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OBJECTIVE: To study the effects of Tongbi Mixture 2, a compound traditional Chinese herbal medicine for treating rheumatoid arthritis (RA), on immunoregulation of T lymphocytes in rats with collagen-induced arthritis (CIA). METHODS: Forty Wistar rats were randomly divided into normal control group, untreated group, Tongbi Mixture 2-treated group, methotrexate (MTX)-treated group and Tripterygium wilfordii polyglycoside (TWP)-treated group. Except for the rats of the normal control group (injection with normal saline), rats of the other four groups were subcutaneouly multipoint-injected with collagen protein II to induce CIA. The rats were treated with normal saline, Tongbi Mixture 2, MTX tablets and TWP tablets respectively for 36 days. The expressions of CD28 and CD152 were detected by flow cytometry, while the content of tumor necrosis factor-alpha (TNF-alpha) was analyzed by enzyme-linked immunosorbent assay. RESULTS: The expression of CD28 among the three drug treated groups had no statistical difference (P>0.05), while significantly higher than that of the normal control group (P<0.01) and lower than that of the untreated group (P<0.01). The expression of CD152 in the Tongbi Mixture 2 treated-group was lower than those of the MTX- and TWP-treated groups (P<0.05, P<0.01), but had no statistical difference as compared with the normal control group (P>0.05). There were no statistical differences in content of TNF-alpha between the drug treated groups and the normal control group (P>0.05), but the content of TNF-alpha of the drug treated groups was lower than that of the untreated group (P<0.05 or P<0.01). CONCLUSION: Tongbi Mixture 2 can inhibit T lymphocytes through down-regulating the expressions of CD28 and CD152 and the content of TNF-alpha, which may be the major mechanisms in treating RA.

BACKGROUND: Researches on the pathogenesis and pathological changes of rheumatoid arthritis(RA) have achieved significant progress in recent years. But traditional Chinese medicine(TCM) has unique advantage in RA therapy.OBJECTIVE: To study effects of overall alkali in tongbiling(TBL) on the proliferation of lymphocytes and the transferrin receptor of T lymphocytes (CD71) to explore the mechanism of TBL on the modulation of cell immunity.DESIGN: A completely randomized grouping design and an explorative study by employing cells as subjects.SETTING: Sixth internal medicine department of a TCM university,center of tissue transplantation and immunology college of life science of a university PARTICIPANTS: The study was conducted in the central laboratory (tertiary laboratory of National TCM Administrator) of the first affiliated hospital of Guangzhou TCM medical university between July 2002 and August 2003. Ten clean male SD rats were selected.METHODS: Lymphocyte was separated from rat inguinal lymph node for culture. Concanavalin(ConA) was used for 72-hour stimulation. The impacts of overall alkali TBL on lymphocyte proliferation were tested by MTT.The expression of T lymphocyte CD71 was tested by flow cytometer after 48-hour stimulation of phorbol 12,13 -dibutyrate(PDB) or ConA.MAIN OUTCOME MEASURES: The impacts of overall alkali TBL on lymphocyte proliferation and T cell activation.RESULTS: Different concentration of overall alkali TBL could significantly inhibit the proliferation of lymphocytes under ConA stimulation. PDB and ConA-activated T lymphocyte CD71 + expressions were significantly higher than that of blank control group(P<0.01) . CD3+ CD71 + expressions [(62.03±1.51) %,(25.28±1.57) %,(20. 29±1.72)%] activated by ConA under different concentration of overall alkali TBL(50,100,200 mg/L)were significantly lower than(72.03±1.28)% of BPS-positive control group (P<0. 05). CD3 + CD71 + expressions activated by PDB under 100 mg/Land 200 mg/L of overall alkali TBL were significantly lower than that of phosphate buffer solution (PBS-)positive control group(P<0.05). Different concentration of overall alkali TBL had significant down-regulated effects on CD71 expression in T lymphocyte activated by PDB or ConA and there was also a significant dose-effect relationship(P<0. 05). The inhibition on ConA-activated CD71 expression was stronger than that of PDB.CONCLUSION: Overall alkali TBL can inhibit the abnormal proliferation of T lymphocyte and its mechanism might be realized through its inhibition on transferrin receptor.

