Objective:To investigate the effect of melatonin on the expression of PD-L1 on the surface of ovarian cancer cells.Methods:Ovarian cancer cell line OVCAR3 was treated with melatonin,then flow cytometry was used to detect the expression level of PD-L1 on the surface of ovarian cancer cells.Western blot was used to detect the expressions of PD-L1 and LC-3 in ovarian cancer cells after different treatments.After adding autophagy inhibitor Autophinib,flow cytometry was used to detect the expression of PD-L1 on the surface of ovarian cancer cells,ovarian cancer cells were treated with melatonin or melatonin combined with autophagy inhibi-tors and co-incubated with human T lymphocyte Jurkat.The proportion of ovarian cancer cell apoptosis was detected by flow cytometry.Results:Melatonin treatment significantly reduced expression of PD-L1 on the surface of ovarian cancer cells,promoted autophagy of ovar-ian cancer cells.Autophagy inhibitors reversed down regulation of PD-L1 treated by melatonin,Jurkat cells killed more melatonin treated ovarian cancer cells,and the killing of ovarian cancer cells by Jurkat cells revised by autophagy inhibitors.Conclusion:Mela-tonin can enhance the killing effect of T cells on ovarian cancer cells.

Neck dissection(ND)is one of the main treatment methods for oral and maxillofacial malignancies.Although ND type is in con-stant improvement,but intraoperative peal-pull-push injury of the accessory nerve,muscle,muscle membrane,fascia and ligament induced shoulder syndrome(SS)is still a common postoperative complication,combined with the influence of radiochemotherapy,not only can cause pain,stiffness,numbness,limited dysfunction of shoulder neck and arm,but also may have serious impact on patient's life quality and phys-ical and mental health.At present,there is still a lack of a systematic evaluation and rehabilitation management program for postoperative SS of oral and maxillofacial malignant tumors.Based on the previous clinical practice and the current available evidence,refer to the relevant lit-erature at home and abroad,the experts in the field of maxillofacial tumor surgery and rehabilitation were invited to discuss,modify and reach a consenusus on the etiology,assessment diagnosis,differential diagnosis,rehabilitation strategy and prevention of SS,in order to provide clinical reference.

Objective:To investigate the diagnostic efficacy of miR-9 and miR-195-3p for primary hepatic carcinoma (PHC), and the changes in miR-9 and miR-195-3p levels after interventional therapy.Methods:A total of 123 cases of PHC patients and 30 cases of liver cirrhosis patients attending Tangshan People's Hospital from May 2019 to May 2020 were selected as the PHC group and the liver cirrhosis group, respectively, and 50 people who were physically healthy during the same period were selected as the healthy group. Serum miR-9 and miR-195-3p levels were detected by real-time quantitative PCR. The relationship between serum miR-9 and miR-195-3p levels and clinical-pathological characteristics of PHC patients was analyzed. Receiver operator characteristic (ROC) curve was applied to analyze the diagnostic efficacy of miR-9 and miR-195-3p for PHC. The changes in serum miR-9 and miR-195-3p levels in PHC patients before and after transcatheter arterial chemoembolization (TACE) were compared.Results:There were statistically significant differences in serum miR-9 (0.99±0.10, 1.31±0.28, 1.68±0.43) and miR-195-3p (0.97±0.10, 0.83±0.22, 0.63±0.18) levels among the healthy group, liver cirrhosis group, and PHC group ( F=69.78, P<0.001; F=74.82, P<0.001), with serum miR-9 levels increased successively and miR-195-3p levels decreased successively among the three groups (all P<0.05). There were statistically significant differences in serum miR-9 ( t=7.45, P<0.001; t=5.32, P<0.001; t=4.96, P<0.001) and miR-195-3p ( t=16.17, P<0.001; t=4.21, P<0.001; t=7.53, P<0.001) levels in PHC patients with different maximum diameters of tumor, clinical stages and degrees of differentiation. ROC curve analysis showed that the area under the curve (AUC) for the combined differential diagnosis of liver cirrhosis and PHC by serum miR-9 and miR-195-3p testing was 0.919, which was higher than the AUC for the differential diagnosis of serum miR-9 (AUC: 0.712, Z=4.38, P<0.001) and miR-195-3p (AUC: 0.844, Z=2.04, P=0.042) alone. After TACE treatment, serum miR-9 levels decreased (1.39±0.21 vs. 1.68±0.43, t=14.22, P<0.001) and miR-195-3p levels increased (0.78±0.22 vs. 0.63±0.18, t=14.84, P<0.001) in patients compared to pre-treatment levels. Conclusion:Serum miR-9 level is increased and miR-195-3p level is decreased in patients with PHC compared with patients with liver cirrhosis and healthy subjects, and the combination of the two has high differential diagnostic efficacy for liver cirrhosis and PHC. After TACE treatment, serum miR-9 level is decreased and miR-195-3p level is increased in PHC patients.

