Background Studies showed that macophenolic acid (MPA)down-regulates and inhibits the expression and secretion of tissue growth factor and inflammatory factor,and further impacts the proliferation and inflammation process.Pterygium is an inflammatory and proliferative lesion.Whether MPA has an inhibitory effect on pterygium is unclear.Objective This study was to investigate the antifibrotic effects of macophenolic acid on pterygium fibroblasts(PFBs) in vitro and discuss its mechanism.Methods Pterygium tissue was obtained from pterygium patient during the surgery.PFBs were cultured using explants and identified with vimentin immunohistochemisty.0,0.125,0.250,0.500,1.000 μmol/L MPA were added into the culture medium,respectively,and the cells were cultured in the medium without MPA as the control group.MTT colorimetry was used to find the optimization effective concentration of MPA and evaluate their inhibitory effect on PFBs,and BrdU fluorescence staining was used to assess the growth statue of PFBs.Expressions of nuclear factor-κB(NF-κB),p65 and inhibitor of NF-κB-α(IκB-α) in the cells were detected by Western blot.Results The cells was spindle in shape 3 days after cultured and showed the vortex and radial arrangement with the positive response to vimentin.With the increase of MPA,the proliferative value of PFBs (A560)showed gradually decline,with a significant difference among the five groups (F =42.874,P＜0.01).In addition,the proliferative value of PFBs (A560) significantly lowed as the prolong of MPA active time(F=26.038,P＜0.01).BrdU fluorescence staining showed a significant decrease of DNA synthesis of PFBs with the elevation of MPA dose among the five groups(F=175.279,P＜0.05),and the A560of PFBs DNA synthesis in different concentrations of MPA groups was lower than that of the control group (all at P＜0.05).No apoptotic and necrotic cell was found after MPA action by DAPI staining.The expression level of p65 in the PFBs was 0.886±0.072 and 1.542±0.124 in the MPA group and the control group,indicating a declined value in the MPA group(P＜0.05).However,the expression value of IκB-α in the cytoplasm PFBs was significantly higher in the MPA group compared with the control group(2.141 ±0.305 vs.1.559±0.267) (P＜0.05).Conclusions MPA has an inhibitory effect on the growth of PFBs,which probably is related to the arresting of NF-κB pathway.

OBJECTIVETo discuss the mechanism of gambogenic acid (GNA) in inducing the apoptosis of melanoma B16 cells.

METHODThe inhibitory effect of GNA on the proliferation of B16 cells was measured by the methyl thiazolyl tetrazolium (MTT) assay. The effect of GNA on B16 cells was detected by the Hoechst 33258 staining. The transmission electron microscopy was used to observe the ultra-structure changes of B16 cells. The changes in PI3K, p-PI3K, Akt, p-Akt, p-mTOR, PTEN proteins were detected by the Western blotting to discuss the molecular mechanism of GNA in inducing the apoptosis of B16 cells.

RESULTGNA showed a significant inhibitory effect in the growth and proliferation of melanoma B16 cells. The cell viability remarkably decreased with the increase of GNA concentration and the extension of the action time. The results of the Hoechst 33258 staining showed that cells processed with GNA demonstrated apparent apoptotic characteristics. Under the transmission electron microscope, B16 cells, after being treated with GNA, showed obvious morphological changes of apoptosis. The Western blot showed a time-dependent reduction in the p-PI3K and p-Akt protein expressions, with no change in p-PI3K and p-Akt protein expression quantities. The p-mTOR protein expression decreased with the extension of time, where as the PTEN protein expression showed a time-dependent increase.

CONCLUSIONGNA could inhibit the proliferation of melanoma B16 cells and induce their apoptosis within certain time and concentration ranges. Its mechanism in inducing the cell apoptosis may be related to PI3K/Akt/mTOR signaling pathways.

