Objective To investigate the effect and mechanism of estrodial (E2) on intracellular free calcium in the endometrial-myometrial interface (EMI) smooth muscle cells from uteri with adenomyosis.MethodsFrom March 2011 to October 2011,16 uterus specimens were collected from patients with adenomyosis undergoing hysterectomy in Beijing Obstetrics and Gynecology Hospital,which included 9 proliferative endometrium and 7 secretory endometrium.EMI smooth muscle cells from the uterus were cultured and loaded with calcium ion ( Ca2 + ) fluorescent probe fluo-4/AM.The labeled cells were stimulated with the various concentration of E2 ( 1 × 102,1 × 103,1 × 104,1 × 105 pmol/L,respectively),then the changes of intracellular Ca2+ fluorescence intensity were measured by laser scanning microscopy.The most suitable concentration of E2 was selected,and the reaction difference between the EMI smooth muscle cells of two menstrual phases were also investigated; The changes of intracellular Ca2 + fluorescence intensity were detected proliferative and secretory smooth muscle cells in E2 conjugated to bovine serum albumin (17β-E2-BSA) group,cycloheximide (CHX) group,fulvestrant (ICI182780) group and pertussis toxin (PTX) group.Results ( 1 ) The cell viability of primary cultured EMI smooth muscle cells was well at 24 hours culture.(2) 1 × 102 - 1 × 105 pmol/L E2 can rapidly increase the intracellular Ca2+ fluorescence intensity within 1 min ( P ＜ 0.01 ) ;The increased amplitudes caused by 1 × 104 pmol/L and 1 × 105 pmoL/L E2 were the most significant,but there was no significant difference between them (P ＞0.05).1 × 104 pmol/L was the most suitable concentration.( 3 ) With the 1 × 104 pmol/L E2,the Ca2+ fluorescence intensity changes showed no significant difference between the EMI smooth muscle cells from the proliferative phase and secretory phase uterus (P ＞ 0.05 ).The Ca2+ fluorescence intensity changes were 646 ± 32 in 17β-E2-BSA group and 602 ±31 in CHX group,when compared with 513 ±26 and 617 ±35 in respective control group,no significant difference was observed (P ＞ 0.05 ).The increased amplitude of 188 ± 20 in the PTX group and 302 ± 11 in ICI182780 group exhibited significant difference with 632 ± 33 and 635 ± 24 in respective control group ( P ＜ 0.01 ).Conclusion E2 could increase the intracellular Ca2 + of EMI through a membrane receptor dependent and nongenomic mechanism of action.

The mechanism of biological actions of quercetin was studied by using metabolomic method and biomolecular network. HPLC-MS was used to analyze the serum metabolome in rats of blank group and quercetin administration group rats, and MS data were processed by MATLAB software. With multivariate statistical analysis of serum metabolite profiles, a clear separation among blank group and quercetin administration group was achieved, potential biomarkers were selected according to the parameters of variable importance in the projection (VIP) and identified according to MS information and database retrieval. Four compounds, related enzymes, action targets and metabolic pathways had been confirmed, namely retinoic acid and RARbeta, arachidonate and COX-2, 3, 5-diodotyrosine and TPO, uridine diphosphate glucose and PDEs. The mechanism of quercetin enhancing ability of retinoic acid on the induction of RARbeta, activating TPO, using as COX-2 and PDEs inhibitor was approved by biomolecular network and related literatures. In this study, a mechanism of multiple biological actions of quercetin was evaluated at the level of the biomolecular network, metabolomics and biomolecular network can be used to investigate the biological effects mechanism of quercetin, which provided a new method to further revealing mechanism of drug action.
Animals ; Antioxidants ; pharmacology ; Biomarkers ; blood ; Chromatography, Liquid ; Mass Spectrometry ; Metabolic Networks and Pathways ; Metabolome ; Metabolomics ; Multivariate Analysis ; Quercetin ; pharmacology ; Rats

OBJECTIVETo compare the effect and differences sex the influence of hormone levels of perimenopau-sal syndrome patients between manual acupuncture and electroacupuncture (EA).

