Objective To study the effect of administration of transient receptor potential vanilloid 1 (TRPV1) antagonist capsazepine (CPZ) into the nucleus accumbens (NAc) on the regulation of body weight and fat accumulation in rats, and evaluate the therapeutic effect of CPZ on neuropathic obesity. Methods Forty rats were randomly divided into 5 groups (n=8), namely, control group (without giving any treatment),group A (performed intra-NAc injection of 1 nmol/mL CPZ),group B (performed intra-NAc injection of 10 nmol/mL CPZ),group C (performed injection of 10 nmol/mL CPZ via the areas adjacent to NAc) and group D (performed injection of 10 nmol/mL CPZ via the dorsal striatum); rats of the later 4 groups were given 1 μL CPZ daily for a consecutive 3 d.The body weight of rats from different treatment groups 1 and 3 weeks after the injection was measured and compared them to that of the control group.Then,the animals were sacrificed,the body fat content of each group was evaluated. Results The body weight in rats of group A 1 week after the injection increased (126.31± 10.25)％ as compared with their original weight, and that in rats of control group increased (148.78±6.98)％ as compared with their original weight, which showed significant difference between the 2 increment (P＜0.05); the body weight in rats of group B 1 week after the injection increased (115.87± 13.90)％ as compared with their original weight, which showed significant decrease as compared with that in the control group (P＜0.05); The body weight in rats of group B 3 week after the injection increased (132.82±15.8)％ as compared with their original weight, and that in rats of control group increased (164.86±6.34)％ as compared with their original weight,which showed significant difference between the 2 increment (P＜0.05).Furthermore,the body fat content in group B was significantly lower than that of other groups (P＜0.05). Conclusion Blockade of NAc TRPV1 receptor by CPZ has significant inhibitory central adjusting effect on body weight and fat content in rats; and TRPV1 antagonist may potentially serve as a specific drug for neuropathic obesity.

Field operations,work and war on plateau is an important part of future war. It is necessary to study the early treatment and evacuation of scull injury. Research and survey were made in 7 field hospitals and hundreds of records were summaried before treatment criterion in early phase was put forward for patients on plateau as well as the evacuation in different level and stage,which aims to actively save patients' lives within limited time and reduce the death rate and prevent deformity. References are provided for war,training,and construction in battlefield.

The binding function of EGF1 domain peptide with tissue factor (TF) and its ability of triggering coagulation were explored. The TF expression model in vitro was established by lipopolysaccharide induction. The affinity of EGFP-EGF1 and TF expressing cells was analyzed by fluorescence microscopy and flow cytometry (FCM). The affinity of EGFP-EGF1 and rat soluble TF was quantitated by surface plasmon resonance (SPR). The ability of EGFP-EGF1 in triggering coagulation was tested by prothrombin time assay. The FCM results showed recombinant factor VII (rFVII) could definitely depress the integration of EGFP-EGF1 with recombinant TF (rTF) (68.65%+/-3.86% vs 57.98%+/-4.71%, P<0.01). The SPR results indicated the association constant ka of EGFP-EGF1 proteins was higher than rFVII (8.29+/-1.39 vs 3.75+/-0.32, P<0.01). However, the EGFP-EGF1 protein lost the activity of triggering coagulation as compared with blood plasma of normal SD rats (56.8+/-3.2 s vs 17.8+/-3.4 s, P<0.01). It was concluded that the rat EGF1 peptide could specifically bind to TF without the ability of triggering coagulation. EGF1 peptide may be a good target head for delivering drugs to TF in anticoagulation therapy.

The efficacy and safety of recombinant tissue plasminogen activator (rtPA) need to be improved due to its low bioavailability and requirement of large dose administration.The purpose of this study was to develop a fibrin-targeted nanoparticle (NP) drug delivery system for thrombosis combination therapy.We conjugated rtPA to poly(ethylene glycol)-poly(ε-caprolactone) (PEG-PCL) nanoparticles (rtPA-NP) and investigated its physicochemical characteristics such as particle size,zeta potential,enzyme activity of conjugated rtPA and its storage stability at 4℃.The thrombolytic activity of rtPA-NP was evaluated in vitro and in vivo as well as the half-life of rtPA-NP,the properties to fibrin targeting and its influences on systemic hemostasis in vivo.The results showed that rtPA-NP equivalent to 10％ of a typical dose of rtPA could dissolve fibrin clots and were demonstrated to have a neuroprotective effect after focal cerebral ischemia as evidenced by decreased infarct volume and improved neurological deficit (P＜0.001).RtPA-NP did not influence the in vivo hemostasis or coagulation system.The half-life of conjugated rtPA was shown to be approximately 18 times longer than that of free rtPA.These experiments suggested that rtPA-conjugated PEG-PCL nanoparticles might be a promising fibrin-targeted delivery system for a combination treatment of thrombosis.

OBJECTIVETo investigate the expression of ghrelin and its receptor, growth hormone secretagogue receptor (GHS-R), in the hypothalamus and gastrointestinal tract in rats with chronic renal failure (CRF) and explore their relationship with the disorder of gastrointestinal tract motility.

