This study was aimed to compare the effects of acupuncture stimulation on different acupoints, which included LR14-Qimen, ST25-Tianshu, GB34-Yanglingquanand CO11-pancreas and gallbladder (auricular vagus nerve intensively distributed areas) on gallbladder pressure and Oddi sphincter function of guinea pig, in order to discuss acupuncture effect on different acupoints in the regulation of motor function of extrahepatic biliary system. A total of 20 adult guinea pigs of both sexes were randomly divided into two groups, with 10 in each group. The detection was made on changes of Oddi sphincter electromyography and gallbladder pressure before and after acupuncture. A double-hook electrode was implanted into the Oddi sphincter for the detection of Oddi sphincter electromyography. And one small artificial balloon was put into the gallbladder to measure the intrabiliary pressure. After the guinea pig was anesthetized, changes of gallbladder pressure as well as the electromyography frequency and amplitude index of Oddi sphincter of the normal guinea pig were recorded. When the guinea pig was stabilized, acupuncture was given on LR14, ST25, GB34 and CO11 for 1 min. Observations were made on acupuncture effects of gallbladder pressure and Oddi sphincter function of guinea pig. The results showed that after acupuncture on LR14 and ST25, the electromyography frequency of Oddi sphincter was obviously increased compared with before acupuncture (frequency:P < 0.01, amplitude:P < 0.05); the gallbladder pressure was obviously decreased (P < 0.05). After acupuncture on CO11 and GB34, the electromyography frequency and amplitude index of Oddi sphincter were obviously decreased compared with before acupuncture (P < 0.05); the gallbladder pressure was obviously increased (P < 0.01). It was concluded that acupuncture on LR14 and ST25 can obviously inhibit the movement of gallbladder and promote the myoelectricity release of Oddi sphincter. Acupuncturing on CO11 and GB34 can apparently promote the shrinking movement of gallbladder and inhibit the myoelectricity release of Oddi sphincter.

The physiological mechanism underlying the acupoint sensitization was evaluated systemically by using the method of electric physiology at spinal cord, medulla, and thalamus levels; the dynamic change of acupoint from the relative "silence" to the relative "activation" function was explained through the study on the dynamic process of acupoint sensitization; the biological process of the therapeutic effect of acupoint stimulation was illuminated through the research of the central mechanism underlining the dose effect relationship between the sensitive acupoint and the related brain area, thus scientific evidence for the functional link between the acupoint and internal organs as well as the nature of the acupoint were provided.
Acupuncture Points ; Acupuncture Therapy ; Animals ; Humans ; Moxibustion ; Nociceptors ; physiology ; Sensation ; Viscera ; innervation ; physiology ; Visceral Afferents ; physiology

The preliminary metabolic profile of total diterpene acid (TDA) isolated from Pseudolarix kaempferi was investigated by using in vivo and in vitro tests. Pseudolaric acid C2 (PC2) was identified as the predominant metabolite in plasma, urine, bile and feces after both oral and intravenous administrations to rats using HPLC-UV and HPLC-ESI/MS(n), and demethoxydeacetoxypseudolaric acid B (DDPB), a metabolite proposed to be the glucoside of PC2 (PC2G), as well as pseudolaric acid C (PC), pseudolaric acid A (PA), pseudolaric acid A O-beta-D glucopyranoside (PAG), pseudolaric acid B O-beta-D glucopyranoside (PBG) and deacetylpseudolaric acid A (DPA) originated from TDA could also be detected. It was demonstrated by tests that the metabolism of TDA is independent of intestinal microflora, and neither of pepsin and trypsin is in charge of metabolism of TDA, TDA is also stable in both pH environments of gastric tract and intestinal tract. The metabolites of TDA in whole blood in vitro incubation were found to be PC2, DDPB and PC2G, which demonstrated that the metabolic reaction of TDA in vivo is mainly occurred in blood and contributed to be the hydrolysis of plasma esterase to ester bond, as well as the glucosylation reaction. These results clarified the metabolic pathway of TDA for the first time, which is of great significance to the in vivo active form and acting mechanism research of P. kaempferi.
Animals ; Chromatography, High Pressure Liquid ; Diterpenes ; metabolism ; Glucosides ; metabolism ; Hydrolysis ; Mass Spectrometry ; Metabolic Networks and Pathways ; Pinaceae ; chemistry ; Rats