[Objective] To observe the effects of Tongbiling (TBL) prescription in counteracting cartilage destruction in rats with rheumatoid arthritis (RA). [Methods] Collagen-induced arthritis (CIA) rat models were established to observe the therapeutic effects of TBL. After a short-term treatment, expressions of tumor necrosis factor (TNF - a) and transforming growth factor (TGF- P) in cartilage tissue of model group, methotrenxate (MTX) tablets group and Radix Tripterygii Wilfordii (RTW) preparation group were examined by immunohistochemical semi-quantitative assessment method. [ Results ] The number of chondrocytes with positive TNF - a expression was lower and the chondrocytes were stained lighter in normal group than those in model group and low-dose MTX group ( P

[ObjectivejTo explore the therapeutic mechanism of Tongbiling (TBL) prescription for rheumatoid arthritis (RA) by observing the effects of TBL on inflammation and expression of vascular endothelial growth factor (VEGF) in synovium of rats with collagen-induced arthritis (CIA) .[Methods]CIA rat models were established by subcutaneous injection of type II collagen. Pathological sections of synovium in knee joint were made after medication for 37 days and then stained by HE method and immunohistochemical method respectively. Infiltration of inflammatory cells and hyperplasia of macrophages A and fibrous tissue in HE-stained pathological sections were observed under microscope and expression of VEGF was observed by semi-quantitative immunohistochemical method. [ Results ] Infiltration of inflammatory cells and hyperplasia of macrophages A and fibrous tissue in high-dose TBL group were inferior to those in model group (P 0.05); both TBL and Radix Tripterygii Wilfordii at high dose decreased the expression of VEGF (P

[Objective]To observe the effects of total alkaloids (TA) of Tongbiling (TBL) prescription on proliferation of T lymphocytes and transferring receptor of T lymphocytes (CD71) in rats and to explore the mechanism of TBL in regulating cellular immunity. [Methods]Isolated lymphocytes from mesenteric lymph node were cultured with various concentrations of TBL TA and canavaline A (ConA) for 72 hours; the proliferation of T lymphocytes was observed by MTT method and CD71 expression rate of T lymphocyte activated by phorbol 12,13-dibutyrate (PDB) or ConA for 48 hours was detected by flow cytometry.[Results]Various concentrations of TBL TA inhibited the proliferation of T lymphocytes activated by ConA and decreased CD71 expression rate of T lymphocyte activated by ConA or PDB in a dose-effect manner. Inhibitory effect of TBL TA on CD71 expression rate of T lymphocyte activated by ConA was superior to that activated by PDB. [Conclusion]TBL TA can inhibit the abnormal proliferation of T lymphocytes and its mechanism may be related to the expression of CD71.

OBJECTIVE:To observe the efficacy and safety of Sanjiao fuzheng mixture combined with concurrent chemoradiotherapy of paclitaxel and nedaplatin in the treatment of local advanced non-small cell lung cancer (NSCLC). METHODS:A total of 68 patients with local advanced NSCLC selected from our hospital during Jan. 2015 to Jan. 2017 were divided into control group and observation group according to random number table,with 34 cases in each group. Control group was given Paclitaxel injection 135 mg/m2 intravenously,d1+Nedaplatin for injection 75 mg/m2 intravenously,d3,21 d as a treatment course,for 2 courses;routine fractionated intensity modulated radiation therapy,2 Gy each time,5 times a week,60-70 Gy in total;given 2 cycles of primary chemotherapy continuously after radiotherapy. Observation group was additionally given Sanjiao fuzheng mixture 250 mL/d,divided into 3 times,till the end of treatment,on the basis of control group. Clinical efficacies were observed in 2 groups. The levels of nutritional indexes (BMI,PAB,ALB,Hb) and tumor markers (SCC-Ag,CEA,TK1, CYFRA21-10) before and after treatment were observed. The occurrence of ADR were recorded. RESULTS:There was no statistical significance in the total effective rate between 2 groups (observation group 82.35％ vs. control group 73.53％)(P>0.05). After treatment,the levels of BMI,PAB,ALB and Hb in 2 groups were significantly lower than before treatment,but the observation group was significantly higher than the control group. The levels of SCC-Ag,CEA,TK1 and CYFRA21-1 in 2 groups were significantly lower than before treatment,and the observation group was significantly lower than the control group,with statistical significance (P<0.05). The incidence of Ⅲ-Ⅳ degree aleucocytosis,Ⅰ-Ⅱ degree hemoglobin reduction and thrombocytopenia in observation group were significantly lower than control group, with statistical significance (P<0.05). CONCLUSIONS:The efficacy of Sanjiao fuzheng mixture combined with concurrent chemoradiotherapy of paclitaxel and nedaplatin is similar to that of concurrent chemoradiotherapy of paclitaxel and nedaplatin for localadvanced NSCLC,which can improve nutritional status significantly,and reduce the incidence of ADR.