METHODSSD rats were randomly divided into sham-operated group (n=8) and CRF group (n=16), and in the latter group, the rats were subjected to 5/6 nephrectomy to induce CRF. Real-time PCR and immunohistochemical staining were used to detect the distribution of mRNA and protein of ghrelin and GHS-R in the gastric fundus, duodenum, and hypothalamus.

RESULTSThe rats in the CRF group showed a significantly higher expression of ghrelin mRNA and protein in the gastric fundus but a lower expression in the hypothalamus than those in the sham-operated group (P<0.01), but the expression in the duodenum was similar between the two groups (P>0.05). The expression of GHS-R mRNA and protein in the gastric fundus was significantly higher in the CRF group than in the sham-operated group (P<0.01), while in the hypothalamus and duodenum, the expression was significantly lower in the CRF group (P<0.01).

CONCLUSIONThe different distribution patterns of ghrelin and GHS-R in the tissues may be an important pathological basis of gastrointestinal motility disorder in CRF.

Animals ; Gastrointestinal Tract ; metabolism ; Ghrelin ; genetics ; metabolism ; Hypothalamus ; metabolism ; Kidney Failure, Chronic ; metabolism ; Male ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Ghrelin ; genetics ; metabolism

OBJECTIVETo study the expression of green fluorescent protein gene and immunogenicity of ES312 vaccine both mediated by Starburst polyamidoamine (PAMAM) dendrimers in vivo.

METHODSThe complex of green fluorescent protein or ES312 gene with Starburst PAMAM dendrimers were injected intramuscularly in Balb/c mice. The expression level and distribution of green fluorescent protein gene was detected by flow cytometer, Western blot and immunofluorescence assay. The immunogenicity of DNA vaccine was detected by enzyme-linked immunosorbent assay.

RESULTSThe expression of green fluorescent protein mediated by Starburst PAMAM dendrimers was found in heart, liver, spleen, lung, kidney, brain and injected muscle from 2 hours to 7 days after the vaccination. The highest expression level of the gene was detected in kidney, as well as in endothelial cells. The antibody response evoked by the DNA vaccine carried by the Starburst PAMAM dendrimers was significantly higher than that of the net DNA vaccination. Vaccination with Starburst PAMAM dendrimers elicited higher expression level of the gene in brain and kidney than with the net gene itself.

CONCLUSIONAs a novel non-viral DNA carrier with low self-antigenicity, Starburst PAMAM dendrimers have potential to mediate DNA transfer and expression in vivo.

Animals ; Biocompatible Materials ; pharmacology ; Dendrimers ; Drug Carriers ; pharmacology ; Female ; Green Fluorescent Proteins ; genetics ; pharmacokinetics ; Malaria Vaccines ; immunology ; Mice ; Mice, Inbred BALB C ; Polyamines ; pharmacology ; Vaccination ; Vaccines, DNA ; immunology

Female ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Liver Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Tissue Array Analysis ; Tumor Suppressor Protein p53 ; metabolism

The aim of this study was to investigate if transfusion of mesenchymal stem cells (MSC) could exhibit beneficial effects on rheumatoid arthritis. Human bone marrow MSC were intraperitoneally injected into Wistar rats with collagen-induced arthritis at a dose of 10(7) on the next day (preventive group) or 2 weeks (treatment group) after collagen II induction, once a week for 2 weeks (preventive group) or 4 weeks (treatment group). The control group was given normal saline (NS) at corresponding time. The symptom scorings were documented weekly from the second week of the induction. On week 6, the hind joints of the rats were pathologically examined and the activation status of splenocytes was analyzed by flow cytometry. The results showed that all the rats developed arthritis and subsequent joint abnormality. On the sixth week, symptom scores of the rats that received MSC preventive (9.5 ± 0.5) or therapeutic (9.4 ± 0.6) infusions had no significant difference between each other, but were significantly greater than those of the NS controls (7.6 ± 0.6, P < 0.05). Consistently, pathological examination on the involved knees showed that the synovitis and arthritis scorings of MSC treated rats were greatly elevated compared with NS controls. Furthermore, the ratios of CD86(+) cells in the spleens of MSC prevention, MSC treatment and NS control groups were (4.16 ± 1.48), (4.06 ± 1.97) and (4.15 ± 2.04) respectively, while those of CD11b/c(+)CD86(+) cells were (1.04 ± 0.68), (0.95 ± 0.56) and (0.98 ± 0.44), all of which were significantly higher than those of healthy controls [(0.97 ± 0.18) and (0.30 ± 0.17), P < 0.05 for both parameters]. It is concluded that MSC infusion has little beneficial effects on collagen-induced arthritis in rats, conversely, MSC therapy aggravated the damage of the involved joints, its underlying mechanisms need to be further investigated.
Animals ; Arthritis, Experimental ; pathology ; prevention & control ; therapy ; Bone Marrow Cells ; Cells, Cultured ; Humans ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; cytology ; Rats ; Rats, Wistar ; Transplantation, Heterologous ; Treatment Outcome

BACKGROUNDCombination paclitaxel and carboplatin is currently a first-line regimen for ovarian cancer. However, many patients develop tumor recurrence or drug resistance to this regimen. The study aims to investigate the effectiveness and safety of an oxaliplatin + epirubicin + ifosfamide regimen for the treatment of recurrent and drug-resistant epithelial ovarian cancer.

