Objective To investigate the relationship between the density of langerhan cell(Lc) and Tcell (Tc) and the histological classification, biological behavior and prognosis of esophageal carcinoma.Methods The immunohistochemical (ABC) method with S-100 and CD3 staining was used in samples ofprimary esophageal cancer in 249 cases for quantitative analysis. All cases were followed up for more than7 years. We observed the morphology,number and distribution of langerhan cell (Lc) and T cell (Tc) indifferent portionof the tumor mass. Results Lc positive expressed in the nucleus and cytoplasma. To positiveexpressed in cytomembrane. The number of Lc in cancer nest had positive correlation with malignantdegree of tumor, infiltration depth and metastasis of lymph node, and had negative correlation with thesurvival period. On the contrary, the number of To and Lc in peritumor interstitial tissue had negativecorrelation with the survival period. Conclusion The distribution density of Lc and Tc in cancer nest andperitumor interstitial tissue might be used as an immunological parameter for assessment of the prognosis.

Objective To detect and analyze the expression of ER and PR in endometrial benign and malignant tumor,and to study its correlation of ER and PR with the establishment and development of endometrial carcinoma.Methods The expressions of ER and PR were examined by immunohistochemical method in 58 eases of endometrial carcinoma,37 cases of atypism endometrial hyperplasia,25 cases of simple endometrial carcinoma.and 25 eases of normal endometrium.Results The expressions of ER and PR were higher in normal endometrium,atypism endometrial hyperplasia and endometrial carcinoma.They were lower in endometrial carcinoma than in atypism endometrial hyperplasia,both were higher than that in normal endometrium(P＜0.05).The rates of expression in ER and PR increased gradually from histological grade Ⅰ,Ⅱto Ⅲ.The expressions of ER and PR in histological grade Ⅰ were significantly different from that in histologic grade Ⅲ(P＜0.05).There were no correlation between expressions and ages,which expression rates less than 50 years were higher than those above 50 years.Its expression W88 not related to different clinic grade(P＞0.10).Conclusion The expressions of ER and PR increased gradually from normal endometrium,atypism endometrial hyperplasia to endometrial carcinoma.ER and PR expressions were obviously related to histologic degree.It maybe related to establishment and development of endometrial carcinoma.

OBJECTIVETo investigate the intervention effect and mechanism of compound Ginkgo biloba (CGB) preparations on nonalcoholic fatty liver disease (NAFLD).

METHODThe C57BL/6 mouse NAFLD model was induced with high fat diets. Since the 2nd week after modeling, the mice were orally administered with 600 and 200 mg x kg(-1) x d(-1) CGB for eight weeks. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG), cholesterol (CHOL) and LPS in serum, as well as pathological changes and expression of tumor necrosis factor-alpha (TNF-alpha) in hepatic tissues were observed. Changes in intestinal tight junction proteins ZO-1, Occludin, Claudin-1 in intestinal tissues were determined under microscopy.

RESULTCompared with the normal group, the model group showed obvious fatty degeneration in rat livers, with notable increase in TNF-alpha expression (P < 0.01), significant increases in ALT, AST, TG, CHOL and LPS in serum (P < 0.01, P < 0.05), injury in intestinal tight junction proteins, and remarkable declines in ZO-1, Occludin and Claudin-1 (P < 0.01). Compared with the model group, CGB high and low dose groups showed obvious relieves in fatty degeneration in rat livers and injury in intestinal tight junction proteins, significant reductions in TNF-alpha expression (P < 0.01, P < 0.05) and AST, TG, CHOL and LPS in serum (P < 0.01, P < 0.05) and remarkable increases in ZO-1 and Occludin expressions (P < 0.05).

CONCLUSIONCGB can protect intestinal tight junction proteins, reduce intestinal leakage, relieve fatty degeneration and inflammations in livers and prevent NAFLD occurrence and development.