Objective To compare the etiological constitution of recurrent miscarriage (RM) between patients with consecutive two and three or more miscarriages through combining the routine examination results and embryonic karyotype. Methods Patients with a history of two or more consecutive clinical miscarriages(≤12 weeks of gestation)consulting in the RM clinic of the First Affiliated Hospital of Sun Yat-sen University from March 2011 to January 2016 were collected. Six hundred and ninety-six with detailed history recorded, routine clinical examinations of RM and at least once embryonic karyotype were ultimately enrolled in this study. Their etiological constitution of RM were analyzed in groups of consecutive two and three or more miscarriage. The etiologies of RM in analysis consisted of women age, body mass index (BMI), chromosome abnormalities of couples, uterine abnormalities, endocrinology abnormalities and antiphospholipid syndrome(APS). Results (1)Among 696 patients, the abnormal embryonic karyotypes was 60.6%(422/696)and routine RM etiologies was 32.2%(224/696), leaving the ratio of unexplained RM was only 29.0%(202/696).(2)A total of 717 embryo karyotype were found in 696 patients, included 21 cases with twice embryo karyotype results the percentage of normal embryo was 39.7%(285/717), while abnormal ones was 60.3%(432/717). Among the types of abnormal karyotype, the most common ones (>10%)were trisomy 16(19.2%, 83/432), monosome X(11.3%, 49/432)and trisomy 22(10.9%, 47/432). (3)Among the 696 RM patients, the number of two and three or more miscarriages were respectively 446(64.1%,446/696)and 250(35.9%,250/696). Comparing groups of three or more miscarriages with two miscarriages, there were significant differencein older age as well as uterine adhesion(P<0.05). But no difference was found in body mass index(BMI), the rates of chromosome abnormalities of couples, uterine abnormalities except uterine adhesion, endocrinology abnormalities and APS (all P>0.05) between two groups. Conclusions The abnormal embryonic karyotype is the most common cause of first-trimester RM. The etiological constitution of two and three or more recurrent miscarriages is accordant, suggesting that routine clinical examination and the embryonic karyotype should be started following two consecutive clinical early miscarriages.

Objective To investigate the value of automatic segmentation of carotid vessel wall in multicontrast MR images using U?Net neural network. Methods Patients were retrospectively collected from 2012 to 2015 in Carotid Atherosclerosis Risk Assessment (CARE II) study. All patients who recently suffered ischemic stroke and/or transient ischemic attack underwent identical, state?of?the?art multicontrast MRI technique. A total of 17 568 carotid vessel wall MR images from 658 subjects were included in this study after inclusion criteria and exclusion criteria. All MR images were analyzed using customized analysis platform (CASCADE). Randomly, 10 592 images were assigned into training dataset, 3 488 images were assigned into validating dataset and 3 488 images were assigned into test dataset according to a ratio of 6∶2∶2. Data augmentation was performed to avoid over fitting and improve the ability of model generalization. The fine?tuned U?Net model was utilized in the segmentation of carotid vessel wall in multicontrast MR images. The U?Net model was trained in the training dataset and validated in the validating dataset. To evaluate the accuracy of carotid vessel wall segmentation, the sensitivity, specificity and Dice coefficient were used in the testing dataset. In addition, the interclass correlation and the Bland?Altman analysis of max wall thickness and wall area were obtained to demonstrate the agreement of the U?Net segmentation and the manual segmentation. Results The sensitivity, specificity and Dice coefficient of the fine?tuned U?Net model achieved 0.878,0.986 and 0.858 in the test dataset, respectively. The interclass correlation (95% confidence interval) was 0.921 (0.915-0.925) for max wall thickness and 0.929 (0.924-0.933) for wall area. In the Bland?Altman analysis, the difference of max wall thickness was (0.037±0.316) mm and the difference of wall area was (1.182±4.953) mm2. The substantial agreement was observed between U?Net segmentation method and manual segmentation method. Conclusion Automatic segmentation of carotid vessel wall in multicontrast MR images can be achieved using fine?tuned U?Net neural network, which is trained and tested in the large scale dataset labeled by professional radiologists.

