【Objective】 To investigate the effects of chlorogenic acid on the proliferation, migration and invasion of renal carcinoma A498 and 769-P cells and the possible molecular mechanism. 【Methods】 Human renal carcinoma A498 and 769-P cells were divided into control group and chlorogenic acid group (2 μL,1 μmol/L) and cultured for 72 h. The cell proliferation, invasion and migration were detected with MTT assay, Transwell assay and scratch test, respectively. The expressions of IL-1β, EPAS-1 and AKT/P65 signaling pathway related proteins were detected with ELISA, qRT-PCR and Western blot, respectively. 【Results】 Chlorogenic acid inhibited the proliferation, invasion and migration of renal carcinoma A498 and 769-P cells, and reduced the IL-1β level in the cell supernatant. Anti-IL-1β reduced the protein and mRNA expressions of EPAS-1, p-AKT and p-P65. Compared with the control group, the chlorogenic acid group had reduced mRNA and protein expressions of EPAS-1, p-AKT and p-P65 (P<0.05). 【Conclusion】 Chlorogenic acid can inhibit the invasion and metastasis of renal carcinoma cells, and its mechanism may be related to inhibiting the secretion of IL-1β, thereby inhibiting the AKT/P65/EPAS-1 pathway.

Objective:To evaluate the efficacy of Chaichuan Tongren Mixture supplemented with hysteroscopic recanalization in the treatment of tubal obstructive infertility of liver depression and qi stagnation type.Methods:A total of 60 patients with tubal obstructive infertility of liver depression and qi stagnation type who met the inclusion criteria were selected, between August 2018 and June 2020, and they were divided into the observation group with 31 cases and the control group with 29 cases, according to the random number table method. The control group was treated with hysteroscopic recanalization, while the observation group was given Chaichuan Tongren Mixture on the basis of the control group. Both groups were treated for 4 months and followed up for 12 months. TCM syndromes were scored before and after treatment, and prostaglandin E 2 (PGE 2) was detected by fully automatic biochemical analyzer and levels of TNF-α, CRP and IL-6 were measured by ELISA. The tubal patency of patients after treatment was observed, the pregnancy status was recorded after the end of follow-up and the clinical efficacy was evaluated. Results:The total effective rate was 96.77% in the observation group and that in the control group was 75.86% ( χ2=4.01, P<0.01). The scores of breast distending pain, irregular menstruation, dark purple menstrual flow and lumbosacral pain in the observation group after treatment were significantly lower than those in the control group ( t values were 17.69, 21.67, 20.89, 14.67, 18.20, respectively, all Ps<0.001). After treatment, the levels of serum PGE 2, TNF-α, CRP, and IL-6 were significantly lower in the observation group than those in the control group [(65.31±6.73) ng/L vs. (87.10±8.85) ng/L, t=10.78; (6.90±0.71) ng/L vs. (11.35±1.23) ng/L, t=17.30; (2.47±0.25) mg/L vs. (5.10±0.52) mg/L, t=25.23; (12.38±1.26) ng/L vs. (30.16±3.15) ng/L, t=29.05] ( P<0.01). After treatment, there was statistical significance in the total effective rate of tubal patency of 96.8% (30/31) in the observation group compared to 79.3% (23/29) in the control group ( χ2=4.43, P<0.01). At the end of follow-up, the pregnancy rate was 71.0% (22/31) in the observation group and 44.8% (13/29) in the control group, and the difference was statistically significant ( χ2=4.21, P<0.01). Conclusion:The Chaichuan Tongren Mixture supplemented with hysteroscopic recanalization can reduce the levels of inflammatory mediators and increase the pregnancy rate of patients with tubal obstructive infertility of liver depression and qi stagnation type.

Objective Polyinosinic-polycytidylic acid (poly IC) plays an important role in the central nervous system damage and repair.This study was to investigate the effect of poly IC on inflammatory response after spinal cord ischemia-reperfusion injury (SCIRI) in rats.Methods A total of 72 healthy adult male SD rats were equally randomized into a sham-operation, an ischemia-reperfusion (IR), and a poly IC group.The abdominal cavities of the rats were cut open and closed again in the sham-operation group and SCIRI models were established in the IR and poly IC groups by clamping the abdominal aorta, followed by reperfusion 60 minutes later and intraperitoneal injection of saline (0.1 mL) and poly IC (1.25 μg/g), respectively.At 6, 24, and 48 hours after modeling, BBB scores were obtained and the contents of TNFα, IL-1β and IFN-β were measured by ELISA.At 48 hours, the expressions of NF-κB and IL-10 were determined by immunohistochemistry, the area of ischemic necrosis in the spinal cord tissue was calculated by TTC staining, and its morphological changes were observed under the optical microscope.Results At 48 hours after modeling, the BBB scores were significantly lower in the IR and poly IC groups than in the sham-operation group (3.80±0.75 and 9.40±0.49 vs 20.00±0.00, P<0.01), though higher in the poly IC than in the IR group (P<0.01).The rats of the IR group showed extensive degenerated neurons in the gray substance of the spinal cord, with scattered foci of bleeding and blood coagulation, while those of the poly IC group exhibited fewer necrotic neurons and basically normal nuclear morphology, though with a few swelling cells.The ischemic necrosis area of the spinal cord tissue was significantly reduced.The expression of NF-κB was decreased while that of IL-10 increased markedly.Compared with the IR group, the poly IC group showed a significant increase in the expression of IFNβ (117.23±6.06 vs 55.65±4.02, P<0.01) and a remarkably decrease in the expressions of TNFα (190.45±4.16 vs 201.82±2.18, P<0.01) and IL-1β (39.27±2.48 vs 50.59±1.47, P<0.01) at 48 hours.Conclusion Poly IC can protect the spinal cord and reduce inflammatory response after spinal cord ischemia-reperfusion injury.