Animals ; Apoptosis ; drug effects ; Blotting, Western ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Melanoma ; metabolism ; pathology ; ultrastructure ; Mice ; Microscopy, Electron, Transmission ; Microscopy, Fluorescence ; PTEN Phosphohydrolase ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Signal Transduction ; drug effects ; TOR Serine-Threonine Kinases ; metabolism ; Terpenes ; pharmacology ; Xanthenes ; Xanthones ; pharmacology

AIM: To observe the effect of Baoxin capsules on hemodynamics of ischemic myocardium of dogs. METHODS: Fed Baoxin capsule for continuous three days, left anterior descending branch of the coronary artery of the dogs were ligated to observe the change of cardiac output, blood pressure and ?dp/dt max in different time of myocardial ischemia. RESULTS: Baoxin capsules increased the blood flow obviously, and increased left ventricular function simultaneously, the difference with ischemic group is significant (P

ObjectiveTo investigate the role of myocardial calcium-sensing receptor (CaSR) in a rat model of high-level spinal cord injury (SCI).MethodsEighteen healthy male SD rats weighing 250-300 g were randomly divided into 2 groups:sham operation group(group S,n ＝6) and SCI group(n ＝ 12).SCI model was induced by dropping a 10 g weight onto spinal cord (C7) in freely vertical falling along the hollow glass tube from 5 cm height.The blood samples were taken 12 and 24 h after SCI in group SCI and 12 h after SCI in group S,and serum activity of creatine kinase(CK) and MB isoenzyme of creatine kinsse(CK-MB) were measured.Then myocardium specimens were obtained for uhrastructure examination and determination of CaSR mRNA and protien expression by fluorescence quantitative RCR and Western blot.Results Serum activities of CK and CK-MB and CaSR mRNA and protein expression were higher in group SCI than in group S.Serum activity of CK and CaSR mRNA expression were higher,and serum activity of CK-MB was lower at 24 h after SCI than that at 12 h after SCI.There was no significantly difference in CaSR protein expression between the two time points in group SCI.The ultrastructure examination showed that myocardial injury was found in group SCI.ConclusionThe expression of CaSR is up-regulated after SCI in rats,which might be the mechanism of myocardial injury after SCI.

Objective To investigate the effects of salvianolate on noradrenaline(NE), angiotension-Ⅱ(Ang-Ⅱ)and tumor necrosis factor-α(TNF-α)in rats with congestive heart failure. Methods Sixty male SD rats were randomly divided into 6 groups, the normal control group(NCG), the model group, the captopril group(CAG), the low dosage of salvianolate group(LSG), the high dosage of salvianolate group(HSG), the captopril and high doseage of salvianolate group(CSG). The rats of Congestive Heart Failure were established with peritoneal injection of adriamycin except the rats in normal control group. The rats in normal control group were injected with the equal volume of normal saline once per week for 6 weeks. The medication was started in every group at the same time. Eight weeks later, the Left Ventricular Systolic Pressure(LVSP), Left Ventricular End-diastolic Pressure(LVDP), +dp/dtmax, -dp/dtmax were measured. The serum levels of NE, Ang-Ⅱ and TNF-αwere measured. Protein expressions of Ang-Ⅱ and TNF-αin cardiac muscle were detected by Western blot. Results Compared with the model group[(90.77±14.75)mmHg, (22.52±2.58)mmHg, (3 290.16± 109.61)mmHg/s, (3 114.07±112.39)mmHg/s], HSG[(114.10±13.71)mmHg, (19.97±1.14)mmHg, (3 504.97 ± 163.94)mmHg/s, (3 303.02 ± 121.98)mmHg/s ] and CSG [ (141.18 ± 15.42)mmHg, (15.58 ± 1.46)mmHg, (3 766.56±159.93)mmHg/s, (3 566.70±154.57)mmHg/s]had significant difference in LVSP, LVDP, +dp/dtmax, -dp/dtmax(P<0.05 or 0.01). Compared with the model group[(85.06±9.07)ng/ml, (180.11±25.45)pg/ml, (205.80±15.73)pg/ml], the serum levels of NE, Ang-Ⅱand TNF-αwere decreased in groups of HSG[(75.33±8.60)ng/ml, (149.21±25.39)pg/ml, (188.84±13.79)pg/ml], CAG[(71.49± 6.21)ng/ml, (139.15 ± 24.83)pg/ml, (183.73 ± 10.99)pg/ml ] and CSG [ (60.02 ± 7.38)ng/m, (110.68 ± 28.63)pg/ml, (165.93±16.22)pg/ml]in different degree(P<0.05 or 0.01), those of CSG were significantly lower than CAG(P<0.05). Compared with the model group[(1.043±0.044), (1.167±0.048)], the expression of protein in Ang-Ⅱand TNF-α were decreased in groups of HSG[(0.981±0.024), (1.069±0.055)]CAG [(0.954±0.031), (1.046±0.053)]and CSG[(0.886±0.044), (0.955±0.038)]in different degree(P<0.05 or 0.01), those of CSG were significantly lower than CAG(P<0.05). Conclusion Salvianolate can reduce the serum levels of NE, Ang-Ⅱand TNF-αin rats with congestive heart failure, decrease the expression of Ang-Ⅱand TNF-αin its cardiac muscle, improve the cardiac function.

