The effects of prostaglandin E_2 on hemodynamics and cardial function in normal rabbits and on cardioprotection in experimental acute myocardial infarction rabbits were investigated. To determine the action of prostaglandin E_2 in normal rabbits (n = 10), we measured arterial systolic pressure (ASP), arterial diastolic pressure (ADP), mean arterial pressure (MAP), heart rate (HR), left ventricular ejection time (LVET), +dp/dt max, -dp/dt max, Vc_E-50, T time after injected prostaglandin E_2 (1?g/min/kg). The results showed that ASP, ADP, MAP were decreased. HR, Vc_E-50, LVET were unchanged. To determine the effects of prostaglandin E_2 on the cardioprotection in acute myocardial infarction in rabbits (n=8), left ventricular systolic pressure (LVEP), +dp/dt max,-dp/dt max, ST of ECG, infarct size were also measured by us. The results showed that +dp/dt max, -dp/dt max were higher than control (n=9), ST lower than control, LVSP and infarct size were unchanged. Thus, prostaglandin E_2 could relax vascular smooth muscle, reduce laod of heart, decrease myocardial oxygen demands, maintain cardiac function during acute myocardial infarction.

PGE_2 reduced pulsation in concentration of 4 ng/ml and pause in 40 ng/ml for cultured rat heart muscle cells. Membrane and microfilament changes slightly in cultured heart muscle cells exposed to PGE_2 4 ng/ml for 6 min. But membrane presented marked distortion in cultured heart muscle cells exposed to PGE_2 4 ng/ml for 6 hr. These data indicated that effect of PGE_2 is time-dependent.

Objective To explore the possible functional mechanism of Er-xian Decoction and its component herbs on the expression of extrogen receptor(ER) in adrenal development rats.Methods Fifty-four development female SD rats were randomly divided into nine groups:normal control group(A),positive control group(B),Er-xian Decoction group(C),Herba Epimedii group(D),Rhizoma Curculiginis group(E),Radix Morindae Officinalis group(F),Radix Angelicae Sinensis group(G),Cortex Phellodendri group(H),Rhizoma Anemarrhenae group(I).After the rats were administrated for six days,serum were got to measure testosterone(T).Bilateral adrenal gland was weighed and embedded by paraffin.Mean optical density(MOD) were studied by the immunohistochemical method and image analyzing system.Results The adrenal coefficient was increased in group B and C,and the level of T was decreased in group C,D,E and I.ER? was mainly observed in the cytoplasm of zona glomerulosa and zona fasciculate.MOD in group C was increased while declined in group B.ER? was mainly observed in nucleus of zona glomerulosa and zona fasciculate.MOD in group C,D,E and I was less than group B.Conclusion Er-xian Decoction and its component herbs of warming Shen(Herba Epimedii,Rhizoma Curculiginis) and nourishing Yin(Rhizoma Anemarrhenae) play a phytoestrogen role by influencing the expression of ER? and ER ? in the adrenal cortex.

BACKGROUND: The association between the ingredients of perfusate and its protection on corneal endothelium is always the hot issue in ophthalmology and pharmacology.OBJECTIVE: To observe the influence of different perfusates on the structure and function of rabbit corneal endothelium during phacoemulsification.DESIGN: A randomized grouping designed and controlled animal trial.SETTINGS: Laboratory of experimental animal center of Jinzhou Medical College and the laboratory of experimental animal operation of an urban hospital.MATERIALS: The experiments were carried out in the laboratory of experimental animal center of Jinzhou Medical College and the laboratory of experimental animal operation of Jinzhou Yadong Ophthalmology Hospital from September 2004 to March 2005. Sixteen pure Japanese big-ear rabbits of 3.5 months old, clean degree, were randomly divided into four groups with 4 rabbits in each group: normal control group, saline group, shike group and balanced salt solution group. Shike was produced by Shenyang Qixing Pharmaceutical Co.,Ltd.; Balanced salt solution by Alcon Company (USA).METHODS: The rabbits were intraperitoneally anesthetized with 100 ml/L chloral hydrate (3 mL/kg), 4% oxybuprocaine hydrochloride eye drops were used for surface anesthesia of eyes, and both eyes were operated. Alcon phacoemulsification apparatus (USA) and routine microsurgical instruments were used. A 3.5-mm incision was made on sclerotic tunnel at 2 mm posterior to superior limbus of sclera, punctured into the anterior chamber, then 0.25 mL Viscoat (Alcon) was infused. Curvilinear capsulorhexis was performed with the diameter of about 5 mm. The phacoemulsification head was placed in the center to suck out the crystal nucleus and cortex, and the incision was closed after the operation. The morphology of the corneal endothelium was quantitatively determined using contact specular microscope preoperativley and 6 hours postoperatively, including the density and area of corneal endothelium,percent of hexagonal cells and the coefficient of variation. At 6 hours postoperatively, trypan blue-alizarin red active staining was performed, and the changes of slight structures of corneal endothelium were observed under light microscope.MATN OUTCOME MEASURES: ① Quantitative analysis of the forms morphology of corneal endothelium (density and area of corneal endothelium, percent of hexagonal cells and the coefficient of variation); ② Characters of forms and structures of corneal endothelium.RESULTS: All the 16 rabbits were involved in the analysis of results. ① There were no significant differences in the morphological indexes among the four groups preoperatively. ② At 6 hours postoperatively, density and areas of endothelial cells, percent of hexagonal cells and the coefficient of variation were significantly lower in the saline group,shike group and balanced salt solution group than in the normal control group (P ＜ 0.05), and no significant difference was observed between the shike group and balanced salt solution group (P ＞ 0.05). ③ The structural changes of corneal endothelium in the shike group and in balanced salt solution group were alleviated more significantly than those in the saline control group, no necrosis was observed.CONCLUSTON: In phacoemulsification, the damage of perfusate to corneal endothelium is a chemical one. Under the same surgerical conditions, domestic perfusate of shike is as effective as balanced salt solution in protecting endothelial cells.

