BACKGROUND:Growth differentiation factor-5 can induce adipose-derived stem cels into chondrocytes in our previous studies, but it has not been reported that the adipose-derived stem cels induced by growth differentiation factor-5 can differentiate into chondrocytes on the type I colagen scaffold. OBJECTIVE:To investigate the chondrogenic differentiation ability of adipose-derived stem cels induced by growth differentiation factor-5 cultured on the type I colagen scaffold. METHODS:Adipose derived stem cels were isolated from rabbit adipose tissue, the cels morphology was observed using inverted phase contrast microscope and the phenotypes were identified using immunofluorescence. The exogenous growth differentiation factor-5 was added to the cultural media with the type I colagen scaffold so as to induce the chondrogenic differentiation. The cels morphology was observed using hematoxylin-eosin staining and scanning electron microscope after the induction by growth differentiation factor-5 for 14 days. Meanwhile, the type II colagen and aggrecan mRNA expressions of the induced cels were measured using RT-PCR after the induction by growth differentiation factor-5 for 7, 14, and 21 days.RESULTS AND CONCLUSION:The primary cultured adipose-derived stem cels proliferated adherently with the fusiform and polygonal distribution under inverted phase contrast microscope. The positive CD44, CD49d and negative CD106 were detected by immunofluorescence. The adipose-derived stem cels induced by growth differentiation factor-5 were wel adhered to the type I colagen scaffold and strongly proliferated. The large amounts of extracelular matrix existed on the surface of the induced cels under scanning electron microscope. RT-PCR agarose gel electrophoresis indicated that the type II colagen and aggrecan mRNA expressions of the adipose-derived stem cels induced by growth differentiation factor-5 with the type I colagen scaffold were significantly increased. Growth differentiation factor-5 can successfuly induce the chondrogenic differentiation of adipose-derived stem cels cultured on the type I colagen scaffold.

[ ABSTRACT] AIM: To study the protective effects of cannabinoid CB2 receptor agonist JWH133 on rat acute lung injury induced by paraquat (PQ).METHODS: Male Sprague-Dawley rats (n=72) were randomly divided into 4 groups.PQ group:PQ was administered intraperitoneally at the dose of 20 mg/kg;Low-dose JWH133 pretreatment group ( L-JWH133 group):JWH133 (5 mg/kg, ip) was administered 1 h before PQ exposure;high-dose JWH133 pretreatment group ( H-JWH133 group):JWH133 (20 mg/kg, ip) was administered 1 h before PQ exposure;control group:1 mL sa-line was administered intraperitoneally.Arterial blood, bronchoalveolar lavage fluid (BALF) and lung tissue samples were collected at 8 h, 1 d and 3 d after PQ exposure.PaO2 and the levels of TNF-αand IL-1βin BALF were measured via blood gas analyzer and ELISA, respectively.The pathological changes and lung injury scores were assessed at 3 d after PQ expo-sure.NF-κB and AP-1 protein levels were also determined by Western blotting.RESULTS:The decrease in PaO2 , struc-tural injury of the lung tissues, interstitial pulmonary edema, and the increase in IL-1βand TNF-αin BALF were observed in PQ-treated rats compared with control group.JWH133 pretreatment reduced the degree of lung tissue injury, decreased the levels of IL-1βand TNF-αin BALF and the NF-κB and AP-1 protein expression in the lung tissue compared with PQ group, especially in H-JWH133 group.CONCLUSION:CB2 receptor agonist JWH133 inhibits NF-κB and AP-1 protein expression in the lung tissues, and reduces the secretion of IL-1βand TNF-αin BALF after paraquat exposure, thus atten-uating paraquat-induced acute lung injury.