Alanine Transaminase ; genetics ; metabolism ; Animals ; Aspartate Aminotransferases ; genetics ; metabolism ; Cholesterol ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Fatty Liver ; drug therapy ; enzymology ; genetics ; metabolism ; Ginkgo biloba ; chemistry ; Humans ; Male ; Mice ; Mice, Inbred C57BL ; Triglycerides ; metabolism

Objective To observe the effects of electrical acupuncture for the treatment of neurogenic detrusor overactivity (DO) caused by spinal cord injury (SCI).Methods Twenty SCI patients with DO were included in this study. There were 18 males and 2 females;age ranged from 17 to 58 years. Patients were given electro-acupuncture treatment at the bilateral S3 foramen at the lateral position during the vidio-urodynamic investigation when the detrusor of the patients began to contract.The different intensities of electrical stimulation were used when the DO appeared during cystometry and the most effective intensity to inhibit DO was determined. Then, the bladder was emptied and the stimulation with selected intensity was used at the beginning of cystometry. The changes of parameters in the urodynamics, the urinary incontinence times per day and the pad used per day were recorded. The acute effects were observed. Ten days were set as one course and after 3 courses and 9 courses the patients underwent urodynamic test again. The long-term effects were observed.Results After treatment, the urinary incontinence times per day and the pad used per day decreased obviously (P<0. 05). The bladder capacity increased significantly and the maximum intravesical pressure decreased significantly. There were 3 patients having the long terms treatment. Conclusions The Chinese electro-acupuncture at S3 foramen in the SCI patients with DO is demonstrated effective. After the treatment bladder capacity could increase and the times of the urinary incontinence per week decrease.

Objective To observe the effect of Zusanli acupoint injection of Astragalus Membranaceus Injection on patients with tumor leukopenia after radiotherapy and chemotherapy and its influence on cellular immune function.Methods According to clinical case of random table,60 cases of malignant tumor patients in Hubei province Xiangyang city hospital from March 2010 to August 2011 were randomly divided into two groups of 30 patients in each group.The treatment group was treated with Zusanli acupoint injection of Astragalus Injection therapy,while the control group was taken Leucogen,vitaminB4,batyl alcohol orally.Venous blood and peripheral white blood cell were taken in both groups before and after treatment.Direct immunofluorescence staining and flow cytometry were used for the detection of peripheral blood T lymphocyte total number and its subsets.Results ① clinical efficacy:Total effectiveness was 77.5％in the treatment group,and 52.5％ the control group,the difference was statistically significant between the two groups (x2=8.027,P＜0.05).Comparison of the white blood cell count and T lymphocyte subsets of peripheral blood:the white blood cell count was (3.56～4.21) × 109/L in the treatment group,and (2.89～3.39) × 109/in the control group,there was significant difference between two groups (t=2.435,P＜0.05); there was significant elevation of CD3 +,CD4+,CD4+/CD8 + [(62± 1.63) ％,(43 ± 0.80) ％,(1.48 ± 0.23) in the treatment group after the treatment comparing with (48± 1.25)％,(31 ± 0.74)％,(0.93± 0.12) before the treatment (P＜0.05 or 0.01),while there was no such changes in the control group.Conclusion Zusanli acupoint injection of Astragalus Injection may modulate the cellular immune function of patients with cancer.It is effective in the treatment of leukopenia after radiotherapy and chemotherapy.

BACKGROUND: Paraquat (PQ) is an effective herbicide and is widely used in agricultural production, but PQ poisoning is frequently seen in humans with the lung as the target organ. Clinically pulmonary pathological changes are often used to predict the severity and prognosis of the patients. In this study, we observed the expression of heat shock protein 70 (HSP70) in rat lung after PQ poisoning and to investigate the therapeutic effects of ulinastatin. METHODS: Seventy-two adult healthy SD rats were randomly divided into a control group (group A, n=24), a poisoning group (group B, n=24), and an ulinastatin group (group C, n=24). The rat models of acute PQ poisoning were established by intra-gastric administration of 80 mg/kg PQ to rats of groups B and C, and the rats of group C were intra-peritoneally injected with 100000 IU/kg ulinastatin 30 minutes after poisoning. The expression of HSP70 in lung tissue was observed, and W/D and histopathological changes in the lung tissue were compared 12, 24, 48 and 72 hours after poisoning. The expression of HSP70 in the lung tissue was assayed by using RT-PCR. All quantitative data were processed with one-way analysis of variance to compare multiple sample means. RESULTS: Compared to group A, the expression of HSP70 in the lung of rats in groups B and C increased significantly at all intervals (P<0.05). The pathological changes in lung tissue of rats with PQ poisoning included congestion, leukocytes infiltration and local hemorrhage, whereas those of group C were significantly lessened. CONCLUSION: Ulinastatin may ameliorate acute lung injury to some extent after PQ poisoning in rats by enhancing the expression of HSP70.

