OBJECTIVE:To establish HPLC method for the content determination of oleanolic acid and ursolic acid in Prunella vulgris with ?-CD as additive agent of mobile phase.METHODS:The sample was Prunella vulgaris.The separation was performed on a Tigerkin ODS3(250 mm?4.6 mm,5 ?m) column with mobile phase consisted of methanol-10 mmol?L-1 ?-CD 1% ammonium acetate(86:14) at flow rate of 1.0 mL?min-1.The column temperature was set at 40 ℃ and UV detection wavelength was 210 nm.RESULTS:The linear range of oleanolic acid and ursolic acid were 0.5～5.0 ?g(r=0.999 2) and 1.0～10.0 ?g(r=0.999 8) respectively.High,medium and low average recovery of oleanolic acid were 97.3%(RSD=1.04%),98.0%(RSD=0.87%),101.6%(RSD=1.65%),respectively.Those of ursolic acid were 97.8%(RSD=1.17%),98.7%(RSD=0.93%),100.4%(RSD=1.72%),respectively.CONCLUSION:The new method is simple,accurate and reproducible for the quality control of P.vulgaris and simultaneous determination of oleanolic acid and ursolic acid.

Objective:To establish adoptive transfer model mice of type 1 diabetes for investigating the pathogenesis of type 1 diabetes.Methods:16 NOD.Scid mice were randomly divided into two groups.One was injected i.p. with splenocytes which were obtained from a diabetic NOD mouse and the other was injected with splenocytes obtained from a normal one.The blood glucose levels and weight were detected daily.The mice were killed when they had significant clinic appearance or 10 weeks after cell injection,followed by pathologic studies and cytokine assays.Results:The incidence of adoption group was 8/8 whereas that of control group was 0/8.The pathology scores of 2 groups were 2.5?0.2,0 separately.The amounts of IL-2,IL-10,IFN-? were 60.7 pg/ml,15.5 pg/ml and 20.2 ng/ml respectively in adoption group,and those in control group were 2.4 pg/ml,17.5 pg/ml,3.2 ng/ml.Conclusion:Model mice of type 1 diabetes can be established in NOD.Scid mice during the 5-10 week after adoptive transfer of splenocytes of diabetic NOD mice.

OBJECTIVE:To estabish a RP-HPLC method for the content determination of forsythin of Yinqiao jiedu pills.METHODS:The determination was performed using SHIM-PACK VP-ODS column(150 mm ? 4.6 mm,5 ?m) with column temperature at 30 ℃.The mobile phase was acetonitril-water(25∶75) with flow rate at 1.0 mL?min-1.The detective wavelength was set at 277 nm.RESULTS:The linear range of forsythin was 0.453～3.481 ?g(r=0.999 8).The mean recovery was 99.50%(RSD=1.15%,n=6).CONCLUSION:The method was simple,rapid,accurate and suitable for the quality control of Yinqiao jiedu pills.

Objective To investigate the inhibitory effects of CIK on the proliferation of Lewis lung carcinoma cell line and the growth of transplanted Lewis lung carcinoma in C57BL/6N mice in vivo.Methods CIK cells were induced by culturing PBMC with regular method.The proliferation of Lewis lung carcinoma cells was measured by MTT assay.Uhramicrostrueture of Lewis lung carcinoma cells was observed under a transmission electron microscope.Flowcytometric analysis was used to detect cell apoptosis.Ultrastructural observation expressions of FasL were individually determined by MTT and immunocytochemistry(ICC) analysis.Results Electron microscopic observations sbowed that CIK cells could induce the hepatoma cells to apoptosis.Flow cytometric analysis demonstrated that apoptosis cells of Lewis lung carcinoma were increased in CIK group compared with those in the control group.FasL expression on CIK increased.Cytotoxieity was blocked after addition of anti-FasmAb.Conclusion CIK has inhibitory effect on Lewis lung carcinoma cells both in vitro and in vivo.CIK cells can induce the apoptosis of Lewis lung carcinoraa cells.and Fas/FasL pathway plays an important role in apoptosis of Lewis lung carcinoma cells by CIK.