METHODSA total of 50 cases with perimenopausal syndrome were randomly assigned into an manual acupuncture group (27 cases) and an EA group (23 cases), and 1 case dropped in the EA group. The acupoints in the two groups were Guanyuan (CV 4), Zigong (EX-CA 1), Tianshu (ST 25), and Sanyinjiao (SP 6). Acupuncture with 3-time small and even manipulation of lifting, thrusting and twirling was used in the acupuncture group, once 10 min. EA with sparse-dense wave and 10 Hz/50 Hz was applied in the EA group for 30 min. The treatments in the two groups were for continuous 8 weeks (24 times in total), once the other day, 3 times a week. The scores of 24-hour hot flashes even, menopausal rating scale (MRS) and menopause-specific quality of life questionnaire (MENQOL) were recorded before treatment and after 4-week and 8-week treatment, as well as 12 and 24 weeks after treatment. Serum sex hormone levels were tested before and after 8-week treatment as well as 12 weeks after treatment, including serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estracliol (E).

RESULTSCompared with those before treatment, the 24-hour hot flashes even score, MRS and MENQOL scores were significantly lower after 4-week and 8-week treatments, 12 and 24 weeks after treatment (all<0.05). All the above scores after 8-week treatment were lower than those after 4-week treatment (all<0.05); and the scores 12 and 24 weeks after treatment were lower than those after 4-week and 8-week treatments (all<0.05); all the scores after treatment were not significantly different at any time between the two groups (all>0.05). Compared with those before treatment, serum FSH and Eapparently improved in the two groups after 8-week treatment and 12 weeks after treatment (all<0.05). LH levels did not significantly change in the two groups (all>0.05). All the serum sex hormone levels showed no significant difference between the two groups (all>0.05).

CONCLUSIONSBoth acupuncture and EA can improve perimenopausal symptoms and serum sex hormone. The effects are similar.

LC-MS/MS method was used to simultaneously determine anti-oxidative active catechins EGCG, ECG, EGC and EC in plasma of rats treated with tea polyphenols (TP). The integrated plasma concentration (C') of TP was calculated by means of self-defined weighing coefficient based on percent AUC of individual components, thereby assessing integrated pharmacokinetic (PK) parameters of TP via log C'-T curve. The anti-free radical effects of TP were estimated using inhibitory rate of drug-containing serum collected at different times from rats against in vitro lipid peroxidation of mouse liver homogenate. The obtained E-T curves were used to calculate anti-free radical pharmacodynamic (PD) parameters of TP. E-logC and E-log C' plots and linear regression were carried out in order to obtain the correlation coefficient (R2). The results indicated that the log C'-T curves of TP, which could be best described by three-compartment model, corresponded to elimination rule of iv administration of drugs. The integrated PK parameters showed that TP was distributed in body rapidly and widely, and eliminated from deep compartment slowly. From comparison of R2 values and consistence of C'-T course and E-T course, it was evident that TP integrated PK behaviors correlated much better with its PD behaviors than individual active components, and thus demonstrated that integrated PK parameters could characterize to maximal extent holistic disposition of Chinese herbal drugs and reflect residence properties of holistic effective substances in biological body.
Animals ; Antioxidants ; pharmacokinetics ; pharmacology ; Area Under Curve ; Catechin ; analogs & derivatives ; pharmacokinetics ; Chromatography, Liquid ; Free Radical Scavengers ; blood ; pharmacokinetics ; pharmacology ; Free Radicals ; metabolism ; Injections, Intravenous ; Lipid Peroxidation ; drug effects ; Male ; Mice ; Polyphenols ; blood ; pharmacokinetics ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; Tea ; chemistry

OBJECTIVETo study the role of urotension-II in serum and bronchoalveolar lavage fluid (BALF) in the process of airway remodelling in asthmatic rats.

METHODSThirty-two male Sprague-Dawley (SD) rats were randomly divided into normal control and 2-week, 4-week and 8-week asthmatic groups (OVA inhalation of 2, 4 and 8 weeks respectively). Rats were sensitized and challenged by OVA to establish a model of asthma. The bronchial wall thickness and the airway smooth muscle thickness were measured by image analysis system. The urotension-II contents in serum and BALF were determined using ELISA.