Aim To explore the effects of tanshinone IIA on cell proliferation via G protein-coupled estrogen receptor inductive and regulative pathway in typical es-trogen receptor and G protein-coupled estrogen receptor positive T47D breast cancer cells. Methods The pro-liferation rate of T47 D cells influenced by tanshinone IIA was analyzed by MTT assay. G protein-coupled es-trogen receptor agonist G1 and GPER antagonist G15 were employed as tools. GPER SiRNA was applied to build GPER gene silence T47D cells. GPER expres-sion influenced by tanshinone IIA was measured by Western blot. Results The proliferation rates of T47D cells treated with 1 × 10 -5 mol·L-1 - 1 × 10 -7 mol· L-1 of tanshinone IIA were decreased significantly. Such effects could be attenuated by G1 or enhanced by G15 . Growth of GPER SiRNA transfected T47 D cells were significantly inhibited by 1 × 10 -5 mol·L-1 - 1 × 10 -7 mol·L-1 of tanshinone IIA treating. Result of Western blot showed that tanshinone IIA at 1 × 10 -5 mol· L-1 and 1 × 10 -6 mol · L-1 could induce de-crease of GPER protein expression in T47D cells. Conclusions Tanshinone IIA shows inhibitory effects on proliferation rate of T47 D breast cancer cells via GPER pathway. Tanshinone IIA could perform regula-tive function on GPER expression level in target cells.

This study was aimed to investigate effects of Er-Xian (EX) decoction in follicular granulosa cell apoptosis in chemotherapy-induced premature ovarian failure (POF) of rats. SD rats were randomly divided into the normal group, model group, positive group and EX decoction group. Intraperitoneal injection of cisplatin was used in the establishment of chemotherapy-induced POF rat model. Intragastric administration of corresponding medication was give to each group for four weeks. The general conditions of rats were observed. The serum contents of E2, FSH, LH and Pro were determined. The ovarian tissue morphology was observed. Granulosa cell apoptosis was analyzed by TUNEL. The expressions of BAX, Bcl-2 and VEGF protein were detected by immunohistochemistry. The results showed that in the model group, the body weight decreased with disordered estrous cycle. After treatment, the weight gained, and the estrous cycle recovered. Compared with the normal group, in the model group, E2 decreased, FSH and LH levels increased. There was no significant difference in the content of progesterone among different groups. Detection of TUNEL showed that the positive expression in the model group was increased compared with the normal group (P < 0.05). The positive expression of EX decoction group was reduced compared to the positive group (P < 0.05). Compared with the normal group, the expression of BAX was obviously increased and Bcl-2 was decreased in the model group. After EX decoction treatment, the expression of BAX was decreased and the Bcl-2 was increased compared to the model group. The expression of VEGF protein in the model group was obviously less than that in the normal group. It was concluded that cisplatin can induce follicular granulosa cell apoptosis, and cause premature ovarian function recession. EX decoction can adjust the content of different hormones in serum, alter expression of apoptosis related protein to reduce the damage of cisplatin on ovarian follicular development, in order to promote follicular development, improve and enhance the ovarian function.

