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ObjectiveTo investigate the application value of Qingre Huashi Sanjie enema prescription in the treatment of the patients with sequelae of pelvic inflammatory disease (syndrome of combined dampness,heat,and stasis) and the effects of this prescription on inflammatory mediators and T lymphocyte subsets. MethodThe patients with sequelae of pelvic inflammatory disease (syndrome of combined dampness,heat,and stasis) treated from May 2022 to August 2023 were included in this study and randomized into two groups (79 cases). The control group was treated with conventional Western medicine,and the observation group was treated with Qingre Huashi Sanjie enema prescription on the basis of the therapy in the control group. Both groups were treated for 12 weeks. The serum levels of monocyte chemoattractant protein-1 (MCP-1),transforming growth factor-β1 (TGF-β1),and interleukin-6 (IL-6) were measured by enzyme linked immunoserbent assay (ELISA) before and after treatment in both groups. The erythrocyte sedimentation rate (ESR) and fibrinogen (FIB) were measured by an automatic blood rheology analyzer before and after treatment in both groups. The serum levels of CD4+,CD4+/CD8+ before and after treatment in both groups were measured by flow cytometry. The traditional Chinese medicine (TCM) symptom score and the 36-item short form survey (SF-36) score were assessed before and after treatment. The uterine artery resistance index (RI),uterine artery pulsatility index (PI),and uterine artery peak systolic velocity (PSV) were measured by Doppler before and after treatment. The clinical efficacy and the occurrence of adverse reactions were compared between the two groups. ResultAfter treatment,the levels of MCP-1,TGF-β1,IL-6,ESR,and FIB decreased in both groups (P<0.01),and the decreases were larger in the observation group than in the control group (P<0.05,P<0.01). After treatment,the serum levels of CD4+ and CD4+/CD8+ elevated in both groups (P<0.01) and the observation group had higher levels of CD4+ and CD4+/CD8+ than the control group (P<0.05,P<0.01). The treatment in both groups decreased the TCM symptom score and TCM sign score and increased the SF-36 score (P<0.01),and the changes were more significant in the observation group than in the control group (P<0.05,P<0.01). In addition,the treatment lowered RI and PI and elevated PSV (P<0.01),and the changes in these indicators were more significant in the observation group than in the control group (P<0.01). The total response rate in the observation group was 93.67% (74/79),which was higher than that (79.75%,63/79) in the control group (χ2=6.645,P<0.05). There was no significant difference in the occurrence of adverse reactions between the two groups. ConclusionFor the patients with sequelae of pelvic inflammatory disease (syndrome of combined dampness,heat,and stasis),Qingre Huashi Sanjie enema prescription can reduce inflammation,attenuate hypercoagulability,improve hemodynamics,and regulate the immune function,demonstrating a definite therapeutic effect.
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Mounting evidence supports an important role of chemokines, produced by spinal cord astrocytes, in promoting central sensitization and chronic pain. In particular, CCL2 (C-C motif chemokine ligand 2) has been shown to enhance N-methyl-D-aspartate (NMDA)-induced currents in spinal outer lamina II (IIo) neurons. However, the exact molecular, synaptic, and cellular mechanisms by which CCL2 modulates central sensitization are still unclear. We found that spinal injection of the CCR2 antagonist RS504393 attenuated CCL2- and inflammation-induced hyperalgesia. Single-cell RT-PCR revealed CCR2 expression in excitatory vesicular glutamate transporter subtype 2-positive (VGLUT2) neurons. CCL2 increased NMDA-induced currents in CCR2/VGLUT2 neurons in lamina IIo; it also enhanced the synaptic NMDA currents evoked by dorsal root stimulation; and furthermore, it increased the total and synaptic NMDA currents in somatostatin-expressing excitatory neurons. Finally, intrathecal RS504393 reversed the long-term potentiation evoked in the spinal cord by C-fiber stimulation. Our findings suggest that CCL2 directly modulates synaptic plasticity in CCR2-expressing excitatory neurons in spinal lamina IIo, and this underlies the generation of central sensitization in pathological pain.