ObjectiveTo evaluate the safety and feasibility of the standardized management model of anesthesia for painless digestive endoscopy.MethodsData of 17 100 patients who underwent painless endoscopy were reviewed for severe adverse reaction and complications.The model included anaesthetist-directed appointment,nurse assistance during operation,and postoperative nurse observation.Eight hundred cases (200 of gastric endoscopy,intestinal endoscopy,EUS and ERCP respectively) were randomly selected and analyzed for times of endoscopic diagnosis,anesthesia,wakening and discharge,and complications.ResultsOf the 17 100 cases,severe complications occurred in 10 (0.058％ ),including 3 apnea,one respiratory obstruction due to opisthognathism and glossoptosis,five larygneal spasm and 1 reflux inspiration.There was no anesthesia or endoscopy related death.Study of 800 cases showed intraoperative MAP,HR increase or decrease over 30％ of the baseline,the incidence of SpO2 ＜ 95％ were 6.0％ ～ 25.0％,3.0％ ～8.5％,≤2.0％,respectively.The rate of lethe,good quality of sleep were 99％ ～ 100％ and 98.0％ ～100.0％,respectively.The rates of cough,body movement and myoclonic were 0.5％ ～4.5％,5.5％ ～11.5％,and 1.5％ ～ 3.5％,respectively.Rates of nausea and vomiting,excitement,restlessness and dizziness were lower than 4％.ConclusionThe standardized management model,feasible,safe and effective,is able to facilitate anesthetic efficacy and reduce complications.

BACKGROUND/AIMS: To establish an animal model of laryngopharyngeal reflux (LPR) and study the effect of LPR on the laryngopharyngeal mucosal ultrastructure. METHODS: Ten Bama minipigs were randomly divided into control group and stent group. Every pig underwent endoscope, and baseline pH was monitored for 4 hours at laryngopharynx and distal esophagus, then specimens from laryngopharyngeal mucosa were biopsied. For the control group, these procedures were repeated on the 14th day. In the stent group, a custom-designed esophageal stent suit was implanted into esophagus, laryngopharyngeal and distal esophageal pH monitoring lasted for 2 hours, then stent suit was removed 3 days later. At last, the same procedures were done as the control group on the 14th day. Specimens were observed under transmission electron microscope to measure the intercellular space and desmosome number. RESULTS: In the control group, there was no laryngopharyngeal reflux on the first day and 14th day. Before the stent was implanted, there was also no laryngopharyngeal reflux in the stent group. In both 2 hours and 14 days after stent implantation, the num -ber of reflux, reflux time, and percentage time of pH < 4.0 were significantly increased (P < 0.05) in the stent group. There was no difference in intercellular space and desmosomes in the control group between baseline and 14th day. In the stent group, intercellular space of laryngopharyngeal mucosa was significantly increased (0.37 mum vs 0.96 mum, P = 0.008), and the number of desmosomes was significantly decreased (20.25 vs 9.5, P = 0.003). CONCLUSIONS: A Bama minipig model of LPR was established by implanting a custom-designed stent suit. LPR might destroy the laryngophar yngeal mucosal barrier.
Desmosomes ; Endoscopes ; Esophageal pH Monitoring ; Esophagus ; Extracellular Space ; Hydrogen-Ion Concentration ; Hypopharynx ; Laryngopharyngeal Reflux* ; Models, Animal ; Mucous Membrane ; Stents ; Swine, Miniature*

The internal limiting membrane located at vitreoretinal interface is formed by the contiguous basement membranes of Müller cells.Nowadays, vitrectomy combined with internal limiting membrane peeling has been widely used in many operations involving macular area.Although its clinical efficacy and safety have been demonstrated, it lacks the necessary histological support.At the same time, many studies have shown that the internal limiting membrane plays different roles in the occurrence of different diseases.Current studies have found that the proliferation of inflammatory cells, glial cells and vitreous cells leads to the physiological dysfunction of the vitreoretinal interface, and the internal limiting membrane can also become a scaffold for the proliferation of myofibroblasts, which will lead to the occurrence of macular diseases.This article reviewed the histological research of internal limiting membrane in terms of diabetic retinopathy, idiopathic macular hole and idiopathic macular epiretinal membrane, hoping to better understand the internal limiting membrane under pathological conditions and to confirm the safety and necessity of internal limiting membrane peeling from ultrastructure.

OBJECTIVETo investigate the changes in local endometrial contents of estrogen receptors (ER) and progesterone receptors (PR) after insertion of levonorgestrel-releasing intrauterine system (LNG-IUS) and evaluate the efficacy of LNG-IUS in treating endometrial hyperplasia.