Objective:To investigate the influence of metastasis suppressor gene KAI1 on the proliferation,invasion and metastasis of endometrial carcinoma cell line AN3CA and HEC-1-B.Methods:The KAI1 cDNA was transfected into human endometrial carcinoma cells AN3CA and HEC-1-B via Lipofectamine 2000.The expression of KAI1 protein was ex- amined by Western blotting and flow cytometry before and after transfection.The proliferation ability of AN3CA and HEC- 1-B cells was observed by MTT assay and anchorage-independent growth assay.The changes of cell invasive ability were studied by transwell assays.Results:Stable expression of KAI1 protein was observed in AN3CA and HEC-1-B cells and on their surface after transfection with pcDNA3-KAI1 plasmid.Cells transfected with blank plasmid formed more colonies and had a larger size,with the colony forming rates being(54.2?3.1)% for AN3CA cells and(52.7?4.3)% for HEC- 1- B cells;the doubling time of AN3CA and HEC-1-B cells were 21.3 h and 20.1 h,respectively.Cells transfected with pcDNA3-KAI1 formed less colonies and had a smaller size,with the colony forming rates being(37.4?5.1)% for AN3CA cells and(32.1?3.7)% for HEC-1-B cells;the doubling time of AN3CA and HEC-1-B cells were 43.7h and 45.2 h,respectively.The cell proliferation abilities and colony-forming ability were significantly different between the two groups(P

Hepatitis associated anti-tuberculous treatment (HATT) has been a main obstacle in managing patients co-infected with Mycobacterium tuberculosis and hepatitis B virus (HBV).Therefore,we evaluated the factors related to the severity of adverse effects during HATT,especially those associated with liver failure.A retrospective study was carried out at Tongji Hospital from 2007 to 2012.Increases in serum transaminase levels of＞3,5,and 10 times the upper limit of normal (ULN) were used to define liver damage as mild,moderate,and severe,respectively.Patients with elevated total bilirubin (TBil) levels that were more than 10 times the ULN (＞171 μrnol/L) with or without decreased (＜40％) prothrombin activity (PTA) were diagnosed with liver failure.A cohort of 87 patients was analyzed.The incidence of liver damage and liver failure was 59.8％ (n=52) and 25.3％ (n=22),respectively.The following variables were correlated with the severity of hepatotoxicity:albumin (ALB) levels,PTA,platelet counts (PLT),and the use of antiretroviral therapies (P＜0.05).Hypo-proteinemia and antiretroviral therapy were significantly associated with liver failure,and high viral loads were a significant risk factor with an odds ratio (OR) of 2.066.Judicious follow-up of clinical conditions,liver function tests,and coagulation function,especially in patients with high HBV loads and hypoalbuminemia is recommended.It may be advisable to reconsider the use of antiviral drugs failure during the course of anti-tuberculous treatment of HBV infection patients to avoid the occurrence of furious liver failure.

Interleukin-6(IL-6)is a pleiotropic cytokine produced by monocytes, macrophages, T lymphocytes and other cell types. It is significantly up-regulated in the process of infection and inflammation, and is the core cytokine of the host's defense against environmental stresses(such as injury and infection). Abnormal and persistent IL-6 production is closely associated with the development of various autoimmune and inflammatory diseases. A growing number of studies have shown that IL-6 plays a critical role in ocular inflammation and angiogenesis in the conjunctiva, cornea, uvea and retina. Blockade of IL-6 ameliorates chronic and refractory intraocular inflammation. This article aims to review the role as well as the mechanism of IL-6 in ocular inflammatory diseases, attempting to have a deep and systematic understanding of the role of IL-6 in ocular inflammatory diseases.

Objective: To investigate the clinical significance of tumor associated macrophages (TAM) in multiple myeloma (MM) and the relationship with angiogenesis and immunosuppression. Methods: Seventy cases of MM patients diagnosed from August 2015 to June 2017 were enrolled in the study as experimental group, 20 cases of benign hematological diseases (13 with iron deficiency anemia and 7 with megaloblastic anemia) patients as control group. Immunohistochemical method was used to detect the expression of CD163, CD34 and VEGF in bone marrow samples, and flow cytometry was used to detect the proportion of regulatory T cell (Treg cells), ELISA was used to detect the level of IL-10, and the clinical features were analyzed. Results: ①Among the 70 patients, there were 31 males and 39 females with a median age of 65 (50~78) years old. TAM infiltration density, microvascular density (MVD), VEGF expression level, Treg ratio and IL-10 level in bone marrow samples of 70 MM patients were significantly higher than those of benign hematological diseases (P<0.05). ②In the MM group, the above indexes of the patients with disease stabilized (15 cases) were lower than those of the newly diagnosed group (35 cases) and the relapse refractory group (20 cases) (P<0.05), those of relapse refractory group were higher than those of newly diagnosed group (P>0.05). ③Of the 35 newly diagnosed MM patients, 27 completed 4 courses of treatment. In the effective group (15 cases), the TAM infiltration density after treatment was significantly lower than that before treatment, the difference was statistically significant[(20.20±7.66) vs (28.87±11.97), t=2.362, P=0.025]; while in the ineffective group of 12 cases, the difference of the TAM infiltration density before and after treatment was not statistically significant[(42.00±13.76) vs (48.25±13.59), t=1.119, P=0.275]. ④TAM infiltration density in the effective group after bortezomib treatment (21 cases) were lower than those in the non-bortezomib treatment group (18 cases)[(16.52 ±4.26) vs (19.27 ±5.82), t=1.662, P=0.170]. ⑤The TAM infiltration density in MM patients was positively correlated with MVD, VEGF expression level, Treg cell ratio and IL-10 level (P<0.001). Conclusion: The infiltration of TAM in the microenvironment of MM, which may promoting angiogenesis and inhibiting immune response, is related to the occurrence, development, therapeutic effect and drug resistance of MM.
Aged ; Female ; Humans ; Macrophages ; Male ; Middle Aged ; Multiple Myeloma ; Neoplasm Recurrence, Local ; Neovascularization, Pathologic