BACKGROUND: Paraquat (PQ) is an effective herbicide and is widely used in agricultural production, but PQ poisoning is frequently seen in humans with the lung as the target organ. Currently, there are many studies on lung injury after PQ poisoning. But the kidney as the main excretory organ after PQ poisoning is rarely studied and the mechanisms of this poisoning is not very clear. In this study, we observed the expression of caspase-3 and livin protein in rat renal tissue after PQ poisoning as well as the therapeutic effects of ulinastatin. METHODS: Fifty-four Sprague-Dawley (SD) rats were randomly divided into three experimental groups: control group (group A), paraquat poisoning group (group B) and ulinastatin group (group C), with 18 rats in each group. Rats in group B and group C were administered intragastrically with 80 mg/kg PQ, rats in group C were injected peritoneally with 100000 U/kg ulinastatin once a day, while rats in group A were administered intragastrically with the same volume of saline as PQ. At 24, 48, 72 hours after poisoning, the expression of livin in renal tissue was detected by Westen blotting, the expression of caspase-3 was detected by immunohistochemistry, and the rate of renal cell apoptosis was tested by TUNEL detection. The histopathological changes were observed at the same time. RESULTS: Compared to group A, the expression of caspase-3 in the renal tissue of rats in groups B and C increased significantly at any time point. Compared with group B, the expression of caspase-3 in renal tissue of rats in group C decreased. Compared with group A, the expression of livin in renal tissue in rats of groups B and C increased significantly at any time point (P<0.01), especially in group C (P<0.01). TUNEL method showed that the rate of renal cell apoptosis index was higher in group B at corresponding time points than in group A (P<0.01), and was lower in group C at corresponding time points than in group B (P<0.01). CONCLUSION: UTI has a protective effect on the renal tissue of rats after paraquat poisoning through up-regulating the expression of livin and down-regulating the expression of caspase-3, but the regulation path still needs a further research.

OBJECTIVETo study the effect of Mudan Granule (MD) on the glucose metabolism and beta cell function in monosodium glutamate (MSG) induced obese mice with insulin resistance (IR).

METHODSMSG obese mice were induced by subcutaneous injecting MSG (4 g/kg for 7 successive days in neonatal ICR mice). Forty MSG mice with IR features were recruited and divided into four groups according to body weight, fasting blood glucose, triglyceride (TG), total cholesterol (TC), and the percentage of blood glucose decreased within 40 min in the IR test, i.e., the model group (Con), the low dose MD group, the high dose MD group, and the Metformin group (Met). Besides, another 10 ICR mice were recruited as the normal control group (Nor). The water solvent of 2.5 g/kg MD or 5 g/kg MD was respectively administered to mice in the low dose MD group and the high dose MD group. Metformin hydrochloride was given to mice in the Met group at 0.2 g/kg body weight. Equal dose solvent distilled water was administered to mice in the Nor group and the Con group by gastrogavage, once per day. All medication was lasted for 15 weeks. Insulin tolerance test (ITT) and oral glucose tolerance test (OGTT) were performed after 6 weeks of treatment. Beta cell function was assessed by hyperglycemic clamp technique. The morphological changes in the pancreas were evaluated by hematoxylin-eosin (HE) staining. Changes of iNOS, NF-kappaB p65, and p-NF-kappaB p65 in the pancreas were tested.

RESULTSCompared with the Nor group, the blood glucose level, AUC, and fasting blood insulin, ONOO-contents, iNOS activities, and the expression of iNOS, NF-kappaB p65 subunit, pNF-kappaB p65 subunit obviously increased; decreased percentage of blood glucose within 40 min in ITT, glucose infusion rate (GIR), Clamp 1 min insulin, and Max-Insulin obviously decreased in the Con group (P < 0.05, P < 0.01). Compared with the Con group, the aforesaid indices could be improved in the Met group (P < 0.05, P < 0.01). In the low dose MD group, AUC, iNOS activities, and the expression of iNOS and p-NF-kappaB p65 subunit obviously decreased; percentage of blood glucose within 40 min in ITT and GIR obviously increased (P < 0.05, P < 0.01). In the high dose MD group, AUC, ONOO-contents, iNOS activities, and the expression of iNOS, NF-kappaB p65 subunit, and p-NF-KB p65 subunit obviously decreased; percentage of blood glucose within 40 min in ITT, Max-Insulin, and GIR obviously increased (P < 0.05, P < 0.01).