Objective To evaluate the role of autophagy in the development of neuropathic pain (NP) in rats.Methods Thirty male Sprague-Dawley rats,weighing 200-220 g,were randomly divided into 3 groups (n =10 each) using a random number table:sham operation group (sham group),NP group and rapamycin (autophagy inducer) group (Rap group).Intrathecal catheter was inserted into the subarachnoid space and advanced caudally until the tip reached L4,5 segment.NP was induced by ligation of the left L5 spinal nerve (SNL) in NP and Rap groups.The L5 spinal nerve was only exposed,but not ligated in group sham.At 30 min before ligation and 2 days after operation,rapamycin 60 μg was injected intrathecally via the intrathecal catheter in Rap group,while the equal volume of vehicle (5％ dimethyl sulfoxide) was injected in sham and NP groups.The mechanical and thermal pain thresholds were measured on 1,3,5 and 7 days after ligation (T-4).The rats were sacrificed after the last measurement of pain threshold at T4.The ipsilateral L5 segment of spinal dorsal horn was removed for examination of autophagosomes (using transmission electron microscope) and for determination of the expression of LC3 Ⅱ and p62 (by Western blot) and content of IL-1β (by ELISA).Results Compared with sham group,the mechanical pain threshold at each time point and thermal pain threshold at T2-4 were significantly decreased,and the LC3 Ⅱ and p62 expression and IL-1β content were increased at T4 in group NP (P ＜ 0.05).Compared with NP group,the mechanical pain threshold at each time point,thermal pain threshold at T2-4 and LC3 Ⅱ expression at T4 were significantly increased,and the p62 expression and IL-1β content were decreased at T4 in group Rap (P ＜ 0.05).Microscopic examination showed that autophagosomes were observed in the spinal dorsal horn in NP and Rap groups,and the damage to organelles was lighter in Rap group than in NP group.Conclusion The development of NP is related to autophagic dysfunction in rats.

ObjectiveTo study the effects of tanshinoneⅡA on proliferation of cervical squamous cancer Siha cells; To discuss its possible molecular mechanism.Methods Cervical squamous cancer Siha cells were treated with different doses of tanshinoneⅡA. The effects of tanshinoneⅡA on proliferation of Siha cells were measured by MTT assay and flow cytometry analysis. The effects of tanshinoneⅡA on expression levels of phospho-extracellular regulate kinase (p-ERK) and Cyclin D in Siha cells were measured by Western blot.Results 1×10-5, 5×10-6, 1×10-6, 5×10-7 mol/L tanshinoneⅡA significantly inhibited Siha cell proliferation and such effect could be enhanced by ERα antagonist MPP and attenuated by ERβ antagonist PHTPP. 1×10-5, 5×10-6, 1×10-6 tanshinoneⅡA could significantly decrease the proliferation index of Siha cells. 1×10-5, 5×10-6, 1×10-6, 5×10-7 mol/L tanshinoneⅡA could significantly reduce the protein expression levels of p-ERK and Cyclin D of Siha cells.ConclusionTanshinoneⅡA can inhibit cervical squamous cancer Siha cell proliferation and such effect is realized via estrogen receptor pathway. TanshinoneⅡA plays anti-proliferation roles by reducing the expression levels of p-ERK and Cyclin D.

This study was aimed to observe the influence of β-sitosterol (BSS) on estrogen receptor (ER) positive the human breast cancer cell line T47D and to study its mechanisms. ER antagonist ICI182 780 was employed to observe the influence on the proliferation. Proliferations of T47D cells influenced by different concentrations of BSS were analyzed by MTT assay. Cell cycle analyses were examined by flow cytometry. The protein expression of cyclin D1 was measured by western blot analysis and cyclin D1 mRNA was quantified by real-time PCR assay. The results showed that BSS in high dose exhibited significant inhibitory effects that were partly antagonized by ICI182 780 and decreased the proliferative index on T47D cells. However, BSS in low dose obviously promoted the proliferation that was completely inhibited by ICI182 780 and increased the proliferative index on T47D cells. The mRNA and protein levels of cyclin D1 were increased in low-dose BSS. The effect was blocked by ICI182 780. It was concluded that BSS in low concentration had phytoestrogenic effect by up-regulating the expression of cyclin D1 via ER pathway.

