AIMTo study the effect of atropine, muscarinic cholinergic antagonist, on the central analgesic action of melatonin (MT) and to explore the mechanism of MT analgesia.

METHODSAs an indicator of visceral pain, the unit discharges of the neurons in the posterior group of thalamic nuclei (PO) were caused by stimulating the great splanchnic nerve (GSN) of the cat. The cranial stereotaxic and extracellular glass microelectrode record technique were used. The drugs were given through the intra-cranial-ventricle (icv).

RESULTS0.1% MT (10 micrograms.kg-1, icv) was shown to inhibit the unit discharge of the neurons in PO of the cat, whether the long latency or the short latency, which was evoked by stimulating GSN. The inhibition of 0.1% MT (10 micrograms.kg-1, icv) on the short latency discharge of neurons in PO was antagonized by 0.1% atropine (20 micrograms, icv). However, 0.1% atropine (20 micrograms, icv) did not show antagonistic effect on the inhibition of 0.1% morphine (5 micrograms, icv) at the same latency.

CONCLUSIONMT exhibited central analgesic action with mechanism different from morphine. It was suggested that the cholinergic system may be involved in analgesic process of MT.

Analgesics ; administration & dosage ; pharmacology ; Animals ; Atropine ; pharmacology ; Cats ; Electric Stimulation ; Evoked Potentials ; drug effects ; Female ; Injections, Intraventricular ; Male ; Melatonin ; administration & dosage ; pharmacology ; Morphine ; pharmacology ; Muscarinic Antagonists ; pharmacology ; Neurons ; physiology ; Splanchnic Nerves ; physiology ; Thalamic Nuclei ; drug effects ; physiology

OBJECTIVE To evaluate whether the IDO1 inhibitor 1- methyl- L- tryptophan (1- MT) combine calcium influx inhibitor carboxyamidotriazole (CAI) could further enhance the suppression of programmed death 1 (PD-1) in CD8 + T cells and investigate the curative effect of the combined use. METHODS CD8 +T cells were isolated from normal mice spleen by negative selection using magnetic cell separation. The isolated CD8 +T cells were cultured in RPMI 1640 medium containing 10% FBS and 100 U·mL- 1 IL-2 and activated by the addition of anti-CD3 and anti-CD28 (1 g·L- 1 each mabs). CD8 + T cells were pretreated for 48 h with drug and the fluo- 3 as a marker of intracellular calcium concentration was detected by flow cytometry. The calcineurin (CaN) levels were assayed with ELISA in CD8+T cells after 48 h incubation with 10 μm CAI. The nuclear translocations of NFAT and AHR were detected by immunofluorescent staining after 48 h of drug treatment. The expression of PD-1 in CD8+T cells was analyzed by flow cytometry. RESULTS Intracellular fluorescent intensity was markedly debase due to CAI treatment(P<0.01). Meanwhile, the changes of CaN content had a resembled correlation (P<0.01). Immunofluorescence experiment showed that after combination therapy the transfer of NFAT and AHR in nuclear substantially reduced. Flow cytometry revealed that after the combination caused a significant decrease in PD-1 expression in CD8+T cells. CONCLUSION CAI and 1-MT could inhibit markedly the expression of PD-1 in CD8 +T cells by inhibiting the nuclear translocation of NFAT and AHR, respectively and the combination of them has synergetic effect.

Objective To explore the correlation between anti-cyclic citrullinated peptide(A-CCP) antibody and tumor necrosis factor(TNF)-?, rheumatoid factor(RF), ESR, PLT count and clinical features in patients with rheumatoid arthritis(RA), and the outcome of unclassified arthritis(arthralgia)patients after six months follow up. The value of A-CCP antibdy in the diagnosis of early RA and its pathogenetic roles is in- vestigated. Methods A-CCP antibody and TNF-?were detected by ELISA and the RF was tested by the rate scatting immunity method in 91 RA patients, 46 unclassified arthritis(arthralgia)patients and 45 other rheumatic diseases patients. Results A-CCP antibody levels in serum correlated significantly with TNF-?levels, PLT count and the degree of joint swelling in RA and unclassified arthritis(arthralgia)patients(r= 0.854, P=0.O00; r=0.882, P=0.000; r=0.318, P=0.002; r=0.486, P=0.001; r=0.291, P=0.005; r=0.731, P= 0.000 respectively). A-CCP antibody levels in serum was weakly negatively correlated with the gripping power in RA patients(r=0.228, P=0.030). And it was weakly correlated with ESR in unclassified arthritis(arthrai- gia)patients(r=0.365, P=0.013). Compared with other rheumatic diseases patients, A-CCP antibody levlels in serum increased significantly in RA and unclassified arthritis(arthralgia)patients(P=0.000). Compared with normal controls, it increased in other rheumatic diseases patients(P=0.011). Twenty-four patients had positive A-CCP antibody in 46 unclassified arthritis(arthralgia)patients. Thirty-two out of 46 unclassified arthritis(arthralgia)patients were early RA after 6 monthes follow up. 95.8%(23/24)unclassified arthritis (arthralgia)patients with positive A-CCP antibody were early RA. Conclusion A-CCP antibody reflects disease activity in certain extent. It's benefit to the diagnosis of early RA. High A-CCPantibody levels com- bined with high levels of TNF-?, ESR, PLT count and joint swelling can help the diagnosis of early RA.