Objective To study inhibition effect of rosiglitazone on lung injury induced by paraquat. Methods 72 male SD rats were randomly divided into three groups ( n=24 ): model control group, paraquat ( PQ ) was administered intraperitoneally at the dose of 20 mg·kg-1;rosiglitazone group, rosiglitazone (10 mg·kg-1 , ip) was administered 1 h before PQ administration; blank control group, 1 mL 0. 9% sodium chloride solution was administered intraperitoneally. The concentration of TNF-α and IL-1β in serum was measured by ELISA at 4, 8 h and 1, 3 day(s) after PQ exposure. The lung injury scores and nuclear factor-kappa B( NF-κB) positive signal were investigated 3 days after PQ exposure by HE staining and immunohistochemistry, respectively. Protein expression levels of NF-κB and activating protein-1(AP-1) were also determined by using Western blotting. Results The levels of IL-1β and TNF-α in serum of PQ-treated rats were significantly increased as compared with blank control group. Rosiglitazone pretreatment reduced the degree of lung tissue injury, the levels of IL-1β and TNF-α in serum, and the protien expression levels of NF-κB and AP-1 as compared with the model control group. Conclusion Rosiglitazone can inhibit NF-κB and AP-1 protein expression in lung tissue, reduce the levels of IL-1β and TNF-α in serum after PQ exposure, and exert an inhibition effect on inflammation in PQ-induced lung injury of rats.

In this study the ability of the monoclonal anti-idiotypic antibody NP30 was tested as a substitute of diagnostic antigen in detecting antibody of Schistosoma japonicum from human sera by use of ELISA. The results showed that the seropositive rate was 98% with NP30 in the group of acute infection, which was comparable to 94% with gut associated antigens (GAA)and 98% with the soluble egg antigens (SEA); 87% with NP30 in the group of chronic infection which was comparable to 86% with GAA but lower than that of 98% with SEA. The false positive rate was about 3% for all three diagnostic antigens. The results also showed that the geometric mean titer (GMT) of antibody to NP30 was higher than that to GAA but lower than that to SEA in the acute infection group and the GMT of antibody to NP30 was lower than both those to GAA and to SEA in the chronic infection group,suggesting that the antibody to NP30 appeared earlier and decayed more quickly during the process of infection. The authors suggested that NP30 could be used for the diagnosis of schistosomiasis japonica.

AIM: To study the effects of taurine at different doses on renal oxidative stress and inflammation induced by paraquat in rats.METHODS: Male SD rats (n=48) were randomly divided into 4 groups: negative control group, paraquat group, paraquat + low-dose taurine group, and paraquat + high-dose taurine group.The serum levels of creatinine and urea nitrogen were detected by a biochemical analyzer.The levels of malondialdehyde (MDA) and superoxide dismutase (SOD) were measured by colorimetry.The plasma concentrations of IL-6 and intercellular adhesion molecule (ICAM)-1 were detected by ELISA.Renal reactive oxygen species (ROS) was checked by fluorescence probe dihydroethidium (DHE).The protein levels of renal p-P38 MAPK, p-ERK1/2 and p-JNK were determined by Western blot.The mRNA expression of TNF-α, IL-6 and TGF-β1 was detected by real-time PCR.RESULTS: Serum creatinine and urea nitrogen increased after paraquat poisoning, and decreased after feeding with taurine in poisoned rats, with better result in high-dose taurine group.Taurine reduced the oxidative stress and inflammation in the renal tissue, and also reduced the protein levels of p-JNK, p-ERK1/2 and p-P38 in the kidney of paraquat-poisoned rats.CONCLUSION: Taurine attenuates renal injury induced by paraquat poisoning in rats.The mechanism may be related to reducing renal MAPK activity, oxidative stress and inflammatory response.

Objective To improve the first-aid ability of emergency residents by using simulated clinical practice skills training combined with scenario analysis in standardized residency training. Methods 218 residents who participated in standardized residency training in Shengjing Hospital of China Medical University were randomly distributed to two groups. The experimental group (n=134) was trained with the teaching methods of simulated clinical practice skills training combined with scenario analysis; the control group (n=84) was trained with traditional teaching methods combined with teaching video. The theoretical test scores and clinical practice skills results of the two groups were compared, and a questionnaire survey was also conducted after comparing the two teaching modes. T test and rank-sum test were performed by using SPSS 19.0. Results There was no significant difference in the theoretical test scores between the experimental group (85.27 ±5.59) and the control group (84.11 ±7.43) (P>0.05). However, the clinical practice skills scores of the experimental group (89.42±6.63) were significantly higher than those of the control group (71.36±4.41) (P<0.05), which showed the statistical significance. Student's satisfactions with teachers and teaching methods in the experimental group were 70.90% and 80.60%, respectively, which were extremely higher than those in the control group (23.81% and 33.33%). Student's satisfaction with self-learning in the experimental group (64.93%) was significantly higher than that in the control group (39.29%). Conclusions The application of simulated clinical practice skills training combined with scenario analysis could improve the first-aid ability of emergency residents. This teaching method is expected to be applied in standardized residency training in emergency medicine in the future.

