1.Effect and mechanism of bone marrow-derived mesenchymal stem cells transplantation on collagen-induced arthritis
Chinese Journal of Rheumatology 2009;13(6):-
ObjectiveTo investigate the effect of bone marrow-derived mesenchymal stem cells (MSCs) on collagen-induced arthritis(CIA) and the possible mechanisms. MethodsMSCs from C57BL/6 mice bone marrow were isolated and expanded. Three DBA-I mice were as normal control group (Sodium Chloride injected on day 0 and 21) after primary immunization. After the bovine collagen Ⅱ induced-CIA models were established, fifteen male mice were randomly divided into 3 groups according to treatment regi-mens: CIA control group(0.2 ml Sodium Chloride injected through caudal vein on day 0 and 21), prevention group(1×10+6 cells MSC on day 0) and treatment group. (1×10+6 cells MSC on day 21 after primary immun-ization). The observed parameters included arthritis index (Al), articular pathology changes, levels of serum TNF-α, IL-1β detected hy ELISA. In addition, the percentage of CD4+CD25+Foxp3+Treg cells(Tregs) in CD4+ T cells of spleen or lymphoid node from mice were also analyzed by flow cytometry(FCM). Results①The Al and articular pathological score of the prevention group and treatment group were lower than the CIA control group(P<0.05). ②The levels of serum TNF-α and IL-1β of normal control, prevention group and treatment group were significantly decreased compared with CIA control group(P<0.05), and positively correlated with Al or articular pathological score (P<0.05). ③The percentage of Tregs from spleen and lymph nodes was significantly increased in MSCs treated mice(P<0.05). Conclusion MSCs transplantation is effective for CIA and suppression of TNF-α, IL-1β and up-regulation of Tregs by MSCs may be the possible mechanism in CIA.
2.Effect of OAZ signaling blocking through small interfering RNA on the production of anti-nuclear antibody in systemic lupus erythematosus
Rongliang LI ; Jin HUANG ; Tao ZHOU ; Lingyun SUN ; Xuebing FENG
Chinese Journal of Rheumatology 2010;14(1):3-7
Objective To explore the role of OAZ gene in the pathogenesis of systemic lupus erythe-matosus (SLE) by using RNA interfering technique. Methods Peripheral blood mononuclear cells (PBMC) from SLE patients were collected. Each sample was equally divided into four groups for cell culture in 96 well plates. Specific siRNA for OAZ and GAPDH were concordantly added to the experimental group and the positive control group, while nonspecific siRNA was added to the negative control group and only culture medium was added to the Mock control group. Cells and supernatants were harvested after culturing for 72 hours, then RNA was extracted and reverse transcripted to cDNA. OAZ, Id1, Id2, Id3, Id4 mRNA expression levels were analyzed by using real-time PCR. Levels of IFN-γ, IL-4, IL-10, IL-12, IL-21, CCL2, ANA in the supernatant were tested by ELISA. Relationships between the expression levels of OAZ mRNA with levels of cytokines and ANA were analyzed. Results OAZ, Id1, Id2, Id3 gene mRNA expression levels (△Ct: 12.5±1.4, 8.9±1.5, 4.3±0.8, 8.04±1.1) in the experimental group were significantly decreased comparing to those in the negative control group (△Ct: 10.2±1.1, 6.5±1.2, 2.4±1.3, 6.2±1.2 respectively, P<0.05). Levels of IFN-γ, IL-10, IL-12, IL-21 and ANA in the experimental group were significantly lower than those in the negative control group (P< 0.05), but level of CCL2 was higher than the negative control group (P<0.05). Difference of OAZ mRNA expression levels (△△Ct) between the experimental group and the negative control group was negatively correlated with changes of ANA, IL-21 levels, but positively correlated with changes of Th1/Th2, CCL2. Conclusion OAZ siRNA can effectively reduce the expression of genes involved in the OAZ signaling pathway in SLE. OAZ may lead to abnormal production of ANA via regulating Id genes and cytokines.