Objective:To screen the differentially expressed genes (DEG) related to inflammatory response associated with the prognosis of colon cancer based on the bioinformatics approach, and to construct and validate a prognostic model for colon cancer.Methods:RNA sequencing and clinical data of 472 colon cancer patients and normal colon tissues of 41 healthy people were retrieved from the Cancer Genome Atlas (TCGA) database. Gene expression related to prognosis of colon cancer and clinical data were retrieved from the International Cancer Genome Consortium (ICGC) database. The retrieval time was all from the establishment of library to November 2022. A total of 200 genes associated with inflammatory response obtained from the Gene Set Enrichment Analysis (GSEA) database were compared with the RNA sequencing gene dataset of colon cancer and normal colon tissues obtained from the TCGA database, and then DEG associated with inflammatory response were obtained. The prognosis-related DEG in the TCGA database were analyzed by using Cox proportional risk model, and the inflammatory response-related DEG were intersected with the prognosis-related DEG to obtain the prognosis-related inflammatory response-related DEG. The prognostic model of colon cancer was constructed by using LASSO Cox regression. Risk scores were calculated, and colon cancer patients in the TCGA database were divided into two groups of low risk (< the median value) and high risk (≥the median value) according to the median value of risk scores. Principal component analysis (PCA) was performed on patients in both groups, and survival analysis was performed by using Kaplan-Meier method. The efficacy of risk score in predicting the overall survival (OS) of colon cancer patients in the TCGA database was analyzed based on the R software timeROC program package. Clinical data from the ICGC database were applied to externally validate the constructed prognostic model, and patients with colon cancer in the ICGC database were classified into high and low risk groups based on the median risk score of patients with colon cancer in the TCGA database. By using R software, single-sample gene set enrichment analysis (ssGESA), immunophenotyping difference analysis, immune microenvironment correlation analysis, and immune checkpoint gene difference analysis of immune cells and immune function were performed for prognosis-related inflammation response-related DEG in the TCGA database.Results:A total of 60 inflammatory response-related DEG and 12 prognosis-related DEG were obtained; and 6 prognosis-related inflammatory response-related DEG (CCL24, GP1BA, SLC4A4, SRI, SPHK1, TIMP1) were obtained by taking the intersection set. LASSO Cox regression analysis showed that a prognostic model for colon cancer was constructed based on 6 prognosis-related inflammatory response-related DEG, and the risk score was calculated as = -0.113×CCL24+0.568×GP1BA+ (-0.375)×SLC4A4+(-0.051)×SRI+0.287×SPHK1+0.345×TIMP1. PCA results showed that patients with colon cancer could be better classified into 2 clusters. The OS in the high-risk group was worse than that in the low-risk group in the TCGA database ( P < 0.001); the area of the curve (AUC) of the prognostic risk score for predicting the OS rates of 1-year, 3-year, 5-year was 0.701, 0.685, and 0.675, respectively. The OS of the low-risk group was better than that of the high-risk group in the ICGC database; AUC of the prognostic risk score for predicting the OS rates of 1-year, 2-year, 3-year was 0.760, 0.788, and 0.743, respectively. ssGSEA analysis showed that the level of immune cell infiltration in the high-risk group in the TCGA database was high, especially the scores of activated dendritic cells, macrophages, neutrophils, plasmacytoid dendritic cells, T helper cells, and follicular helper T cells in the high-risk group were higher than those in the low-risk group, while the score of helper T cells 2 (Th2) in the high-risk group was lower compared with that in the low-risk group (all P < 0.05); in terms of immune function, the high-risk group had higher scores of antigen-presenting cell (APC) co-inhibition, APC co-stimulation, immune checkpoint, human leukocyte antigen (HLA), promotion of inflammation, parainflammation, T-cell stimulation, type Ⅰ interferon (IFN) response, and type ⅡIFN response scores compared with those in the low-risk group (all P < 0.05). The results of immunophenotyping analysis showed that IFN-γ-dominant type (C2) had the highest inflammatory response score, and the differences were statistically significant when compared with trauma healing type (C1) and inflammatory response type (C3), respectively (all P < 0.05). Immune microenvironment stromal cells and immune cells were all positively correlated with prognostic risk scores ( r values were 0.35 and 0.21, respectively, both P < 0.01). The results of immune checkpoint difference analysis showed there was a statistically significant difference in programmed-death receptor ligand 1 (PD-L1) expression level between high-risk group and low-risk group ( P = 0.002), and PD-L1 expression level was positively correlated with prognostic risk score ( r = 0.23, P < 0.01). Conclusions:Inflammatory response-related genes may play an important role in tumor immunity of colon cancer and can be used in the prognostic analysis and immunotherapy of colon cancer patients.