OBJECTIVEThrough comparison of HER2/neu oncogene detected by chromogenic in situ hybridization (CISH) and immunohistochemistry (IHC) in breast cancer, to explore the effect of CISH on detecting gene amplification of HER2.

METHODSSelected formalin-fixed paraffin-embedded breast samples whose pathological types were infiltrating ductal carcinomas (255 retrospective samples, 271 prospective samples), and these samples were detected by IHC and CISH.

RESULTS(1) In the retrospective study, CISH identified gene amplification in 91.6% of IHC score 3+ tumors (120/131) and in 56.5% of IHC score 2+ tumors (39/69), thus the concordant ratio between IHC and CISH was 81.2% (207/255). The two results showed significant correlation (P<0.01). (2) In the prospective study, the ratio of HER2 protein over expression detected by IHC was 31.7%, the ratio of HER2 gene amplification detected by CISH was 27.3%. CISH identified gene amplification in 91.4% of IHC score 3+ tumors (53/58) and in 46.4% of IHC score 2+ tumors (13/28), Concordant ratio between IHC and CISH was 89.7% (243/271). Two results showed significant correlation (P<0.01). (3) Paired CISH/FISH results were concordant in 14 of 15 cases. The remaining case was detected by FISH, but showed no HER2 gene amplification by CISH. (4) The gene amplification by CISH had a significantly reverse correlation with ER and PR expression (P<0.01).

CONCLUSIONSThe results of HER2 gene amplification detected by CISH have high concordance with the results detectd by IHC and FISH. CISH is a novel technique for detecting HER2 gene amplification.

Breast Neoplasms ; genetics ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; genetics ; metabolism ; pathology ; Female ; Gene Amplification ; Humans ; Immunohistochemistry ; methods ; In Situ Hybridization ; methods ; Prospective Studies ; Receptor, ErbB-2 ; genetics ; metabolism ; Receptors, Estrogen ; metabolism ; Receptors, Progesterone ; metabolism ; Retrospective Studies

This study was purposed to culture murine compact bone-derived mesenchymal stem cell (MSC) and analyze the immunological and trilineage differentiation potential. Tibia and femur were extracted. Bone marrow cells were flushed out and compact bone fragments were digested with collagenase. The digested cells were cultured in 6-well plates. The immunophenotype, immunosuppressive function and trilineage differentiation potential were analysed by flow cytometry, mixed lympocyte reaction and Oil red O, von Kossa and alcian blue straining, respectively. The results indicated that the pure compact bone MSC could be isolated with in 3 weeks. The resulting MSC had trilineage differentiation potential and immunosuppressive effect on mixed lymphocyte reaction. The count per minute (CPM) value in control group of BALB/c T cells cocultured with irradiated C57BL/6 T cells was (2.56 ± 0.31) × 10(4), while CPM values of mixed lymphocyte cocultured with C57BL/6 compact bone MSC at ratios of 100:1 and 10:1 were (0.47 ± 0.12) × 10(4) and (0.28 ± 0.09) × 10(4). The CPM value of control group was higher than those of MSC cocultured group (P < 0.001). Compact bone-MSC had an immunosuppressive effect on mixed lymphocyte reaction in a dose dependent manner. It is concluded that murine compact bone has rich MSC and the primary MSC is contaminated with less hematopoietic cells. Murine compact bone-MSC have immunosuppressive effect on mixed lymphocyte reaction and trilineage differentiation potential. Compact bone-MSC have promising experimental study value.
Animals ; Bone Marrow Cells ; cytology ; immunology ; Bone and Bones ; cytology ; Cells, Cultured ; Female ; Immunophenotyping ; Lymphocyte Culture Test, Mixed ; Mesenchymal Stromal Cells ; cytology ; immunology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL

Adolescent ; Cardiac Catheterization ; methods ; Child ; Child, Preschool ; Echocardiography ; Female ; Heart Septal Defects, Atrial ; diagnostic imaging ; surgery ; Humans ; Infant ; Male