3.Evaluation of the treatment effect of internal jugular vein intervention therapy in patients with cerebral venous sinus thrombosis with color Doppler ultrasound
Lingyun JIA ; Yang HUA ; Xunming JI ; Tao LUO ; Kaiyuan ZHANG
Chinese Journal of Cerebrovascular Diseases 2016;13(7):360-364
Objective To investigate the feasibility and effectiveness of internal jugular vein (IJV) intervention therapy in patients with cerebral venous sinus thrombosis (CVST)with color Doppler ultrasound (CDU). Methods Twelve patients with CVST diagnosed by CDU and the 13 IJV lesions (localized luminal stenosis in 9 cases,venous long-segment slender in 2 cases,and right IJV localized luminal stenosis,and long-segment slender on the left in 1 case)confirmed by magnetic resonance venography (MRV)and/ or digital subtraction angiography (DSA)were enrolled retrospectively. CDU examinations were used at 1 week before and after IJV intervention therapy,6 months,1 year,and 2 years. The changes of the maximum diameter and the maximum velocity (V max )of the IJV were compared. The success rate and the long-term efficacy of the intervention therapy were analyzed. Results One week after treatment,the CDU examinations showed that the diameter of IJV stenosis in 13 IJV were increased significantly compared with those before procedure (4. 7 ± 2. 1 mm vs. 2. 3 ± 1. 3 mm;t = 5. 325,P < 0. 01). The velocity of blood flow of IJV was improved compared with before procedure (localized stenosis in 10 IJV[50 ± 15 cm/ s vs. 87 ± 24 cm/ s];t = 6. 285,P < 0. 01). Six of the 12 patients were followed up for a mean of 18 ± 7 months, two patients had restenosis after balloon dilatation. Conclusions For CVST patients with IJV lesions,the preliminary observation has indicated that IJV intervention therapy may improve the lesion lumen and hemodynamics. However,the intervention therapy,especially after balloon dilatation,the incidence of restenosis is higher. CDU can be used as an objective evaluation means for the long-term efficacy of IJV stenosis.
4.Genetic Characteristics and Research Progress of Feline Coronavirus
Laboratory Animal and Comparative Medicine 2024;44(6):661-666
Feline coronavirus (FCoV) is classified into two biotypes: feline infectious peritonitis virus (FIPV) and feline enteric coronavirus (FECV). FIPV and FECV might evolve and mutate via genetic recombination and mutation, leading to novel subtypes and variants. This study examined the genomic structure and biological subtyping of FCoV, analyzed the infection characteristics of FIPV and FECV, and investigated the mechanisms of FECV transforming into FIPV. The findings revealed that while their genome structures were fundamentally similar, differences in their ability to efficiently infect monocytes/macrophages significantly influenced their pathogenicity and transmission characteristics, with FIPV exhibiting higher virulence. Moreover, the analysis of the open reading frames (ORF)3/7 as well as the N/S sequences of FIPV indicated that its non-structural proteins were associated with modulation of the host immune system. These proteins enabled immune evasion, leading to more severe disease. The genomic variability of FCoV constitutes an important foundation for studying the pathogenicity and epidemiology of FIPV and FECV, and offers references for virus detection and drug development.