Objective To investigate the inhibition of Eriobotryae Folium from twenty different districts towards phosphodiesterase 4(PDF4) in vitro.Methods The Eriobotryae Folium were extracted with 95％ ethanol reflux and the inhibition rates against PDE4D2 were carried out by liquid scintillation counting method.Results All the samples exhibited inhibitory activities towards PDE4 at 5 mg/L.Among them,nine samples were of the inhibition rate less than 80％,eleven samples were of more than 80％ inhibition and eight samples were of more than 90％ inhibition.Conclusion The Eriobotryae Folium shows significantly different inhibitory activities towards PED4.

Objective:To construct the expression vector of the fusion protein of human serum albumin (HSA) and thymopentin (TP5) and to express it in Pichia pastoris,and to elucidate the biological activity of fusion protein.Methods:The HSA-TP5 fusion gene was constructed by gene recombination and transfected into Pichia pastoris to construct the eukaryotic expression system of HSA-TP5.The recombinant eukaryotic expression plasmid of PPICZα-HSA-TP5 was obtained by agarose gel electrophoresis and purification reagent.The two step fermentation method was used to ferment gene engineering bacteria of HSA-TP5 in high density,and the fermentation supernatant protein was precipitated and concentrated;the purified fusion protein was obtained by cation exchange chromatography and hydrophobic chromatography and analyzed by SDS-polyacrylamide gel electrophoresis.The effect of the fusion protein on the proliferation of lymphocytes was detected by MTT assay.Results:The HSA target gene fragment with length of 1 845 bp was achieved by PCR method.The HSA-TP5-pPICZαC fusion plasmid was identified by restriction endonuclease digestion,and the fragment length was 707 bp.The sequence analysis showed that the HSA and TP5 sequences of the target genes were identical with the gene sequences reported in GenBank and were fused by forward fusion.PCR method confirmed that the eukaryotic recombinant plasmid PPICZ αC-HSA-TP5 was integrated into the yeast genome,and compared with control group,the target gene PCR product length was found to be 1 860 bp.SDS-PAGE analysis showed that the expression level of HSA-TP5 fusion protein was gradually increased with the induction time within 72 h.HSA-TP5 fusion protein was purified by cation exchange chromatography and AKTA multifunctional protein purification system.The MTT assay results showed that HSA-TP5 fusion protein was consistent with TP5 protein in promoting lymphocyte proliferation activity.Conclusion:HSA-TP5 fusion protein can be obtained by constructing the eukaryotic expression system of Pichia pastoris and owns the biological activity.

OBJECTIVETo downregulate the expression of pituitary tumor transforming gene 1 (PTTG1) in osteosarcoma (OS) cells by siRNA technology and to investigate related biological impact on cell proliferation, cell cycle and cell invasion of OS.

METHODSThree OS cell lines and osteoblast hFOB1.19 cell line were used in this study. Control siRNA and PTTG1 siRNA were employed to transfect OS U2OS cells, and PTTG1 protein level was detected by Western blot after the transfection. Effects of PTTG1 siRNA on cell proliferation, cell cycle and cell invasion were investigated by CCK-8, flow cytometry and Boyden chamber, respectively. Finally, activity of Akt and its downstream target gene expression were analyzed by Western blot in U2OS cells upon various treatments.

RESULTSExpression of PTTG1 protein in 3 OS cells (MG-63, SaOS-2 and U2OS) was significantly higher than that in osteoblast hFOB1.19, among which U2OS cells displayed the highest level. PTTG1 siRNA markedly downregulated the expression of PTTG1 protein in U2OS cells, leading to obvious inhibition of cell proliferation, altered cell cycle distribution and reduced ability of invasion of U2OS cells. Moreover, downregulation of PTTG1 reduced the expression of p-Akt (S473 and T308), MMP-2 and MMP-9 proteins, along with enhanced expression of p21 and E-cadherin proteins.