Abdominal Neoplasms ; complications ; metabolism ; pathology ; surgery ; Adult ; Dendritic Cell Sarcoma, Follicular ; complications ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Female ; Humans ; Ki-1 Antigen ; metabolism ; Leukocytosis ; complications ; metabolism ; pathology ; surgery ; Receptors, Complement 3b ; metabolism ; Receptors, Complement 3d ; metabolism ; Young Adult

OBJECTIVETo observe the effect of high glucose exposure on connective tissue growth factor (CTGF) expression in cultured human renal tubular epithelial cells and investigate the role of p38MAPK pathway in this process.

METHODSHuman renal tubular epithelial cells (HKC) with and without SB203580 pretreatment were cultured in the presence of high glucose levels for 24, 48, 72, 96 h and 20 days. RT-PCR, immunohistochemical staining, indirect fluorescence staining and Western blotting were used to detect the changes in CTGF mRNA and protein expressions in the cells after the treatment.

RESULTSLow levels of CTGF mRNA and protein were detected in cultured HKC cells, and after high glucose treatment, the mRNA expression increased gradually and reached the peak level at 48 h, then followed by gradual decrease till recovering the baseline level at 96 h. Prolonged high glucose treatment for 20 days resulted in persisted high CTGF mRNA expression twice the level in the control group. The expression level of CTGF protein also increased progressively as the treatment time then prolonged, and long-term (20 days) treatment increased the expression by 4 folds in comparison with the expression in the control cells. SB203580 significantly inhibited the increase in the expressions of CTGF mRNA and protein stimulated by high glucose treatment.

CONCLUSIONHigh glucose treatment can increase CTGF mRNA and protein expressions in cultured human renal tubular epithelial cells, suggesting that increased CTGF levels is a key event in the pathogenesis of renal tubulo-interstitial fibrosis in patients with diabetic nephropathy. p38MAPK pathway may also participate in this process.

Cells, Cultured ; Connective Tissue Growth Factor ; genetics ; metabolism ; Diabetic Nephropathies ; pathology ; Epithelial Cells ; cytology ; metabolism ; Glucose ; pharmacology ; Humans ; Kidney Tubules ; cytology ; RNA, Messenger ; genetics ; metabolism ; Signal Transduction ; p38 Mitogen-Activated Protein Kinases ; metabolism