5.In vivo and in vitro experiment on the effect of beer on the serum enzyme activity
Hongyan GU ; Ru TAO ; Jing ZHAI ; Lingyun SUN ; Tao WANG ; Suyun BAI
Chinese Journal of Tissue Engineering Research 2006;10(4):190-192
BACKGROUND: There have been no reports available about the direct effect of beer on serum enzyme activity.OBJECTIVE: To observe the changes of the activity of various serum enzymes of the subjects in the in vivo and in vitro experiments after drinking bear.DESIGN: An observational controlled experiment.SETTING: The Institute of Basic Medicine of Taishan Medical College.PARTICIPANTS: The experiment was performed at the Institute of Basic Medicine of Taishan Medical College between March 2005 and April 2005.We selected 17 college students, aged 19 to 35 years, from Taishan Medical College, including undergraduate students and graduate students. In formed consents were obtained from the subjects before the experiment was conducted.METHODS: ① In vivo experiment: 3 mL of venous blood was collected from the subjects 3 hours after the ordinary diet as the control. Then, the subjects drank beer at an amount of 4 mL/kg according to their body mass immediately. 3 mL of blood was collected respectively 15, 30, 45, 60, 90,120, 80 minutes later to measure the changes of the activity of alanine aminotransferase, aspartate aminotransferase, γ-glutamyl transpeptidase, al kaline phosphatase, creatine kinase , lactate dehydrogenase, diastase and lipase. ② In vitro experiment: 17 fresh serum samples were added into two test tubes separately with 0.5 mL of serum in each tube. 20 μL of normal saline was added to the tube of the control and 20 μL of beer was added into the test tube. The direct effect of beer on the activity of various enzymes was observed.MAIN OUTCOME MEASURES: The changes of the activity of alanine aminotransferase, aspartate aminotransferase, γ-glutamyl transpeptidase, al kaline phosphatase, creatine kinase , lactate dehydrogenase, diastase and lipase of the serum on the in vivo and in vitro experiment .RESULTS: Totally 17 students were involved and all the students entered the stage of result analysis with no loss in the midway. ① In vivo experiment: Beer significantly decreased the activity of serum aspartate aminotransferase (418.08 ±58.68,383.41 ±63.01)nkat/L, significantly in creased the activity of serum alkaline phosphatase, (367 8.57±436.25,396 2.96±400.91)nkat/L (x2=19.00-20.00,P < 0.01). The activity of other enzymes was all increased at different degrees. ② In vitro experiment: Beer inhibited the activity of various enzymes in vitro to a certain degree.CONCLUSION: Beer has some effect on enzyme activity in vivo and in vitro, thus affecting the body metabolism. Over drinking beer can affect health. In the routine detection of serum enzymes, attention should be given to avoid the interference caused by over drinking beer to make sure the experimental results are precise and reliable.
6.Effects of beer on enzyme activity related to fat metabolism of rats
Hongyan GU ; Ru TAO ; Jing ZHAI ; Lingyun SUN ; Tao WANG ; Suyun BO
Chinese Journal of Tissue Engineering Research 2005;9(47):159-161
BACKGROUND: Long-term excessive intake of beer might lead to change of intra-corporal tissue or activity of serum enzyme.OBJECTIVE: Observation on relations between intake of beer and fat synthesis of rats and activity of enzyme correlated with catabolism in rats.DESIGN: Matched observations randomly of animal experiments.SETTING: Basic Medical Institute of Taishan Medical College.MATERIALS: The experiments were completed in Basic Medical Institute of Taishan Medical College from December 2004 to February 2005. Totally 60 SD rats were selected and categorized into 6 groups with 10 rats in each group. The rats were perfused with 9 mL/kg, 18 mL/kg, 27 mL/kg, 36 mL/kg, and 45 mL/kg beer respectively according to their fat; The rats in control group were fed with water in stead of beer.METHODS: All rats in each groups were narcotized and executed after continuous feeding 1 week, biochemical analysis and enzyme assay were made respectively after blood samples were adopted, and liver, subcutaneous fat, mesentery fat tissue and gastrocnemius muscle were preserved.MAIN OUTCOME MEASURES: ① The level of fat, blood glucose, insulin and blood-lipid of rats in each group after feeding 1 week . ② The enzymatic activity of liver and fat tissue of rats in each group after feeding 1 week . ③ The activity of hormone sensitive lipase and lipoprotein lipase (LPL) of rats in each group after feeding 1 week.RESULTS: Totally 60 rats were channeled into result analysis without any loss. ① Comparison of the levels of fat and biochemical specifications of rats in each group after feeding 1 week: The contents of fat, serum free fatty acid (FFA), high-density of lipoprotein (LP) cholesterol, hepar triacylglycerol and liver cholesterol in beer 36 mL/kg group were higher than those in control groups (x2=19.44-20.01, P < 0.01). ② Comparison of the levels of the enzymatic activity of liver and fat tissue of rats in each group after feeding 1 week: The activities of liver, subcutaneous fat, and liver microsome, I.e. Triacylglycerol alternation protein, phosphatidyl phosphohydrolase, malic enzyme, glucose-6-phasphate dehydrogenase (G6PD) of mesentery tissue in beer 36 mL/kg group were higher than those in control groups (x2=15.02-16.00, P < 0.05). ③ The comparison on level of the activity of hormone sensitive lipase and lipoprotein lipase (LPL) of rats in each group after feeding 1 week: The activity of gastrocnemius muscle of hormone sensitive lipase in beer 36 mL/kg group were prominently lower than those of control groups (P < 0.01), but the activity of lipoprotein lipase (LPL) (P < 0.01) of subcutaneous fat were prominently higher than those in control groups (P < 0.01). The activities of lipoprotein lipase (LPL)of mesentery fat tissue, subcutaneous fat and gastrocnemius muscle tissue in beer 36 mL/kg group were prominently higher than those in other beer groups and control groups (x2=19.00-20.00, P < 0.01).CONCLUSION: The intake of a certain amount of beer (36 mL/kg) might promote the capability of liver in synthesis and the transport of triacylglycerol in rats. The acceleration of lipid synthesis and storage of fat tissue such as mesentery fat tissue and the increase of fat decomposition and mobilization in peripheral tissue such as muscular tissue and subcutaneous fat would finally lead to the increase of fat.
7.Research on the clinical value of urinary NTx,serum BAP and BMD in patients with bone metastasis from malignant tumors
Lingyun WANG ; Xiaofang LIU ; Tao DU ; Lijia LI ; Xiaoyan HU ; Lei LIU ; Ming LI
Chongqing Medicine 2014;(9):1061-1062,1065
Objective To evaluate the variance and clinical significance of urinary pyridinoline cross-linked N-telopeptides of Type Ⅰ collagen(uNTx) ,serum bone specific alkaline phosphates(sBAP) and Bone mineral density(BMD) in patients with bone metastasis malignant tumors .Methods The levels of 53 case patients′,who with bone metastasis ,uNTx and sBAP were measured by ELISA and BMD was detected by dual energy x-ray absorptiometer .40 cases of non-bone metastasis people was assayed .Results The levels of all bone markers in patients with bone metastasis was significantly higher than in the patients without bone metas-tasis(P<0 .05) .The levels of the two biomarkers showed a positive correlation with the number of metastatic loci in bone (P<0 .05) .But there was not significantly different between the levels of uNTx or sBAP with the severity of the bone pain (P>0 .05) . BMD decreased in patients with bone metastasis group ,but it was no difference in the non-bone metastasis(P>0 .05) .Conclusion uNTx and sBAP are helpful for early diagnosis and prevention of patients with bone metastasis from malignant tumors .Patients with malignant gumor is often accompanied by a decrease in BMD .
8.The expression of arginase Ⅰ in heptocellular carcinoma and its clinicopathological significance
Xuan TAO ; Bin WANG ; Aimin HUANG ; Yunbin YE ; Chuanzhong HUANG ; Lingyun GAO
Chinese Journal of Digestion 2012;32(2):113-117
Objective To investigate the expression of arginase Ⅰ(ARG1)in hepatocellular carcinoma(HCC)and to analyze its correlation with clinicopathological features.Methods The expression of ARG1 at protein level in 167 samples of HCC and corresponding adjacent liver tissue was detected with high-throughput tissue microarray technique and immunohistochemistry.The correlation between ARG1 expression and clinicopathological features was analyzed with x2 test and Spearman rank correlation analysis.The expression of ARG1 at mRNA level in 68 samples of HCC and corresponding adjacent liver tissue was determined by real-time polymerase chain reaction(real-time PCR).Results The expression of ARG1 at protein level in HCC(3.540±3.702)was significantly lower than that of the corresponding adjacent liver tissues(10.290 ± 2.303)(t=-22.421,P=0.000).The ARG1 expression was correlated with differentiation degree of HCC,histological grade,vascular invasion,preoperative level of α-fetoprotein(AFP)and recurrence after operation(all P<0.05).The ARG1 expression at mRNA level in 68 HCC tissue[0.0997(0.213)]was lower than that of the corresponding adjacent liver tissues[0.563(0.459)],and the difference was statistical significant(u=-6.544,P=0.000).Conclusion Low expression of ARG1 in HCC may take part inHCC genesis and development.Detecting the expression of ARG1 may be helpful in HCC diagnosis,differentiation degree and prognosis assessment.