Objective:To investigate the predictive value of early indicators changes in blood test on the prognosis of patients with acute paraquat poisoning.Methods:The clinical data of patients with acute paraquat poisoning admitted to emergency department of Shengjing Hospital of China Medical University from January 2012 to June 2019 were retrospectively analyzed. The changes of blood test indexes within 24 hours after admission were collected, including white blood cell count (ΔWBC), neutrophils count (ΔNE), lymphocytes count (ΔLY), monocytes count (ΔMO), arterial partial pressure of oxygen (ΔPaO 2), arterial partial pressure of carbon dioxide (ΔPaCO 2), arterial blood pH (ΔpH), bicarbonate radical (ΔHCO 3-), base excess (ΔBE), lactate (ΔLac), total protein (ΔTP), albumin (ΔALB), alanine aminotransferase (ΔALT), aspartate aminotransferase (ΔAST), total bilirubin (ΔTBil), direct bilirubin (ΔDBil), blood urea nitrogen (ΔBUN), serum creatinine (ΔSCr), serum calcium concentration (ΔCa 2+), and serum potassium concentration (ΔK +). Multivariate Logistic regression was used to analyze the risk factors of prognosis in patients with acute paraquat poisoning, and receiver operating characteristic (ROC) curve was drawn to analyze the predictive value of ROC curve for the death of patients with paraquat poisoning. Results:A total of 251 patients with acute paraquat poisoning were included, with 99 cases dead, and the mortality was 39.4%. The increase of the markers including ΔWBC, ΔLac, ΔALT, ΔAST, ΔTBil, ΔDBil, ΔBUN, ΔSCr and ΔK + within 24 hours of admission in the death group were significantly higher than that in the survival group; the decrease of the markers including ΔPaCO 2, ΔHCO 3-, ΔBE, ΔTP, and ΔALB in the death group were significantly greater than those in the survival group. The variables with statistical significance in the above single factor analysis were included in the multivariate Logistic regression analysis. The results showed that ΔLac, ΔSCr and ΔK + were independent risk factors for the prognosis of patients with acute paraquat poisoning [odds ratio ( OR) and 95% confidence interval (95% CI) were 1.662 (0.997-2.772), 1.045 (1.010-1.083) and 4.555 (1.190-17.429), respectively, all P < 0.05]. The area under the ROC curve (AUC) of ΔLac, ΔSCr and ΔK + for predicting death of patients with acute paraquat poisoning was 0.639 (95% CI was 0.505-0773), 0.811 (95% CI was 0.704-0.917), and 0.649 (95% CI was 0.519-0.779), respectively. When the cut-off of ΔLac was 1.85 mmol/L, the sensitivity was 87.9%, the specificity was 47.7%, and the diagnostic accuracy was 70.2%; when the cut-off of ΔSCr was 37.75 μmol/L, the sensitivity was 84.4%, the specificity was 77.9%, and the diagnostic accuracy was 80.5%; when the cut-off of ΔK + was 0.42 mmol/L, the sensitivity was 36.6%, the specificity was 90.7%, and the diagnostic accuracy was 68.3%. The efficiency of combination of ΔLac, ΔSCr, and ΔK + was greater than a single indicator in predicting death of patients with acute paraquat poisoning, with AUC of 0.911, and 95% CI of 0.834-0.989. Conclusions:ΔLac, ΔSCr, ΔK + within 24 hours of admission were all independent risk factors for the prognosis of patients with acute paraquat poisoning. ΔSCr > 37.75 μmol/L within 24 hours of admission would predict a poor prognosis in the patients with acute paraquat poisoning. Combined analysis of ΔLac, ΔSCr, and ΔK + can predict the prognosis of paraquat poisoning patients more accurately than single index.

