and 172.55 μg/L, respectively. Edible salt samples of 2 100 and 1 050 were collected from children and pregnant women, and the median salt iodine was 24.00 and 24.44 mg/kg, respectively. A total of 232 water samples were collected, and the median water iodine was 26.53 μg/L. A total of 2 100 children's thyroids were examined, of which 65 had goiters, and the goiter rate was 3.10%. The regression model of urinary iodine and water iodine in children was statistically significant ( F = 6.48, P < 0.05), the multiple linear regression equation was = 235.52 + 1.01 x, coefficient of determination ( R2) = 0.119, 11.9% of the change of urinary iodine was related to the change of water iodine. Conclusions:Children's iodine nutrition in Weinan City is at an ultra-adequate level, while pregnant women's iodine nutrition is at an appropriate level. Eleven point nine percent of the changes in children's urinary iodine can be explained by the changes in water iodine.

Effects of T-2 toxin on expression of hepatocyte growth factor and its receptor in articular cartilage and epiphyseal cartilage of rats under selenium deficiency / 中华地方病学杂志

Objective:To observe the effects of T-2 toxin on expression of hepatocyte growth factor (HGF) and HGF receptor (C-Met) in articular cartilage and epiphyseal cartilage of rats under low selenium condition.Methods:Twenty-four healthy male SD rats weighted 60-80 g were randomly divided into conventional diet group (selenium content of 101.5 μg/kg) and low-selenium diet group (selenium content of 1.1 μg/kg), with 12 rats in each group. After 30 days of feeding, the conventional diet group was further divided into conventional group and T-2 toxin group (100 μg·kg -1·d -1), and the low-selenium diet group was further divided into low-selenium group and low-selenium+T-2 toxin group (100 μg·kg -1·d -1), with 6 rats in each group. After 30 days of feeding, the rats were sacrificed and the cartilage of knee joint was taken, the morphological changes of knee articular cartilage and epiphyseal cartilage were observed by HE staining under light microscope. Immunohistochemical method was used to detect the expression of HGF and C-Met in knee articular cartilage and epiphyseal cartilage, and positive expression rates of HGF and C-Met were calculated. Results:Under light microscope, chondrocytes of articular cartilage and epiphyseal cartilage in low-selenium+T-2 toxin group were sparse, and the necrosis and structural area were found in the deep layer, and the extracellular matrix of chondrocytes in the region was degraded and light stained, and proliferating granulation tissue was visible nearby. The positive expression rates of HGF in articular cartilage [(21.97 ± 6.90)%, (49.41 ± 8.24)%, (76.39 ± 5.88)%] and epiphyseal cartilage [(23.36 ± 12.49)%, (58.43 ± 14.48)%, (66.59 ± 10.83)%] of rats in low-selenium, T-2 toxin and low-selenium+T-2 toxin groups were higher than those in conventional group [(9.13 ± 6.01)%, (11.14 ± 4.67)%, P < 0.05]. The positive expression rates of C-Met in articular cartilage [(25.34 ± 7.53)%, (58.21 ± 12.54)%, (81.46 ± 7.89)%] and epiphyseal cartilage [(35.21 ± 4.71)%, (40.84 ± 2.03)%, (49.41 ± 6.29)%] of rats in low-selenium, T-2 toxin and low-selenium+T-2 toxin groups were higher than those in conventional group [(11.21 ± 5.11)%, (12.12 ± 4.71)%, P < 0.05]. Conclusion:T-2 toxin may affect the expression of HGF and C-Met in articular cartilage and epiphyseal cartilage of rats under low selenium condition.