CONCLUSIONMD could significantly improve IR and functional disorder of 3 cells in MSG obese mice, which might be associated with lowering inflammatory reaction in the pancreas.

Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Female ; Insulin Resistance ; Insulin-Secreting Cells ; drug effects ; metabolism ; Male ; Metformin ; pharmacology ; Mice ; Mice, Inbred ICR ; Mice, Obese ; Obesity ; chemically induced ; metabolism ; Pancreas ; cytology ; drug effects ; Sodium Glutamate

This study is to evaluate the effects of the metformin (Met) on β cell function of diabetic KKAy mice. Female diabetic KKAy mice selected by insulin tolerance test (ITT) were divided randomly into two groups. Con group was orally administered by gavage with water, Met group with metformin hydrochloride at a dose of 0.2 g x kg(-1) for about 12 weeks. ITT and glucose tolerance tests (OGTT) were determined. Beta cell function was assessed by hyperglycemic clamp. Pancreatic biochemical indicators were tested. The changes of gene and protein expression in the pancreas and islets were also analyzed by Real-Time-PCR and immunostaining. Met significantly improved glucose intolerance and insulin resistance in KKAy mice. Fasting plasma glucose and insulin levels were also decreased. In addition, Met markedly increased glucose infusion rate (GIR) and elevated the Ist phase and maximum insulin secretion during clamp. It showed that Met decreased TG content and iNOS activities and increased Ca(2+) -Mg(2+)-ATPase activity in pancreas. Islets periphery was improved, and down-regulation of glucagon and up-regulated insulin protein expressions were found after Met treatment. Pancreatic mRNA expressions of inflammation factors including TLR4, NF-κB, JNK, IL-6 and TNF-α were down-regulated, p-NF-κB p65 protein levels also down-regulated by Met. And mRNA expressions of ion homeostasis involved in insulin secretion including SERCA2 and Kir6.2 were up-regulated by Met. Met increased SIRT5 expression level in pancreas of KKAy mice under the hyperglycemic clamp. These results indicated that chronic administration of Met regulated pancreatic inflammation generation, ion and hormone homeostasis and improved β cell function of diabetic KKAy mice.
Animals ; Blood Glucose ; Diabetes Mellitus, Experimental ; drug therapy ; Down-Regulation ; Female ; Glucose Tolerance Test ; Homeostasis ; Inflammation ; drug therapy ; Insulin ; secretion ; Insulin Resistance ; Insulin-Secreting Cells ; drug effects ; Interleukin-6 ; metabolism ; Metformin ; pharmacology ; Mice ; NF-kappa B ; metabolism ; Pancreas ; drug effects ; Tumor Necrosis Factor-alpha ; metabolism

BACKGROUND:Exosomes are membrane vesicles secreted by mesenchymal stem cells. Increasing studies have shown that mesenchymal stem cells can secrete exosomes via paracrine function to play a role in tissue injury. However, reports on how to isolate and identify exosomes derived from human umbilical cord blood mesenchymal stem cells are few. OBJECTIVE:To extract, purify and identify exosomes derived from human umbilical cord blood mesenchymal stem cells. METHODS:The cellculture supernatant of human umbilical cord blood mesenchymal stem cells was col ected. Exosome was extracted and purified with ultrafiltration and gradient centrifugation methods. The morphology of exosome was observed by transmission electronic microscope, and the expressions of CD63, CD81, CD90, CD73, CD105, CD29, and CD166 in exosome of mesenchymal stem cells were analyzed by fluorescent activated cellsorting. RESULTS AND CONCLUSION:Mesenchymal stem cells from human umbilical cord blood secreted exosome which exhibited el iptic or saucer-like shape and its diameter ranged from 40 to 100 nm with membrane structure. Exosome could express the common surface adhesion molecules CD63, CD81 and the surface adhesion molecules CD90, CD73, CD105, CD29, CD166 of mesenchymal stem cells. These findings indicate that exosome may be secreted by mesenchymal stem cells of human umbilical cord blood, which contains plasma membrane proteins of umbilical cord blood mesenchymal stem cells.