Subject(s)
Animals , Female , Male , Mice , Benzoxazines , Pharmacology , Therapeutic Uses , Chemokine CCL2 , Genetics , Metabolism , Pharmacology , Excitatory Amino Acid Agents , Pharmacology , Excitatory Amino Acid Agonists , Pharmacology , Freund's Adjuvant , Toxicity , Hyperalgesia , Metabolism , Long-Term Potentiation , Physiology , Luminescent Proteins , Genetics , Metabolism , Mice, Inbred C57BL , Mice, Transgenic , Myelitis , Drug Therapy , Metabolism , Neurons , Pain Management , Somatostatin , Genetics , Metabolism , Spinal Cord , Cell Biology , Spiro Compounds , Pharmacology , Therapeutic Uses , Vesicular Glutamate Transport Protein 2 , Genetics , Metabolism , Vesicular Inhibitory Amino Acid Transport Proteins , Genetics , MetabolismABSTRACT
Objective To investigate the relation of Toll-like receptor 4 (TLR4)and monocyte chemoattractant protein-1 (MCP-1)with acute coronary syndrome.Methods 60 patients with coronary heart disease patients including 39 cases of a-cute coronary syndrome (ACS group),21 cases of stable angina (SA group)from January 2015-March 2016 in Xi’an First Hospital were in this observation.ACS group was divided into acute myocardial infarction (AMI,n=21)and unstable angina pectoris (UAP,n=18).Another 30 cases with angiographically normal was included in control group.Enzyme-linked immu-nosorbent assay (ELISA)detection was used to detect the serum content of MCP-1,TLR4 expression was tested by flow cy-tometry.Triglyceride (TG),total cholesterol (TC),low-density lipoprotein cholesterol (LDL-C)and high-density lipopro-tein cholesterol (HDL-C)were detected.Results TLR4 levels of ACS group was significantly higher than SA group (t=4.455,P=0.021).Serum MCP-1 levels of ACS group were significantly higher than SA group (t=3.220,P=0.002)and control group (t=6.197,P=0.000).MCP-1 levels of ACS group were higher than control group (t=2.306,P=0.025). The Spearman correlation test TLR4 was positively with MCP-1 (r=2.389,P=0.025).Conclusion MCP-1 and TLR4 may determine the severity of coronary heart disease,combined detection of MCP-1 and TRL4 can provide the clinical value for early diagnosis and treatment of ACS.
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Objective To investigate the effect of on serum and urine MCP -1 secretion in patients with Stage IV Type 2 diabetic nephropathy treated by lipoic acid .Methods We enrolled 76 diabetic nephropathy patients ,who were randomly divided into two groups . Patients in group T (24 males and 14 females) were treated by lipoic acid 0.6 gram per day.Those in group C (22 males and 16 fe-males) were treated by routine drugs.The serum creatinine (Scr), urea nitrogen (BUN), fasting blood sugar(FBS) and 24h urinary pro-tein,serum and urine MCP -1 secretion were measured at the experiment onset and 3 weeks later.Results There was no significant difference in the serum creatinine , urea nitrogen, fasting blood sugar, HbA1c between the two groups neither at the experiment onset nor after 3 weeks.Compared to experiment onset , 24h urine protein (Tp/24h), serum and urine MCP-1 secretion were all significantly de-creased (P<0.05) in group T after 3 weeks.Compared to group C (2.41 ±0.91g/24h, 91.45 ±33.41pg/ml, 114.78 ±36.35pg/ml), all the levers in group T (1.89 ±0.72g/24h, 39.50 ±13.68pg/ml, 63.41 ±19.57pg/ml) was significantly decreased (P<0.05) after 3 weeks.There was a positive correlation between the serum , urine MCP-1 levels and Tp/24h (r=0.572, P<0.05;r=0.697,P<0.05).Conclusion Lipoic acid can reduce urine protein excretion in diabetic nephropathy patients , maybe by decreasing serum and u-rine MCP-1 secretion .
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Objective To investigate the changes of monocyte chemoattractant protein-1(MCP-1)in serum and urine of lupus nephritis(LN)patients in active phase and remission phase.Methods The levels of MCP-1 in serum and urine of 58 LN patients(27 of active phase and 31 of remission phase)were measured by ELISA.The correlation between the levels of MCP-1 in variant phase of LN and other relevant factors were analyzed.Results The MCP-1 levels in sera of both active phase and remission phase of LN patients were markedly higher than those in controls(548.5?347.2 ng/L and 469.1?298.4 ng/L vs 273.3?146.7 ng/L,P0.05).Conclusion The MCP-1 levels in urine of LN patients is more suitable to evaluate the activity of disease as a sensitive marker.
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Objective Rapamycin (RAPA) is an anti-proliferative immunosuppressant and has been used to suppress rejection of transplanted organs. In present study, we observed the effect of rapamycin on epithelial-myofibroblast transition (EMT)of cultured HKC cells in vitro. Methods Cultured human proximal tubular epithelial cells (HKCs) were divided into three groups: blank control, treated with TGF-?1 (1 ?g/L) and treated with TGF-?1 (1 ?g/L) plus rapamycin (0.1, 1, 10, 100 ?g/L). The protein and mRNA for ?-SMA and E-cadherin in HKC cells were determined by Western Blot and RT-PCR.The mRNA level of Snail in HKC was detected by RT-PCR. Results Rapamycin dramatically abrogated TGF-?1 induced ?-SMA expression and restored E-cadherin expressionin HKC cells in a dose-dependent manner. At a concentration of 100 ?g/L, rapamycin almost completely blocked ?-SMA mRNA and protein expression induced by TGF-?1(1 ?g/L). Rapamycin also suppressed expression of ?-SMA in HKC cells at both mRNA and protein level in a time dependent manner.We also found rapamycin dramatically abrogated TGF-?1 induced Snail mRNA expression in HKC cells in a dose-dependent manner. Conclusion Rapamycin may inhibit EMT of tubular cells in vitro. The downregulation of Snail expression might be one of the mechanisms of rapamycin blocking EMT.