RESULTSThe bronchial wall thickness and the airway smooth muscle thickness in the three asthmatic groups significantly increased compared with those in the normal control group (P<0.01). The urotension-II contents in serum and BALF in the three asthmatic groups also increased significantly compared with those in the normal control group (P<0.01). The urotension-II contents in serum and BALF in the 8-week asthmatic group were the highest, followed by the 4-week and the 2-week asthmatic groups (P<0.01). BALF urotension-II contents were positively correlated with the bronchial wall thickness and the airway smooth muscle thickness as well as serum U-II contents in the four groups.

CONCLUSIONSThe urotension-II contents in serum and BALF in the process of airway remodeling increase in asthmatic rats. The changes in serum and BALF urotension-II contents may be associated with airway remodeling in asthmatic rats.

Airway Remodeling ; Animals ; Asthma ; metabolism ; pathology ; Bronchi ; pathology ; Bronchoalveolar Lavage Fluid ; chemistry ; Male ; Muscle, Smooth ; pathology ; Rats ; Rats, Sprague-Dawley ; Urotensins ; analysis ; blood

OBJECTIVE15-lipoxygenase (15-LO) is a prooxidant enzyme which is expressed in asthmatic lungs leading to formation of pro- and anti-inflammatory mediators. Gene expression profiling studies show the association between 15-LO and allergic asthma. This study was designed to observe the expression of 15-LO in lungs of asthmatic rats and examine the effects of dexamethasone on 15-lipoxygenase expression.

METHODSTwenty-seven male Sprague-Dawley (SD) rats were randomly divided into three groups: control, asthma and dexamethasone (DXM) intervention. An asthma model was prepared by sensitization and challenging with ovalbumin. The production of 15-LO in lung tissue homogenates was measured using ELISA.The expression of 15-LO mRNA in lungs was determined by reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSThe levels of 15-LO mRNA and protein in the asthma group (0.51 ± 0.14 and 2080 ± 73 μg/mL, respectively) were lower than those in the control group (0.76 ± 0.15 and 2472 ± 106 μg/mL, respectively; P<0.01). DXM intervention increased significantly the levels of 15-LO mRNA and protein (1.02 ± 0.34 and 2562 ± 218 μg/mL) compared with the asthma group (P<0.01).

CONCLUSIONSThe production of 15-LO in lung tissues is reduced in asthmatic rats. DXM can increase the expression of 15-LO in lung tissues and thus might provide anti-inflammatory effects in asthmatic rats.

Animals ; Anti-Inflammatory Agents ; pharmacology ; Arachidonate 15-Lipoxygenase ; analysis ; genetics ; Asthma ; drug therapy ; enzymology ; Dexamethasone ; pharmacology ; Lung ; enzymology ; pathology ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the clinical effects of Achillon for the treatment of acute Achilles tendon rupture (AATR).

METHODSFrom April 2009 to April 2010, 19 patients with AATR who were treated with Achillon were retrospectively analyzed. There were 17 males and 2 females, with an average age of 40.2 years (30 to 58 years). There were 9 cases of sports injury, and 2 case of fall injury. The time from injury to surgery ranged from 0 to 8 days (2.2 days on average). The results of Thompson test and single heel rise test were positive in 19 cases. Clinical data were assessed with the patient satisfaction and the AOFAS hindfoot score during follow-up.

RESULTSAll the patients were followed up, and the duration ranged from 12 to 28 months (19.9 months on average). The average operation time was 41 minutes. There were no wound infections, recurrent rupture, or sural nerve complications. At the latest follow-up, 18 patients were totally satisfied with the surgical result, 1 patient feel generally due to mild pain when running. None of the patients were dissatisfied with the final results the latest follow-up. At the latest follow-up, the AOFAS score was 98.42 +/- 3.29 (89 to 100). All the patients regained normal range of motion and were able to resume their previous activities at six months after operation, with a high rate of satisfaction. Average decreased of mid-calf circumference was (0.82 +/- 0.85) cm (ranged from 0 to 3 cm).

CONCLUSIONTreatment with Achillon is safe, effective for AATR with low incidence of complications and early active rehabilitation can be carried out. It is a good method to treat AATR.