Abstract: Objective　To investigate the pollution of paralytic shellfish poisons (PSP) in shellfish sold in Hainan Province from 2018 to 2021. Methods　From 2018 to 2021, the content of 10 paralytic shellfish poisons including saxitoxin (STX), neosaxitoxin (neoSTX), gonyautoxins 1 (GTX1), gonyautoxins 2 (GTX2), gonyautoxins 3 (GTX3), gonyautoxins 4 (GTX4), gonyautoxins 5 (GTX5), decarbamoylsaxitoxin (dcSTX), decarbamoylgonyau toxins 2 (dcGTX2) and decarbamoylgonyau toxins 3 (dcGTX3) in 7 kinds of shellfish commonly sold in 13 cities and counties in Hainan province was analyzed. Results　The detection rate of PSP in 360 shellfish samples was 10.3%. Among them, the highest detection rate of STX was 5.83%, followed by GTX2 detection rate of 4.17%; the detection rate of neoSTX and GTX3 were both 1.67%; the detection rate of GTX1 was 1.39%. None of the five PSP, GTX4, GTX5, dcSTX, dcGTX2 and dcGTX3, were detected. Four types of PSP were detected in fanscallops, two were detected in oysters, mussels and Scapharca subcrenata, only one was detected in scallops, and no toxin contamination was detected in clams and razor clams. A single sample of fanscallops detected a maximum of 4 PSP, and a single sample of oysters, scallops, mussels and Scapharca subcrenata detected a maximum of 1 PSP. The equivalence of PSP in all samples was ND-155.6 μg/kg.The annual detection rate of PSP from high to low was: 20.0% in 2020, 15.6% in 2019, 5.3% in 2018, and 2.0% in 2021, and none of the samples tested exceeded the standard. Continuously detectable STX in 2018-2020, all PSP that could be detected in 2018 were STX. In 2019, in addition to STX detected in scallops and Scapharca subcrenata, neoSTX was also detected in oysters, mussels and Scapharca subcrenata. In 2020, PSP was only detected from scallops, and GTX2 could be detected in all positive specimens, while 5 STX, 5 GTX1 and 6 GTX3 were detected. Only GTX2 detected from scallops in 2021. STX was detected in shellfish sold in 12 cities and counties, GTX2 can be detected in 10 cities and counties, neoSTX can be detected in 5 cities and counties, GTX1 and GTX2 were detected in 4 cities and counties respectively. Shellfish sold in Wenchang and Lingshui markets can detect 5 types of PSP. Conclusion　Some types of shellfish on the market in Hainan are contaminated with some kind of PSP pollution risks, and it is necessary to strengthen the supervision of PSP in marketed shellfish.

Objective To explore change of peripheral blood CD5+B cells in the systemic lupus ery- thematosus(SLE)patients and the association of this change with the disease activity,Methods The percent- age of CD5~+B ceils in peripheral blood in 57 patients with SLE and 35 healthy controls was determined by flow eytometry.Levels of anti-dsDNA,complement C3 and C4,anti-nuclear antibody(ANA),anticardiolipin anti- body(ACL),were also measured.Results The percentage of CD5~+B cell in SLE patients(2.1?0.4)% was high- er than that in normal controls [(1.5?0.4)%](P＜0.05).In active disease group,the percentage of CD5~+B cells [(2.5?0.5)%] was significantly higher than that in inactive SLE patients [(1.4?0.5)% ](P＜0.01).The percent- age of CD5~+B cell associated with anti-dsDNA,ANA,ALA positively,with C3 negatively and had no associa- tion with C4.Conclusion The percentage of CD5~+B cells in the SLE is significantly high and has correlation and some relationship with the SLE.