Effects of T-2 toxin on expression of fibroblast growth factor 8 and fibroblast growth factor receptor 3 in articular cartilage and bone marrow of rats under low selenium condition / 中华地方病学杂志

Objective:To study the effects of T-2 toxin on expression of fibroblast growth factor 8 (FGF8) and fibroblast growth factor receptor 3 (FGFR3) in articular cartilage and subchondral marrow of rats under low selenium condition, and to explore the mechanism of deep cartilage injury and secondary complications in Kaschin-Beck disease (KBD).Methods:Twenty-four healthy male SD rats weighted 60 - 80 g were selected, they were divided into conventional feed group (selenium content of 101.5 μg/kg) and low-selenium feed group (selenium content of 1.1 μg/kg) by random number table method, with 12 rats in each group. After 30 days of feeding, the conventional feed group was further divided into control group and T-2 toxin group (100 μg·kg -1·d -1), and the low-selenium feed group was further divided into low-selenium group and low-selenium+ T-2 toxin group, with 6 rats in each group. After 30 days of feeding, the rats were sacrificed and the knee cartilage with cancellous bone was taken. Pathological changes of knee cartilage were observed by HE staining. Immunohistochemical method was used to detect the expression of FGF8 and FGFR3 in cartilage and subchondral marrow of knee joint, positive expression rates of FGF8 and FGFR3 in articular cartilage were calculated, and the integrated optical density (IOD) values of FGF8 and FGFR3 positive expression in subchondral marrow were analyzed by Image-Pro Plus 6.0 software. Results:Under light microscope, chondrocytes in low-selenium+ T-2 toxin group were sparse, and empty chondrocytes in the deep and middle layers of articular cartilage increased, and chondrocytes died and became red cell shadows. The extracellular matrix dissolved and was slightly stained in deep region, turning into necrotic and unstructurized areas. Proliferating granulation tissue was visible nearby. The positive expression rate of FGF8 in articular cartilage of rats in low-selenium+ T-2 toxin group [(88.61 ± 10.97)%] was higher than that in control, low-selenium and T-2 toxin groups [(10.35 ± 2.48)%, (19.26 ± 3.08)%, (58.89 ± 9.29)%, P < 0.05]; IOD value of FGF8 positive expression in subchondral marrow [(16.73 ± 1.72) × 10 6] was higher than that in control, low-selenium and T-2 toxin groups [(1.20 ± 0.41) × 10 6, (4.33 ± 0.97) × 10 6, (12.80 ± 1.12) × 10 6, P < 0.05]. The positive expression rate of FGFR3 in articular cartilage of rats in low-selenium+ T-2 toxin group [(89.76 ± 8.59)%] was higher than that in control, low-selenium and T-2 toxin groups [(13.18 ± 2.25)%, (21.15 ± 2.33)%, (32.55 ± 6.72)%, P < 0.05]; IOD value of FGFR3 positive expression in subchondral marrow [(16.50 ± 5.36) × 10 6] was higher than that in control, low-selenium and T-2 toxin groups [(7.58 ± 1.02) × 10 6, (10.73 ± 7.13) × 10 6, (9.83 ± 5.63) × 10 6, P < 0.05]. Conclusion:Under low selenium condition, T-2 toxin changes expression of FGF8 and FGFR3 in deep chondrocytes of articular cartilage and subchondral marrow in rats, elevated expression of FGF8 and FGFR3 may be involved in the occurrence and development of secondary changes in KBD.

Relationship between insulin-like growth factor-1 receptor down-regulation and Kaschin-Beck disease / 中华地方病学杂志