Objective:To investigate the effect of 125I-RSOAds-hTERT/PSA oncolytic adenovirus on targeted therapy of prostate cancer and its effect on tumor microenvironment. Methods:125I-RSOAds-hTERT/PSA ( 125I-virus complex) oncolytic adenovirus was constructed by PCR amplification and double restriction enzyme ligation. TUNEL staining, flow cytometry and Caspase-3 immunoblotting assay were used to detect the killing effect of 125I-RSOAds-hTERT/PSA oncolytic adenovirus on prostate cancer cells in vitro and in vivo, respectively. To explore the effect of 125I-virus complex on tumor tissue cytokine secretion levels, interleukin 2 (IL-2), IL-10, tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) in the culture supernatant of human prostate cancer cell line PC3, mouse prostate adenocarcinoma cell line RM-1, and mice serum were detected by ELISA. We explored the regulation of 125I-virus complex on the expression of CD24, CD44 and prostate stem cell antigen (PSCA) in prostate tumor tissues and tumor cells through immunohistochemistry. Meanwhile, the expression levels of CD32 and vascular endothelial growth factor (VEGF), as well as CD4+ , CD8+ and macrophage infiltration in tumor tissue were detected through immunofluorescence experiments. Results:125I-virus complex oncolytic adenovirus significantly increased tumor cell apoptosis in vitro and in vivo that was significantly higher than that of 125I group and virus complex group. Meanwhile, IL-2 ( t=-183.30, -38.20, P<0.05), IL-10 ( t=113.80, 92.71, P<0.05), TNF-α ( t=-73.20, -73.91, P<0.05), IFN-γ ( t=-65.37, -139.70, P<0.05) increased in vitro and in vivo. 125I-virus complex reduced the expression of CD24, CD44 and PSCA in tumor cells and tumor tissue, reduced the weight of tumor tissue, inhibited angiogenesis of tumor tissue ( t=8.55, P<0.05), and regulated the immune response in tumor tissue. Conclusions:125I-virus complex targeting prostate cancer can significantly kill cancer cells, reduce the weight and angiogenesis of tumor, and improve tumor microenvironment.

BACKGROUND:Lijin manipulation can promote skeletal muscle repair and treat skeletal muscle injury.However,the formation of fibrosis and scar tissue hyperplasia are closely related to the quality of skeletal muscle repair.To study the regulatory effect of Lijin manipulation on the formation of fibrosis and scar tissue hyperplasia is helpful to explain the related mechanism of Lijin manipulation to improve the repair quality of skeletal muscle injury. OBJECTIVE:To explore the mechanism of Lijin manipulation to improve the repair quality of skeletal muscle injury in rabbits,thereby providing a scientific basis for clinical treatment. METHODS:Forty-five healthy adult Japanese large-ear white rabbits were randomly divided into blank group,model group and Lijin group,with 15 rats in each group.Gastrocnemius strike modeling was performed in both model group and Lijin group.The Lijin group began to intervene with tendon manipulation on the 3rd day after modeling,once a day,and 15 minutes at a time.Five animals in each group were killed on the 7th,14th and 21st days after modeling.The morphology and inflammatory cell count of gastrocnemius were observed by hematoxylin-eosin staining,the collagen fiber amount was observed by Masson staining,the expression of interleukin-6 and interleukin-10 in gastrocnemius was detected by ELISA.The protein and mRNA expressions of paired cassette gene 7,myogenic differentiation factor,myoblastogenin,alpha-actin,transforming growth factor beta 1,and type Ⅰ collagen were detected by western blot and RT-PCR,respectively,and the expression of type Ⅰ collagen protein was detected by immunohistochemistry. RESULTS AND CONCLUSION:Hematoxylin-eosin staining and Masson staining showed that compared with the model group,inflammatory cell infiltration and collagen fiber content decreased in the Lijin group(P<0.01),and the muscle fibers gradually healed.ELISA results showed that compared with the model group,the expression of interleukin-6 in the Lijin group continued to decrease(P<0.05),and the expression of interleukin-10 increased on the 7th day after modeling(P<0.05)and then showed a decreasing trend(P<0.05).Western blot and RT-PCR results showed that compared with the model group,the protein and mRNA expressions of paired cassette gene 7,myogenic differentiation factor,myoblastogenin in the Lijin group were significantly increased on the 14th day after modeling(P<0.05),but decreased on the 21st day(P<0.05);the protein and mRNA expressions of alpha-actin,transforming growth factor beta 1,and type Ⅰ collagen in the Lijin group were significantly decreased compared with those in the model group(P<0.05).Immunohistochemical results showed that the expression of type Ⅰ collagen in the Lijin group was significantly lower than that in the model group(P<0.05).To conclude,Lijin manipulation could improve the repair quality of skeletal muscle injury by inhibiting inflammation,promoting the proliferation and differentiation of muscle satellite cells,and reducing fibrosis.