METHODSA retrospective analysis of 73 patients with recurrent and drug-resistant ovarian cancer was performed; 38 cases of them received oxaliplatin + epirubicin + ifosfamide regimens (IAP group), 35 patients received non-oxaliplatinbased chemotherapy regimens (control group). The therapeutic effects and side effects of the oxaliplatin + epirubicin + ifosfamide regimen were analyzed and summarized. Kaplan-Meier survival curves and Cox proportional hazards regression were used to compare progression-free and overall survival between the two groups.

RESULTSOf the 38 patients in the IAP group, 14 patients (36.84%) achieved complete remission, 12 (31.58%) achieved partial remission, 2 (5.26%) achieved stable disease and 10 (26.32%) developed progressive disease. The overall effective rate (complete or partial remission) of the IAP regime was 68.42%. While, of the 35 patients in the control group, 12 patients (34.29%) achieved complete remission, 3 (8.57%) achieved partial remission, 5 (14.29%) achieved stable disease and 15 (42.86%) developed progressive disease. The overall effective rate was 42.86%, which was lower than that in the IAP group (P = 0.035, χ(2) = 4.836). Progression-free survival was 9.5 months (0-64 months) in the IAP group vs. 3 months (0-74 months) in the non-oxaliplatin group (P = 0.014 by Kaplan-Meier survival curves; HR = 2.260; 95%CI 1.117-4.573; P = 0.023 by Cox proportional hazards regression). Median overall survival was 46 months (9-124 months) in the IAP group vs. 35 months (9-108 months) in non-oxaliplatin group (P = 0.018 by Kaplan-Meier survival curves; HR = 2.272; 95%CI 1.123-4.598; P = 0.022 by Cox proportional hazards regression). In IAP group, 15.79% (6/38) of the patients suffered grade III-IV bone marrow arrest. The main non-hematological side effects of the IAP regimen included nausea and vomiting (21.05%, 8/38), peripheral neurotoxicity (15.79%, 6/38) and hepatic or renal lesions (2.63%, 1/38). The main side effects of the two chemotherapy regimens showed no statistical difference.

CONCLUSIONThe oxaliplatin-based IAP regimen is potentially effective for salvage chemotherapy in patients with recurrent and drug-resistant ovarian cancer, with a better therapeutic effect and tolerable side effects.

Adult ; Aged ; Antineoplastic Agents ; therapeutic use ; Drug Resistance, Neoplasm ; Drug Therapy, Combination ; Female ; Humans ; Middle Aged ; Neoplasms, Glandular and Epithelial ; drug therapy ; Organoplatinum Compounds ; therapeutic use ; Ovarian Neoplasms ; drug therapy ; Platinum ; therapeutic use ; Retrospective Studies

In past decade, hydrogen sulfide (H₂S) as a novel gasotransmitter, covered many fields in biological and medical research. However, there is no effective, convenient and high-throughput method for determination of circulatory H₂S until now. Here, we aim to develop an easy method for measurement of circulatory H₂S by modified methylene blue method. In the present study, we added Zn²⁺ to plasma sample to deposit H₂S, HS⁻ and S²⁻, as well as plasma protein, then used NaOH to re-dissolve plasma protein. ZnS deposition was re-dissolved by the addition of N, N-dimethyl-p-phenylenediamine, and the remnant protein was deposited by trichloroacetic acid. After centrifugation, ferriammonium sulfate was added to the supernatant fluid to generate methylene blue, which was analyzed by spectrophotometer at 665 nm. Using the present method, we found that most ions including sulfate and thiosulfate did not affect detection of H₂S concentration, but albumin (physiological concentration) reduced the detection value, which suggested the binding of serum albumin and a certain amount of H₂S. The relative recovery ratio of present method is 81.9%, which implies that the method is relative accurate for the determination of H₂S concentration in plasma or serum. H₂S levels of frozen plasma samples from 65 healthy volunteers detected by the present method were (13.93 ± 4.98) µmol/L. There was no obvious difference between the detection values of fresh and frozen samples from the same SD rats. These results suggest the modified methylene blue assay is stable, sensitive, convenient and high-throughput. The method can be used to analyze the circulatory H₂S in clinical and basic research.
Animals ; Blood Chemical Analysis ; methods ; Humans ; Hydrogen Sulfide ; blood ; Methylene Blue ; chemistry ; Phenylenediamines ; chemistry ; Rats ; Rats, Sprague-Dawley ; Sulfides ; chemistry ; Zinc Compounds ; chemistry