Objective To observe the expression level of insulin-like growth factor-1 receptor (IGF-1R) in the cartilage tissue of children with Kaschin-Beck disease (KBD) and T-2 toxin-poisoned rats under low selenium condition,and the effect of IGF-1R inhibitor on apoptosis of human normal chondrocytes (C28/I2 cells),and to investigate the role of IGF-1R in the pathogenesis of KBD.Methods The knuckles of dead children (5 cases) in the KBD areas,car accident death and congenital 6 finger deformity operation children (5 cases) in non-KBD areas in Shaanxi were collected,the expression of IGF-1R in the articular cartilage was detected by immunohistochemistry.Thirty-two male Sprague-Dawley rats with a body mass of 60-80 g were selected,according to the body mass,they were divided into the routine feed group (selenium content:101.5 μg/kg) and the low-selenium feed group (selenium content:1.1 μg/kg) by random number table method,16 rats in each group.After 30 days of feeding,the routine feed group was divided into control group and T-2 toxin group (100 ng·kg-1·d-1),the low-selenium feed group was divided into low selenium group and low selenium + T-2 toxin group,8 rats in each group,the expression of IGF-1R in the articular cartilage of the left knee joint was detected by immunohistochemistry after 30 days of feeding.C28/I2 cells were cultured in vitro and treated with T-2 toxin 0 (control),6,12,and 24 μg/L,and each concentration of T-2 toxin was accompanied with sodium selenite (+ 0.1 mg/L) for 72 h.Meanwhile,IGF-1R inhibitor with 0 (control),250,500,and 1 000 μg/L was treated on C28/I2 cells for 48 h.The expression levels of IGF-1R mRNA and protein in chondrocytes were detected by Real-time PCR and Western blotting,and the apoptosis of chondrocytes was detected by flow cytometry.Results Compared with the control group [(100.00 ± 0.00)％,(100.00 ± 0.00)％],the expression rates of IGF-1R positive cells in articular cartilage surface and middle layers [(72.71 ± 4.75)％,(36.33 ± 4.32)％] of children in KBD group were significantly reduced (t =12.852,32.650,P ＜ 0.01).Compared with control group [(100.00 ± 0.00)％,(100.00 ± 0.00)％,(100.00 ± 0.00)％],the expression rates of IGF-1R positive cells in articular cartilage middle layer [(20.83 ± 2.69)％,(26.45 ± 2.84)％,(20.34 ± 1.82)％],deep layer [(33.55 ± 5.66)％,(48.89 ± 8.39)％,(25.51 ± 7.50)％],and the expression rates of IGF-1R positive cells [(47.50 ± 1.47)％,(28.66 ± 3.58)％,(40.52 ± 6.78)％] in the hypertrophic layer of the metaphyseal plate of rats in low selenium,T-2 toxin,and low selenium + T-2 toxin groups were significantly reduced (P ＜ 0.01).C28/I2 cells were cultured in vitro,compared with the control group,IGF-1R mRNA and protein expression levels in each T-2 toxin groups were significantly reduced (P ＜ 0.05).The expression levels of IGF-1R mRNA (1.95 ± 0.35,2.44 ± 0.17,2.40 ± 0.15) in 6,12,24 μg/L T-2 toxin + 0.1 mg/L selenium groups were significantly higher than those in T-2 toxin groups (0.80 ± 0.08,0.63 ± 0.08,0.61 ± 0.11,t =-12.259,-11.279,-13.371,P＜ 0.05).The expression levels of IGF-1R protein (1.67 ± 0.70,1.07 ± 0.26) in 6,12 μg/L T-2 toxin + 0.1 mg/L selenium groups were significantly higher than those in T-2 toxin groups (0.52 ± 0.05,0.72 ± 0.05,t =-25.977,-10.776,P ＜ 0.05).Compared with the control group [(5.33 ± 0.85)％,(4.03 ± 1.15)％],C28/I2 cells early apoptosis rates [(8.26 ± 1.51)％,(13.00 ± 0.72)％,(13.19 ± 1.05)％] in each of IGF-1R inhibitor groups,and late apoptosis rates [(8.50 ± 0.71)％,(14.21 ± 1.10)％] in 500,1 000 μg/L IGF-1R inhibitor groups were increased significantly (P ＜ 0.05).Conclusions The expressions of IGF-1R in the cartilage tissue of KBD children and T-2 toxin-poisoned rats under low selenium condition are decreased.T-2 toxin decreases the expression of IGF-1R in chondrocytes,and selenium can partly inhibit the effect of T-2 toxin on IGF-1R.Down-regulation of IGF-1R causes chondrocyte apoptosis,and it may play an important role in KBD chondrocyte apoptosis.

Role of vascular cell adhesion molecule-1 in abnormal differentiation of chondrocytes in Kaschin-Beck disease / 中华地方病学杂志