OBJECTIVETo find out the suitable time for collecting gallnuts. When Kaburgaia rhusicola parasailed on leaflets of Rhus potainii, the parts of the leaflets developed into gallnuts.

METHODThe regulation of development and tannin amount during whole growing stage were studied.

RESULTK. rhusicola lived in gallnuts for about 80 days. The gallnuts developed quickly during earlier stage and slowly during later stage. Gallnuts maturated from 12th, July to 30th, July, which had no relations with the size and the time for forming, but had relations with development at earlier stage. It developed quickly with earlier maturation and developed slowly with later maturation. Its tannin amount kept stable during the whole growing stage.

CONCLUSIONThe best harvesting days are those shortly after 7th, July.

Animals ; Aphids ; chemistry ; Hydrolyzable Tannins ; analysis ; Materia Medica ; analysis ; Plant Leaves ; parasitology ; Plants, Medicinal ; parasitology ; Rhus ; parasitology ; Time Factors

Objective To estimate the effect and the intracellular signal transduction pathway of insulin-like growth factor I (IGF-I) on the expression of stem cell factor(SCF) in colonic smooth-muscle cells(SMC). Methods The SMCs isolated from colon of the SD rats by enzymolysis were cultured and identified by α-actin immunocytochemical method. Colonic SMCs were cultured either with 100 μg/L of IGF-I at different time points (0, 8, 16, 24 and 48 hours) or with different concentrations (0,5,10,50,100 and 150 μg/L) of IGF-I for 16 hours. The expressions of SCF in colonic SMCs pretreated with or without speicific phosphatidylinositol 3-kinase (PI 3-kinase) inhibitor LY-294002 or mitogen-activated protein (MAP) kinase (MEK1) inhibitor PD-98059 were examined by Western blotting and quantitative reverse transeription-polymerase chain reaction and immunofluorescence. Results Low expression of SCF was found in colonic SMCs cultured in the bovine serum free medium. There was no effect of 5 or 10 μg/L of IGF- I on the expression of SCF. However, the expressions of SCF mRNA and protein were increased when stimulated with high concentrations (50,100,150 μg/L) of IGF-I. The peak expression of SCF was showed at the 16th hour after stimulating with 100 μg/L of IGF- I that was considered as the most effective concentration in vitro. The expression of SCF was not influenced by LY-294002, but was partly blocked by PD-98059. Conclusions The expression of SCF in colon SMCs may be induced by IGF-Ⅰ through MAP kinase signaling pathway.

Objective To discuss the technique and value of preoperative reformatting with three-dimensional computed tomography technique for C_2 pedicle screw track.Methods GE Light Speed 16 Pro spinal CT scans of 15 adult dry vertebrae were loaded into an imaging station (software ADW4.2).Two methods of C_2 pedicle screw techniques were analyzed through virtual screw trajectory by VR (volume rendering) and MPR (multiple planar reformatting) techniques,in method A,screw entry point was the intersection between the media-vertical and the cranial line of C_2 inferior facet joint,in method B,the screw track was from the cranial and medial quadrant of the dorsal part of C_2 inferior facet joint.Results The screw track could be observed dynamically from any plane.Two vertebrae were ob- served with smaller height in isthmus and the medial edge of the transverse foramen since no space was a- vailable for the screw.The screw trajectories data were compared between method A and method B,which showed that the angles towards the cephalad (in sagittal plane) and midline (in transverse plane) were bigger in method A than in method B (P＜0.05,0.01),but the safe screw diameter was smaller in method A than in method B (P＜0.05),and there was no difference of the screw length between the two methods(P＜0.05 ).Conclusion In this research,the individual C_2 pedicle screw entry points, screw diameter and security screw angle can be simulated,and the screw track can be observed dynami- cally to make sure if it transits the bone structure completely.Preoperative three-dimensional computed tomography reformatting for pedicle screw track is of great value in clinical and basic researches.