METHODSThe endometrial histological changes were observed in 25 anovulatory women with dysfunctional uterine bleeding after insertion of LNG-IUS, and the contents of ERs and PRs in the endometrium were measured by immunohistochemistry.

RESULTSThe endometrial proliferation activity was obviously inhibited 6 months after LNG-IUS insertion with decreased endometrial glands, glandular dysplasia and decidualization of interstitial cells. The positive cell rate for ERs and PRs in the glandular epithelial and interstitial cells were significantly reduced after LNG-IUS insertion.

CONCLUSIONSLNG-IUS can reduce ER and PR expressions in the endometrium and inhibit endometrial proliferation, and therefore can be effective in treating simple and complex endometrial hyperplasia.

Adult ; Endometrial Hyperplasia ; metabolism ; pathology ; Endometrium ; pathology ; Female ; Gene Expression Regulation ; drug effects ; Humans ; Intrauterine Devices, Medicated ; Levonorgestrel ; administration & dosage ; pharmacology ; Middle Aged ; Receptors, Estrogen ; metabolism ; Receptors, Progesterone ; metabolism

OBJECTIVETo investigate the roles of MMPs-9, TIMP-1 and TIMP-2 in cesarean section scar healing.

METHODSThe expressions of the MMPs-9, TIMP-1 and TIMP-2 were detected by EnVision immunohistochemistry in 22 pregnant women with serious complications of the uterine scar, including 8 with early caesarean scar pregnancy (CSP) and 14 with full-term pregnancy undergoing hysterectomy for placenta previa or implanted placenta. Thirty-eight full-term pregnant women without serious complications of the uterine scar and 32 normal full-term pregnant women served as the control I and control II groups, respectively.

RESULTSThe expressions of MMPs-9 and TIMP-1 differed significantly between the 3 groups (P<0.05), whereas TIMP-2 did not (P>0.05). Spearman rank correlation analysis showed that the expression of MMPs-9 in the uterine scar tissues was positively correlated with poor uterine scar healing with the correlation coefficients of 0.309 and 0.643. An increased severity of poor healing scar was associated with a significantly increased expression of MMPs-9 (P<0.05).

CONCLUSIONThe imbalanced expressions of MMPs-9 and TIMP-1 in injury repair can be related to poor uterine scar healing and CSP.

Adult ; Cesarean Section ; adverse effects ; Cicatrix ; etiology ; metabolism ; Female ; Humans ; Matrix Metalloproteinase 9 ; metabolism ; Placenta Previa ; surgery ; Pregnancy ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism ; Tissue Inhibitor of Metalloproteinase-2 ; metabolism ; Uterus ; pathology ; Wound Healing ; Young Adult

Cinnamaldehyde was shown to have significant anti-inflammatory and anti-pyretic actions in studies from both others' and our lab. Prostaglandin E2 (PGE2) plays a key role in generation of these pathological states, while PGE, synthase-1 (mPGES-1) is one of crucial biological elements in the process of PGE2 production. And as a downstream inducible terminal prostaglandin synthase of COX-2, mPGES-1 is now regarded as a more promising novel drug target than COX-2 and is attracting more and more attention from both academia and pharmaceutical industry. The purpose of present study was to further investigate the anti-inflammatory and antipyretic molecular mechanisms of cinnamaldehyde based on the mouse macrophage cell line RAW264. 7 in vitro. The PGE2 was identified by using the method of enzyme-linked immunosorbent assay (ELISA) and the expression of COX-2 and mPGES-1 at mRNA and protein levels was detected by the Real-time PCR and Western blotting methods respectively. The experimental results suggested that cinnamaldehyde could evidently reverse the increased production of PGE2induced by IL-1beta. Moreover, the up-regulated expression levels of mPGES-1 and COX-2 were significatly decreased. Together, these results provide compelling evidence that the down-regulated actions to both the production of PGE2 as well as the expression of mPGES-I might account for, at least in part, the anti-inflammatory and anti-pyretic effects of cinnamaldehyde.
Acrolein ; analogs & derivatives ; pharmacology ; Animals ; Blotting, Western ; Cell Line ; Dinoprostone ; metabolism ; Interleukin-1beta ; pharmacology ; Intramolecular Oxidoreductases ; metabolism ; Macrophages ; drug effects ; metabolism ; Mice ; Prostaglandin-E Synthases ; Real-Time Polymerase Chain Reaction