BACKGROUND:Deferoxamine mesylate is a potential anti-osteoporosis drug with iron chelation,vascular regeneration,and antioxidant effects.Recent studies have shown that the application of deferoxamine mesylate can be extended to the field of tissue regeneration engineering. OBJECTIVE:To investigate whether deferoxamine mesylate can promote the repair effect of iron overload osteoporotic rats after bone grafting for mandibular bone defects by simulating the state of iron accumulation in patients with postmenopausal osteoporosis with high iron intervention in osteoporotic rats. METHODS:An iron accumulation ovariectomized osteoporosis model was firstly constructed.The model group underwent bilateral ovariectomy,and the intraperitoneal injection of ferric ammonium citrate(90 mg/kg,twice a week,for 11 weeks)was started in the 2nd week,while the sham-operated group had some fat around the ovaries removed and was given an equal amount of saline for 11 weeks.After the successful modeling,the experimental rats were divided into sham-operated group(n=6),high iron ovariectomtized group(n=6)and high iron ovariectomized deferoxamine mesylate treatment group(deferoxamine mesylate group,n=6).Bone defects of 5 mm in diameter were established in the rat's bilateral mandibles and implanted with Bio-Oss bone powder.Intraperitoneal injection of deferoxamine mesylate(100 mg/kg,3 times a week)was started on postoperative day 4 in the deferoxamine mesylate group,and equal volume of saline was given in the sham-operated and high iron ovariectomized groups.The bone samples of the mandible,liver and blood were taken at 2 and 12 weeks after bone grafting for Prussian blue staining of the jaw and liver and ELISA detection of serum ferritin to detect iron levels in various body tissues;hematoxylin-eosin staining and Masson staining were performed to observe inflammatory cell infiltration and early osteogenesis in the bone defect area;tartrate resistant acid phosphatase staining was performed to observe osteoclast differentiation;ELISA was performed to detect serum calcitonin and type I collagen C-terminal peptide levels;and Micro-CT and hematoxylin-eosin staining were performed to observe osteogenesis in the middle and late stages. RESULTS AND CONCLUSION:The number of tibial trabeculae was reduced and the trabeculae were sparsely arranged in the high iron ovariectomized group.Iron levels in the liver,jaw bone and serum were significantly higher in the high iron ovariectomized group than the sham-operated group at 2 weeks after bone grafting,while the iron levels were significantly decreased after deferoxamine mesylate intervention(P＜0.05).In the early stage of bone defect repair,more inflammatory cell infiltration,less new bone matrix and less type I collagen fiber production were observed in the high iron ovariectomized group than in the sham-operated group(P＜0.05);after deferoxamine mesylate treatment,inflammatory cell infiltration was reduced,a small amount of new bone matrix was produced and collagen fibers increased significantly(P＜0.05).In the middle and late stages of bone defect repair,Micro-CT results showed a reduction in new bone production in the high iron ovariectomized group compared with the sham-operated group and increased new bone matrix after deferoxamine mesylate treatment(P＜0.05).Compared with the sham-operated group,the osteoclast number,serum calcitonin level,and serum type I collagen C-terminal peptide level were increased in the high-iron ovariectomized group,while the osteoclast number was decreased and bone metabolic indexes were improved after treatment with deferoxamine mesylate.To conclude,in ovariectomized rats with high iron intervention,elevated iron levels can be seen in multiple tissues,accompanied by reduced new bone production in the mandibular bone defect area.Deferoxamine mesylate can improve bone metabolism and inhibit osteoclast activity by removing iron deposits in tissues,improve bone formation in iron-accumulated osteoporotic rats,and promote bone healing in the mandibular bone defect area.