Objective To investigate the expression of vascular cell adhesion molecule-1 (VC.AM-1) in oxidative stress induced hypertrophic chondrocytes,in Kaschin-Beck disease (KBD) patients and in rat fed with T-2 toxin under selenium deficient conditions in order to analyze the relationship between VCAM-1 biological function and the dysregulation of chondrocyte differentiation in KBD.Methods The ATDC5 was cultured in 1％ ITS solution (10 mg/L insulin,5.5 mg/L transferrin,and 6.7 μg/L sodium selenite) for 21 days,and stimulated with 3-morpholino-sydnonimine (SIN-1,a nitric oxide [NO] donor) to obtain the oxidative stress induced hypertrophic chondrocytes.Real-time PCR was used to detect VCAM-1 mRNA in hypertrophic chondrocytes induced by different concentrations of SIN-1.The expressions of VCAM-I in articular cartilage of child and adult KBD patients and KBD animal model were determined via the immunohistochemical method,and KBD cartilage samples were obtained in KBD areas from KBD child who had died or from adults who had had surgery.Results After treatment of hypertrophic chondrocytes (ATCD5 cells) with SIN-1 (0,1,3,5 mmol/L),VCAM-1 mRNA levels (1.00 + 0.00,1.22 ± 0.20,0.71 ± 0.22,0.37 ± 0.16) were decreased in a dose-dependent manner when compared with the control group (F =27.788,P ＜ 0.05).The densities of VCAM-1 positive cells in superficial and middle zones of the articular cartilage of children KBD patients [(16.08 ± 5.20)％,(19.20 ± 9.71)％] were higher than those of control group [(0.00 ± 0.00)％,(0.00 ± 0.00)％],while that in the deep zone [(7.00 ± 4.40)％] in children KBD patients was significantly lower than that of control [(51.60 ± 20.58)％,tS/M/D=-10.972,-6.249,6.564,P ＜ 0.05].The positive cell density of VCAM-1 in the adult patients was significantly increased in the superficial zone [(7.92 ± 4.29)％ vs (3.12 ± 1.12)％] but significantly decreased in the middle zone [(17.54 ± 8.27)％ vs (31.75 ± 13.30)％] of articular cartilage when compared with that of control group (tS/D =-3.824,3.037,P ＜ 0.05).In articular cartilage of the four groups of KBD rats,the density of VCAM-1 positive cells in the superficial zone was significantly higher in low selenium diet group,T-2 toxin diet group and selenium deficient plus T-2 toxin diet group [(4.11 ± 1.90)％,(5.00 ±2.02)％,(2.78 ± 1.48)％ vs (1.89 ± 1.76)％,P ＜ 0.05].But the density of VCAM-1 positive cells in the deep zone was significantly lower in rat feed with selenium diet and selenium deficient plus T-2 toxin diet [(13.67 ± 2.45)％,(20.56 ± 7.42)％] than that of control group [(33.00 ± 12.57)％,P ＜ 0.05] in the epiphyseal cartilage of KBD rats.Conclusions The level of VCAM-1 is decreased both in the SIN-1 induced hypertrophic chondrocytes and in the deep zone of articular cartilage in KBD patients and in rat fed with T-2 toxin and selenium-deficient diets.VCAM-1 may be associated with the death of deep zone chondrocytes and differentiation disorder in cartilage.

Clinical Study on Heat Shock Protein 70 in Skin Tissue of Patients with Pemphigus by Immunohistochemistry / 现代检验医学杂志

Objective This study was performed to investigate the levels of HSP70 in tissue in pemphigus as a possible new theoretical basis for further elucidate the pathogenesis of pemphigus.Methods The expression of HSP70 in 62 patients with pemphigus was determined by immunohistochemistry,and the normal skin was taken as control.Results The results showed that the positive cells of HSP70＞75 ％ in the blisters of pemphigus vulgaris and the positive cells of HSP70＞50％ in the inflammatory cells near the blisters,and the expression of HSP70 was significantly higher than that in normal skin,which was statistically significant(Z=5.42,4.73,P＜0.01).Conclusion The abnormal expression of HSP70 in inflammatory cells and psoriasis of pemphigus patients showed that HSP70 is involved in the pemphigus.