To develop an analytic method for qualitative and quantitative analysis of dodecatetraenamides A, B in 42 samples of two official species of Asari Radix et Rhizoma( ARR) (37 samples of Asarum heterotropoides var. mandshuricum with different collection time and 5 samples of Asarum sieboldiivar. seoulense). The HPLC-IT-TOF-MS/MS methods for the qualitative and UPLC-PDA methods for the quantitative analysis were established. Dodecatetraenamides A, B were identified by comparing the retention time, UV absorption spectrum and quasi-molecular ion peak [ M + H]+ with the reference compound using HPLC-IT-TOF-MS/MS. The content of dodecatetraenamides A and B in ARR were determined by UPLC-PDA. The separation was successfully carried out on a ACQUITY UPLC BEH C18 (2.1 mm x 100 mm, 1.7 µm) column eluted with mobile phases of water (A) and acetonitrile (B) in gradient program (0-3 min, 35% B; 3-5 min, 35%-36% B; 5-6 min, 36%-43% B; 6 min-11 min 43% B; 11-12 min, 43%-100% B). The column temperature was 45 °C, and the detection wavelength was set at 254 nm. The flow rate was 0.6 mL · min(-1). On one level mass spectrometry scanning, the results showed that the quasi-molecular ion [M + H] + of both dodecatetraenamides A and B were m/z 248.20. The quantitative method with UPLC-PDA has made the baseline separation of the constituents, which were reported as mixtures in the most literatures. The average recovery of dodecatetraenamides A and B were 97.90% and 99.86%, the relative standard deviation were 0.4% and 1.1%, respectively. The contents of dodecatetraenamides A, B in all ARR samples was in the range of 0.11-3.89 and 0.24-6.65 mg · g(-1). Their contents reduced with the extension of storage time. Compared with the samples of 2013, the average content of the two constituents in the samples collected in year 2002-2003 reduced 34% and 36%, respectively (P < 0.05). Compared the A. sieboldii var. seoulense and A. heterotropoides var. mandshuricum with the same collective time and production area, the average contents of the two constituents in latter were up to (1.59 ± 0.75) mg · g(-1) and (2.90 ± 1.17) mg · g(-1), respectively, significantly higher than that in A. sieboldii var. seoulense (dodecatetraenamide A were (0.78 ± 0.52) mg · g(-1), dodecatetraenamide B were (1.69 ± 0.83) mg · g(-1)) (P < 0.05). The content of the dodecatetraenamide A in overground part was in the range of 0.11-0.33 mg · g(-1), dodecatetraenamide B was 0. 24-0.60 mg · g(-1), which were much lower than that of the underground part of ARR (dodecatetraenamide A was in the range of 0.73-3.89 mg · g(-1), dodecatetraenamide B was 2.11-6.24 mg · g(-1)). The method was certified to be simple, accurate and reliable and could be used for qualitative and quantitative analysis of dodecatetraenamide A and B in different species of ARR, also can be used for the comprehensive quality control of traditional Chinese medicine, Asari Radix et Rhizoma.
Amides ; chemistry ; Asarum ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Mass Spectrometry ; Molecular Structure ; Rhizome ; chemistry

OBJECTIVETo analyze the secular changes of physical growth in 7-17 year old students of Zang-tribe in Tibet during 1965-2004, and to make suggestions for further improvement.

METHODSThe data collected by local government in 1965 and Chinese national survey on students' physical fitness and health in 1985, 1995 and 2004 were used to analyze and find out the differences from the students' stature, body weight, chest girth, and BMI in different years, and to compare the increasing rate per 10 years in different periods of time.

RESULTSLevels on stature, body weight and chest girth were increased significantly among 7-17 year-old students of Zang-tribe in Tibet during 1965-2004. From 1965 to 1985, the levels of stature and body weight were increased 11.4 cm and 5.0 kg respectively among male Zang-tribe students, 9.1 cm and 3.9 kg among the female. During 1985-2004, stature levels of Zang-tribe students was kept rising in much less extent, while average levels of body weight were reaching up remarkably, especially in 1995-2004, in which the average increasing rate per 10 years had achieved 14.8% and 13.4% among the male and female students aged 7-17 years old respectively. Levels of chest girth were increased rapidly in 1965-1985, showing a higher increasing rate in male students (2.7%) than in female students (1.9%). In 1985-2004, the increasing rates fell obviously both in male and female students, but there existed a higher rate in 1995-2004 than in 1995-1985, especially for females (4.0%). Upon the secular changes of stature and body weight, the levels of BMI were decreased in 1985 as comparing to those in 1965, then increased rapidly in 1985-2004, especially in 1995-2004, having an increasing rate up to 12.6% and 12.4% respectively in male and female students.

CONCLUSIONSecular changes on physical growth among 7-17 year-old students of Zang-tribe in Tibet were paralleled with the economic progress in 1965-2004, giving active effects by economic progress and nutrition improvements to the physical growth of children and adolescents.

Adolescent ; Body Height ; physiology ; Body Mass Index ; Body Weight ; physiology ; Child ; Female ; Health Surveys ; Humans ; Male ; Students ; statistics & numerical data ; Tibet

Objective To learn the changes of blood lead levels and serum biochemical parameters of the school agechildren from different areas. Methods All research objects, the school age children, were from three different areasrespectively, including a mountainous area (L area), an island area (H area) where there is not history of Pb pollution,and an industry area (N area) in relation to Pb pollution. The morning urine and peripheral venous blood samples werecollected from the school age children. Pb in blood (PbB), δ-aminoaevulinic acid in urine (ALA), Ca2+, BUN, Cr inserum, and Thyroid Stimulating Hormone (TSH), thyroxin (T4), free thyroxin (FT4) levels were detected. ResultsPbB levels [M was 36.0 ppb] of the school age children from N area were significantly higher than that of L area [22.0 ppb] andH area [23.8 ppb]. On the contrary, serum Ca2+ levels of the school age children from N area were significantly lower than thatof L area and H area. Serum T4 of N area was significantly lower than that of L area and H area. Serum FT4 of H area wassignificantly higher than that of L area and N area. And TSH of N area and H area were both obviously lower than that of L area.But all of these thyroxin indexes were in the range of normal values. Conclusion It should be widely concerned that thesignificant elevation of PbB levels may have a negative impact on school age children in the future.