9.The correlation of carotid artery structure characteristics and residual stenosis after stenting
Yunlu TAO ; Yang HUA ; Lingyun JIA ; Lili WANG ; Beibei LIU ; Chen LING
Chinese Journal of Ultrasonography 2017;26(6):490-493
Objective To analyze the correlation of characteristics of carotid artery structure and the incidence of residual stenosis after carotid artery stent (CAS) placement and its influencing factors using color Doppler flow imaging (CDFI).Methods Five hundred and ninety-six cases from January 2013 to December 2015 who underwent CAS (600 pieces of stent) were included in this study.All patients were examined by CDFI within 1 month before and 1 week after carotid artery stenting.The incidence of residual stenosis was analysed.The correlation of residual stenosis and the characteristics of carotid artery lesions and atherosclerotic plaque before stenting were analyzed respectively.Results There was positive correlation between the incidence rates of residual stenosis and irregularly shaped plaque (odd ratios,9.02;95% confidence interval,5.21-15.59,P<0.05),the plaques with calcification in the surface(odd ratios,2.55;95% confidence interval,1.45-4.49,P<0.05),the residual diameter of carotid stenosis less than 1.0 mm(odd ratios,1.61;95% confidence interval,1.06-2.45,P<0.05),which were the independent risk factors for influencing residual stenosis after CAS.Conclusions Choosing a more adaptable stent based on the characteristics of carotid artery lesions and atherosclerotic plaque by CDFI before stenting may be useful for the patients to get best result of revascularization.The rate of residual stenosis may be decreased.
10.Effect observation on S-1 joint high-frequency diathermic therapy and perfusion chemotherapy in the treatment of advanced gastric cancer complicated with seroperitoneum
Yi XIAO ; Xiaofang LIU ; Tao DU ; Lingyun WANG ; Jing MA ; Ping ZHU
Cancer Research and Clinic 2016;28(4):251-255
Objective To explore the therapeutic effect and safety of high-frequency diathermic therapy combined with oral S-1 and intraperitoneal perfusion chemotherapy on malignant seroperitoneum of advanced gastric carcinoma.Methods Fifty-two advanced gastric carcinoma patients with malignant seroperitoneum were divided into observation group and control group.Twenty-five patients in observation group were received DDP 60 mg/m2 combined with 5-Fu 600 mg/m2 and IL-2 3 000 000 U intraperitoneal perfusion chemotherapy,as well as oral S-1 of 40mg bid on day 1-14 of every 21 days for a total of two or three cycles.High-frequency diathermic therapy was administered thirty minutes after intraperitoneal perfusion chemotherapy,twice a week for 3 weeks.Twenty-seven patients in control group were treated with S-1 and intraperitoneal perfusion chemotherapy only.Results In observation group,4 patients were CR,15 patients were PR,5 patients were SD,1 patient was PD,and the response rate (CR+PR) was 76 %.In control group,3 patients were CR,14 patients were PR,7 patients were SD,3 patients were PD,and the response rate (CR+ PR) was 63 % (P > 0.05).Median survial time for observation group was 10.0 months,8.0 months for control group (P > 0.05).Karnofsky scores after treatment in observation group was higher than that in control group,patients' clinical benefit rate were 80.0 % (20/25) and 51.9 % (14/27).The difference in life quality between the two groups was statistically significant (P < 0.05).There was no significant difference in adverse reactions between the two groups (x2 =4.544,P =0.033).Conclusion High-frequency diathermic therapy combined with oral S-1 and intraperitoneal perfusion chemotherapy for advanced gastric carcinoma with malignant seroperitoneum has good short-term curative effects and security,and this method should be further studied.