Apoptosis and necroptosis in articular cartilage in rat induced by T-2 toxin under selenium deficient conditions / 中华地方病学杂志

Objective To investigate the death of chondrocytes in rats which feed with T-2 toxin under selenium (Se) deficient conditions.Methods Thirty two healthy male SD rats were divided into two groups by weight which were normal diet group and Se deficiency diet group,16 rats in each group.Rats in normal diet group were fed with Se 101.5 μg/kg diet,and rats in Se deficiency diet group were fed with Se 1.1 μg/kg diet for 30 d.Normal diet group was divided into control group and T-2 toxin group,and Se deliciency diet group was randomly divided into Se-deficiency group and Se-deficiency plus T-2 toxin group,8 rats in each group.After that,rats in T-2 toxin and Se-deficiency plus T-2 toxin groups were administrated intragastrically with T-2 toxin (100 μg/kg) everyday for 30 d.Rats were put to death,the left knee was taken and stained with hematoxylin-eosin and SafraninFast green,pathological changes of rat's knee joint cartilage were observed under light microscopy,expression levels of active caspase-3 and receptor interacting protein 3 (RIP3) in rat's articular cartilage cells were determined via the immunohistochemical method.The apoptosis was also detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL).Results Red ghost outlines of chondrocyte and multiple chondral cell clusters surrounded the non-cell areas in deep zone of articular cartilage of knee joint stained with hematoxylineosin were seen in Se-deficiency plus T-2 toxin group under light microscope.In the superficial zone of cartilage,the positive percent of TUNEL and active caspase-3 in Se-deficiency plus T-2 toxin group was higher than those of control group,Se-deficiency group and T-2 toxin group [(7.47-± 0.34)％ vs (4.68 ± 0.54)％,(2.67-± 0.64)％,(2.56 ±0.54)％;(4.75 ± 0.67)％ vs (1.24 ± 0.25)％,(0.00 ± 0.00)％,(0.00 ± 0.00)％,P ＜ 0.05].In the middle zone of cartilage,the positive percent of TUNEL,active caspase-3 and RIP3 in Se-deficiency plus T-2 toxin group was significantly higher than those of control group,T-2 toxin group and Se-deficiency group [(72.06 ± 6.15)％ vs (16.10 ± 3.00)％,(19.57 ± 3.49)％,(19.33 ± 5.19)％;(51.13 ± 4.18)％ vs (10.97-± 3.01)％,(15.36 ± 4.37)％,(15.23 ± 3.13)％;(25.91 ± 13.39)％ vs (1.59 ± 1.14)％,(4.32 ± 2.91)％,(7.50 ± 5.00)％,P ＜ 0.05].The positive percents of TUNEL,active caspase-3 and RIP3 were not significantly different in the deep zone (P ＞ 0.05).Conclusion The death of the middle zone in the rat cartilage induced by T-2 toxin under selenium deficient conditions isapoptosis and necroptosis.

Oxidative damage induced by T-2 toxin toxication in articular cartilage of rats under selenium deficiency / 中华地方病学杂志

Objective To study the change of rats serum malondialdehyde (MDA) and the expression levels of 4-hydroxy acid nonene (4-HNE) and 8-hydroxy uridine (8-OHdG) of articular cartilage under low selenium (Se) and T-2 toxin poisoning,to explore oxidative damage of articular cartilage in rats.Methods Thirtytwo healthy male SD rats were divided into two groups by weight which were normal diet group and Se-deficiency group,16 rats in each group.Rats in normal diet group was fed with selenium 101.5 μg/kg diet,and rats in Sedeficiency group was fed with selenium 1.1 μg/kg diet for 30 d.Normal diet group was divided into control group and T-2 toxin group,and low selenium diet group was randomly divided into Se-deficiency group and Se-deficiency plus T-2 toxin group,8 rats in each group.After that,rats in T-2 toxin and Se-deficiency plus T-2 toxin groups were administrated intragastrically with T-2 toxin (100 mg/kg) everyday for 30 d.Rats were put to death,the left knee was taken and stained with hematoxylin-eosin and Safranin-Fast green,pathological changes of rat's knee joint cartilage were observed under light microscopy,expression levels of 8-OHdG and 4-HNE in rat's articular cartilage cells were determined by immunohistochemical method and rat's MDA content was determined by glucosinolates barbituric acid method.Results Chondronecrosis in deep zone of articular cartilage of knee joint stained with hematoxylin-eosin was seen in Se-deficient plus T-2 toxin diet group under light microscope.Significantly less Safranin-Fast green staining was observed in the cartilage of knee joints in the Se-deficient plus T-2 toxin diet group compared to the control group.Compared with control group [(3.41 ± 2.48)％,(2.28 ± 1.74)％],8-OHdG and 4-HNE in Se-deficient plus T-2 toxin group [(62.61 + 10.97)％,(75.03 ± 7.92) ％] positive expression rate increased significantly (F =16.24,18.61,all P ＜ 0.05).Comparison of serum MDA content in each group,the difference was statistically significant (F =4.32,P ＜ 0.05).The Se-deficiency group [(2.803 ± 0.163) μmol/L] was compared with control group [(1.873 ± 0.475) μmol/L] that the contents of serum MDA were increased.The T-2 toxin group [(2.890 ± 0.453) μmol/L] was compared with control group [(1.873 ± 0.475) μmol/L] that the content of serum MDA was increased (P ＜ 0.05).The Se-deficiency plus T-2 toxin group [(3.521 ± 0.292) μmol/L] was compared with Sedeficiency group and control group that the contents of serum MDA were increased (all P ＜ 0.05).Conclusions The marker of peroxidation products are increased in articular cartilage of SD rats under the condition of Sedeficiency and T-2 toxin poisoning.The cartilage damage and chondronecrosis due to Se-deficiency and T-2 toxin poisoning are related to oxidative damage.