OBJECTIVETo investigate the autoimmune injuries of diabetic macrovascular disease (aorta) and the protective effects of immunosuppressive agent (cyclosporine A, CsA) on aortic injuries in streptozotocin (STZ)-induced diabetic rats.

METHODSSTZ-induced diabetic rats were assigned randomly to 6 groups which received low (BML or AML, 1 mgxkg(-1)xd(-1)), middle (BMM or AMM, 4 mgxkg(-1)xd(-1)) or high (BMH or AMH, 8 mgxkg(-1)xd(-1)) dose of CsA from 1 week before or after STZ for 8 weeks. Diabetic rats without any treatment, insulin-treated diabetic rats and normal rats were also monitored simultaneously and served as control groups. The pathologic abnormalities of the aorta were verified by HE, Masson staining and electronmicroscopy. The depositions of immunoglobulins (IgG, IgM and IgA) were determined by immunohistochemistry and immunofluorescence methods.

RESULTSAt the end of study, lymphocytes infiltration and collagen content (26 582 +/- 6901) were significantly higher in diabetic aorta than those in non-diabetic aorta (Collagen: 7482 +/- 3491, P < 0.01). The deposited IgG and IgA were also significantly increased in diabetic aorta compared with non-diabetic aorta (IgG: 11 789 +/- 2491 vs. 2518 +/- 1066, P < 0.01; IgA: 17 430 +/- 3159 vs. 1135 +/- 758, P < 0.01). These changes were not affected by insulin while CsA intervention significantly reduced aortic collagen content (BMH: 13 518 +/- 5440, P < 0.01 vs. STZ) and immunoglobulin deposition (BMH: IgG: 7584 +/- 4462; IgA: 6176 +/- 1900, all P < 0.01 vs. STZ). These immunoglobulin deposition changes were confirmed by results of immunofluorescence. Aortic collagen accumulation was positively correlated to aortic immunoglobulin deposition (IgG, r = 0.556, P < 0.01; IgA, r = 0.661, P < 0.01).

CONCLUSIONSOur data suggest that the autoimmune injuries might be a promoting factor in the pathogenesis of the diabetic macrovascular disease which could lead to the development of macrovascular disease. Immunosuppressive agent, such as CsA, could inhibit the abnormal deposition of immunoglobulins and therefore, delay the development of diabetic macrovascular disease in this model.

Animals ; Aorta ; immunology ; pathology ; Aortic Diseases ; etiology ; Cyclosporine ; pharmacology ; Diabetes Mellitus, Experimental ; immunology ; pathology ; Endothelium, Vascular ; drug effects ; pathology ; Immunosuppressive Agents ; pharmacology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the influence of adrenaline on the expression of TGFbeta1, bFGF and procollagen for human normal and hypertrophic scar dermal fibroblasts cultured in vitro.

METHODSHuman normal and hypertrophic scar dermal fibroblasts were propagated in a serum-free in vitro model with adrenaline for 24 hours. The human mRNA levels of bFGF, TGF-beta1 and I procollagen in fibroblasts were determined by RT-PCR. Levels of bFGF and TGF-beta1 in the supernatants of fibroblasts cultured in vitro were determined by enzyme-linked immunosorbent assay (ELISA).

RESULTSIn our study, adrenaline caused statistically significant increase in the peak levels of bFGF for normal and hypertrophic scar fibroblast cell lines (P < 0.01). It also caused statistically significant decrease in the level of TGF-beta1 for normal and hypertrophic scar fibroblast cell lines. Modulation of normal fibroblasts with 0.05, 0.10 and 0.20 micromol/L adrenaline resulted in a statistically significant (P < 0.01) decrease in the expression of I procollagen mRNA. However, only 0.20 micromol/L adrenaline can decreased the mRNA expression of I procollagen in the hypertrophic scar fibroblasts.

CONCLUSIONSWe conclude from these results that adrenaline can increase the production of bFGF and decrease production of TGF-beta1 and I procollagen in human normal dermal and hypertrophic scar fibroblasts cultured in vitro.

Cells, Cultured ; Cicatrix, Hypertrophic ; metabolism ; Collagen Type I ; metabolism ; Epinephrine ; pharmacology ; Fibroblast Growth Factor 2 ; metabolism ; Fibroblasts ; drug effects ; metabolism ; Humans ; Procollagen ; metabolism ; RNA, Messenger ; metabolism ; Transforming Growth Factor beta1 ; metabolism