Achilles Tendon ; injuries ; surgery ; Acute Disease ; Adult ; Female ; Humans ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; instrumentation ; Retrospective Studies ; Rupture ; Suture Techniques ; instrumentation ; Tendon Injuries ; surgery

OBJECTIVETo study soft tissue changes observed through musculoskeletal ultrasound (MSUS) in the treatment of knee osteoarthritis with needle-knife, so as to provide MSUS basis for needle-knife in the treatment of knee osteoarthritis.

METHODSForty patients with knee osteoarthritis treated in the Third Affiliated Hospital of Beijing University of Chinese Medicine from December 2011 to December 2012 were selected according to inclusion and exclusion criteria. All the patients were treated with needle-knife release method. The VAS scores and knee joint circumference were recorded before treatment and 2 weeks after treatment. The changes of knee joint hydrops articuli and joint synovial thickness were measured through MSUS.

RESULTSThe knee pain index was 6.850 +/- 1.417 before treatment and 2.790 +/- 1.299 after treatment;the index after treatment was lower than that of before treatment. The knee joint circumference was 407.320 +/- 45.151 mm before treatment and 391.240 +/- 41.129 mm after treatment; the knee joint circumference decreased after treatment. The amount of hydrops articuli observed by musculoskeletal ultrasound showed that 47 knees were cured, 19 knees improved and 2 knees failed. The synovial membrane thickness: 43 knees cured, 17 knees improved and 8 knees failed.

CONCLUSIONThe hydrops articuli and synovial thickness of knee joint of patients with knee osteoarthritis observed under the MSUS is consistent with the main symptoms and signs, which suggests that MSUS observation on soft tissue changes before and after needle knife in the treatment of knee osteoarthritis with high reliability.

Adult ; Aged ; Female ; Humans ; Knee Joint ; diagnostic imaging ; pathology ; Male ; Middle Aged ; Needles ; Osteoarthritis, Knee ; complications ; diagnostic imaging ; pathology ; surgery ; Pain ; complications ; Synovial Membrane ; pathology ; Treatment Outcome ; Ultrasonography

AIM To optimize the two-phase (ethanol-dipotassium phosphate) aqueous extraction for Astragali Radix by central composite design-response surface method.METHODS With ethanol concentration,dipotassium hydrogen phosphate solution concentration and Astragali Radix feed ratio as influencing factors,together with extraction rates of total flavonoids and total saponins as evaluation indices,the technology optimization was accomplished by central composite design-response surface method.RESULTS The optimal conditions were determined to be 27.97％ for ethanol concentration,22.03％ for dipotassium hydrogen phosphate solution concentration,and 1 ∶ 58.85 for Astragali Radix feed ratio.The extraction rate of total flavonoids reached 74.13％ (concentrated in the upper layer),while that of total saponins reached 81.34％ (concentrated in the lower layer).CONCLUSION With good predictability,this simple and accurate method can be used for the extraction of total flavonoids and total saponins in Astragali Radix.

Objective To evaluate the effect of serum chromium on oral cancer after adjusting the covarite between groups based on propensity score matching (PSM). Methods We performed a case-control study in 395 cases of newly diagnosed primary oral cancer from the First Affiliated Hospital of Fujian Medical University and 1 240 controls from the same community from January 2010 to February 2018. Using the PSM to select 309 controls randomly which were matched with the cases by 1 ∶1 matching. Conditional Logistic regression model was used to explore the association between chromium and oral cancer. Results The level of serum chromium was 178.91 (121.83-284.19) μg/L in the case group, which was lower than 324.27 (264.82-397.69) μg/L in control group, the difference was statistically significant (P＜0.001). Dose-response analysis showed that the risk of oral cancer gradually decreased with the increase of serum chromium, which presented a negative correlation. There was a negative correlation between serum chromium level and the risk for oral cancer by conditional Logistic regression,the aOR of serum chromium in the Q2, Q3 and Q4 compared with the Q1 were 0.14 (0.08-0.26), 0.15 (0.08-0.28) and 0.10 (0.05-0.20),with significant trend (Ptrend＜0.001). Stratified analysis showed the negative correlation between serum chromium and oral cancer by smoking, drinking tea, not drinking alcohol status and fish, fruits and green vegetables intake frequencies. Conclusions The high level of serum chromium is a protective factor for the incidence of oral cancer, and the higher of serum chromium, the lower risk of developing oral cancer.