Expressions of interleukin-1β,interleukin-6 and tumor necrosis factor alpha in articular cartilage of children with Kashin-Beck disease / 中华地方病学杂志

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.

Correlation of matrix metalloproteinases and Kashin-Beck disease / 中华地方病学杂志

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.

Clinical validation of a domestic fibrinogen reagent on coagulometer Sysmex CA1500 / 国际检验医学杂志

Objective To validate the clinical performance of a domestic fibrinogen reagent by freezing method(Clauss method) on the coagulometer Japan Sysmex CA1500.Methods The domestic fibrinogen reagent as the reagent A and the Germany Siemens Dade thrombin reagent as the reagent D,the Clauss method was adopted to measure the within-run precision and between-run preci-sion in two levels of quality control respectively.The reference value range was verified by the reagent A in 165 cases of normal clin-ical samples.The fibrinogen detection results in 200 cases of clinical samples were compared between by the reagent A and the rea-gent D.The significance test and the equivalence test were performed.Results The within-run precision CV of the reagent A and D in two levels of quality control were 4.28%,6.98% and 3.45%,5.22% respectively,the between-run precision CV of the reagent A and D in two levels of quality control were 6.23%,10.34% and 6.20%,9.89% respectively,the differences had no statistical significance(P >0.05).The reference value range of the reagent A was 2.08 -3.92 g/L.The fibrinogen detection results of the clinical samples by the reagent A and D had the statistically significant differences (P =0.025).But the 90% bilateral confidence in-terval(90%CI :-0.09,0.15)of the difference in the mean detection results by these two reagents located in the equivalent interval (-0.27,0.27).Conclusion The domestic fibrinogen reagent for Clauss method has reliable detection results and is suitable for the coagulometer Japan Sysmex CA1500,which is equivalent to the clinical application of Germany Siemens Dade thrombin reagent.

Research on matrix proteoglycan turnover of tissue engineering repaired cartilage / 西安交通大学学报(医学版)

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.

DETECTION OF HPVB19 IN SERA OF KASHIN-BECK DISEASE PATIENTS / 药物分析学报

Objective To investigate the relationship between Kashin-Beck Disease(KBD) and Human Parvovirus B19(HPVB19).Methods HPVB19DNA was detected in 55 sera of KBD patients and 52 healthy in adjacent non-endemic area and 35 healthy sera in normal area using PCR and then linked the HPVB19DNA to pGEM-T vector.The nucleotide sequence was analyzed and compared with HPVB19 nucleotide sequence published by Genebank and another in Journal of virology.Results HPVB19DNA was found in 16 of 55 sera in KBD patients,and the HPVB19DNA position rate(29.09%) is significantly higher than that of the two healthy control groups(11.54%、11.42% respectively)(P＜0.05).The nucleotide sequence homologies compared with the two published nucleotide sequence were 97.75%、97%,respectively.The putative amino acid homologies compared with the tow published were 93.5%.The amino acid variation was greater than the nucleotide sequence variation because of a base insertion.Conclusion There was a close relationship between HPVB19 infection and Kashin-Beck Disease.