CONCLUSIONSPTTG1 may be tightly linked to the development of OS and therefore may serve as a novel target for precision therapy of OS.

Bone Neoplasms ; metabolism ; pathology ; Cadherins ; metabolism ; Cell Cycle ; drug effects ; physiology ; Cell Movement ; Cell Proliferation ; drug effects ; physiology ; Down-Regulation ; Humans ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Neoplasm Invasiveness ; Osteosarcoma ; metabolism ; pathology ; RNA, Small Interfering ; pharmacology ; Securin ; genetics ; metabolism ; Transfection

Objective To investigate the efficacy and safety of cool-tipped saline-irrigated catheter ablation of epicardial accesso-ry pathway in the patients after failure of conventional radiofrequency ablation .Methods 30 patients with epicardial accessory path-way after failuer of conventional radiofrequency ablation were included in this study ,among them ,25 cases of the left accessory pathway and 5 cases of the right accessory prathway used the cool-tipped catheter for conducting ablation through the endocardium (ventricular side or atrial side) or coronary sinus .Results All patients achieved success ,no intraoperative and postoperative related complications occurred .Conclusion The cool-tipped saline-irrigated catheter ablation of epicardial accessory pathway is effective and safe in the patients after failure of conventional radiofrequency ablation ,which may improve the success rate of epicardial acces-sory pathway ablation .

OBJECTIVETo identify a HEK293 cell line containing stably-transfected H3R gene, and to screen the novel non-imidazole compounds with H3R antagonist activity.

METHODSThe expression of rat H3 receptor in cell line was detected by RT-PCR and Western blot. An elevation of intercellular cAMP concentration induced by forskolin was measured as the index for screening compounds with H3R antagonist activity.

RESULTSThe H3R-transfected HEK-293 cells stably expressed high level of rat H3 receptor mRNA and protein. Forskolin significantly increased intercellular cAMP concentration in the H3R-transfected HEK-293 cells. H3R agonist (R)-α-methylhistamine inhibited the forskolin-induced production of intercellular cAMP. H3R antagonist thioperamide and newly synthesized non-imidazole compounds XHA23 and XHA25 blocked (R)-α- methylhistamine reversal of forskolin-induced cAMP formation in a concentration-dependent manner, and the IC50 values were 3.62 μmol/L, 0.49 μmol/L, 0.14 μmol/L, respectively.

CONCLUSIONThe H3R-transfected HEK293 cells stably express high level of rat H3 receptor, and can be used for screening compounds with H3R antagonist activity. The non-imidazole compounds XHA23 and XHA25 may have H3R antagonist activity.

Animals ; Drug Evaluation, Preclinical ; HEK293 Cells ; Histamine H3 Antagonists ; Humans ; Rats ; Receptors, Histamine H3 ; genetics ; metabolism ; Transfection

OBJECTIVE: To evaluate the therapeutic results of endoscopic orbital decompression for thyroid-associated ophthalmopathy. METHOD: The records of nine patients (twelve orbits) received endoscopic orbital decompression for thyroid-associated ophthalmopathy were analyzed for changes in visual acuity, intraocular pressure, proptosis, corneal ulceration and movement. The follow-ups ranged from two months to thirty-six months. RESULT: Twelve orbits (100%) had improvement in visual acuity (range 0.1-0.7). Ten orbits (83.3%) decreased in intraocular pressure (range 0.2-21.4 mm Hg). Eight orbits (66.70%) decreased in proptosis (one-five mm). The orbit with corneal ulcer was healed after decompression. Diplopia was cured in one of four patients. CONCLUSION Endoscopic orbital decompression is a safe and effective procedure for the treatment of thyroid-associated ophthalmopathy.
Adult ; Decompression, Surgical ; methods ; Endoscopy ; Female ; Graves Ophthalmopathy ; surgery ; Humans ; Male ; Middle Aged ; Orbit ; surgery ; Retrospective Studies ; Treatment Outcome

Objective To investigate the relationship of external hydrocephl(EH) with brain-injure in the neonate.Methods By performing on the patients with brain-injure with CT and making the clinical study,the change of brain following injure and its relationship with EH were analyzed.Results Hypoxic-ischemic encephalopathy(HIE) were 36 cases,EH were 19 in the cases,6 get the cerebrospinal symptom;SAH were 22 cases,EH were 15 in the cases,3 get the cerebrospinal symptom;Intracranial hemorrhage were 8 cases,EH were 3 in the cases,3 got the cerebrospinal symptom.Conclusion All the kinds of brain-injure can get EH and the rate of EH with any brain-injure is no different.But the outcome is significantly different.