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BACKGROUND: Monocyte chemoattractant protein-1 (MCP-1) belongs to C-C subfamily of chemokines, which stimulates the migration of monocytes. MCP-1 exerts various effects on the monocytes, including the induction of integrin and tissue factor, and synthesis of proinflammatory cytokines and arachidonic acid. In this study, we measured the MCP-1 levels in patients with Behcet's disease and evaluated the associations between the levels of MCP-1 and the level of other chemokines and various clinical features of Behcet's disease. METHODS: Serum samples were obtained from 67 patients with Behcet's disease and 30 healthy controls. Simultaneously, whole blood was isolated from patients (n=25) with Behcet's disease and healthy controls (n=11) and cultured in 24 well plates for 48 hours in the absence or presence of lipopolysaccharide (LPS) 5 microgram/mL, phytohaemagglutinin (PHA) 5 microgram/mL, phorbol 12-myristate 13-acetate (PMA) 50 ng/mL + ionomycin 5 microgram/mL. The MCP-1 concentrations were measured in the sera and culture supernatants by enzyme-linked immunosorbent assay (ELISA). RESULTS: The levels of serum MCP-1 were 2.5 times higher in patients with Behcet's disease than healthy controls. The patients with Behcet's disease had also higher levels of MCP-1 in the culture supernatants of whole blood cells, stimulated with LPS, but not with either PHA or PMA plus ionomycin, compared to healthy controls. Serum MCP-1 levels (n=67) were strongly correlated with serum RANTES, MIP-1alpha, IL-8 levels in Behcet's disease. In addition, the production of MCP-1 by whole blood culture from Behcet's disease patients (n=25) were also correlated well with those of RANTES, MIP-1alpha, and IL-8, when stimulated with LPS. However, MCP-1 levels in the sera and culture supernatants did not show any association with various clinical features of Behcet's disease including oral ulcer, genital ulcer, erythema nodosum, arthritis, uveitis, intestinal involvement, central nervous system involvement, and vascular thrombosis. CONCLUSION: In the sera and culture supernatants of whole blood stimulated with LPS, MCP-1 levels were higher in patients with Behcet's disease than controls and correlated well with RANTES, MIP-1alpha, IL-8 levels. These results suggest that the activation and migration of monocytes triggered by the increased production of MCP-1 may play a role in the pathogenesis of Behcet's disease.
Subject(s)
Humans , Arachidonic Acid , Arthritis , Blood Cells , Central Nervous System , Chemokine CCL2 , Chemokine CCL3 , Chemokine CCL5 , Chemokines , Cytokines , Enzyme-Linked Immunosorbent Assay , Erythema Nodosum , Interleukin-8 , Ionomycin , Monocytes , Oral Ulcer , Thromboplastin , Thrombosis , Ulcer , UveitisABSTRACT
Objective: To optimize the expression of recombinant human monocyte chemoattractant protein (rhuMCP-1) in E.coli DE3. Methods:With NBS-MICROS 15 L T.DR fermentor, pGEX-IN/huMCP-1 was constructed by our laboratory. Four parameters including pH,temperature,agitation rate and concentration of IPTG were studied by orthogonal experimental design. Results: It was found that the expression level was greatly affected by the amount of dissolved oxygen. This indicated that the agitation rate and ventilation amount were the most important parameters during fermentation. Examined by SDS-PAGE and gel scanning, the expression level of total protein was over 40% when agitation rate was 300 r/min and ventilation amount was 10 L/min. Conclusion: A method for high-level expression of huMCP-1 on pilot-scale is established, and it will be useful for large-scale industrial production of target protein.
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Objective To explore the molecular basis of vascular endothelial cell's injury by cellular S-adenosylhomocysteine (SAH) accumulation, and the mechanism for cardiovascular disease induced by dietary high methionine intake. Method After human umbilical vein endothelial cells (HUVEC) were treated without (normal) or with different concentrations of potent S-adenosylhomocysteine hydrolase (SAHH) inhibitor 3-deazaadenosine (DZA) for 24, 48 or 72h, the level of intracellular inflammatory monocyte chemoattractant protein-1 (MCP-1) was measured with enzyme linked immunosorbent assay (ELISA). Then, the relative expression of MCP-1 mRNA was detected with quantitative reverse transcription polymerase chain reaction (qRT-PCR). Furtheremore, the methylated state of MCP-1 gene promoter was analyzed with methylation special PCR (MSP) method. Results The level of intracellular MCP-1 was increased in HUVEC incubated with DZA than normal (P