ObjectiveTo investigate the effect of hypoxia-inducible factor-1α (HIF-1α) on the stemness and epirubicin sensitivity of hepatoma cells. MethodsHepatoma cells were selected for experiment. HepG2 hepatoma cells transfected with HIF-1α overexpression plasmid were selected as experimental group, and those transfected with pcDNA3.1 empty plasmid were selected as control group; HepG2 cells alone were selected as HepG2 group. Quantitative real-time PCR was used to measure the mRNA expression of HIF-1α; Western blot was used to measure the protein expression of HIF-1α; flow cytometry was used to measure the expression of CD133 on the surface of hepatoma cells. The three groups of cells were treated with epirubicin at different concentrations (0, 6.25, 12.5, 25, and 50 μmol/L) for 24 hours; MTT assay was used to measure cell viability, and flow cytometry was used to measure apoptosis after treatment with epirubicin (50 μmol/L). A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the t-test was used for further comparison between two groups. ResultsCompared with the HepG2 group and the control group, the experimental group had a significant increase in the mRNA expression of HIF-1α (both P＜0.001), and Western blot showed high expression of HIF-1α in the experimental group. The percentage of CD133 cells was 0.040%±0.003% in the HepG2 group, 0.030%±0.010% in the control group, and 20.110%±0.600% in the experimental group, and the experimental group had a significantly higher positive rate of CD133+ than the HepG2 group and the control group (both P＜0.001). At an epirubicin concentration of 25 and 50 μmol/L, the HepG2 group and the control group had significantly inhibited cell viability and a significantly lower cell viability than the experimental group (both P＜005). After the treatment with 50 μmol/L epirubicin for 48 hours, the experimental group had a significantly lower cell apoptosis rate than the HepG2 group (67.9%±2.5% vs 93.6%±1.5%, P＜0.001) and the control group (67.9%±2.5% vs 93.0%±1.2%, P＜0001). ConclusionHepG2 cells are successfully transfected with HIF-1α overexpression plasmid, and HIF-1α can increase the percentage of liver cancer stem cells and improve their resistance to epirubicin.

Objective:To analyze the expression and clinical significance of lymphoid enhancer factor-1 (LEF-1) and P53 protein in hilar cholangiocarcinoma.Methods:A total of 50 cases with hilarcholangiocarcinoma in the Tangshan People′s Hospital from March 2010 to December 2017 were retrospectively analyzed.P53 protein mutation was detected by immunohistochemistry.At the same time, the expression of LEF1 protein in cholangiocarcinoma and adjacent normal tissues was detected.The expression of LEF1 and its correlation with clinicopathological parameters were analyzed.At the same time, the relationship between p53 protein mutation and LEF1 protein expression and the prognosis of patients were discussed.Results:The positive expression rate of LEF1 in tumor tissues were 62.00%(31/50), which was higher than in adjacent tissues 36.00% (18/50). The difference was statistically significant (χ 2=6.763, P=0.016). There were 28 patients with TP53 protein mutation and 22 patients with wild type.The positive rate of LEF1 in TP53 mutant group was 75.00% (21/28), which was significantly higher than that in TP53 wild type group(45.45%, 10/22), and the difference was statistically significant(χ 2=4.565, P=0.03). The LEF1 expression was associated with tumor differentiation, TNM stage, lymph node metastasis and TP53 protein mutation (all P<0.05). LEF-1 expression was positively correlated with TP53 protein mutation ( r=0.294, P=0.04). The 1-year cumulative survival rate of patients with TP53 protein mutation was significantly lower than that of patients with TP53 protein wild type (60% vs.81%, P=0.0416). The 1-year cumulative survival rate of patients with LEF1 positive expression was also significantly lower than that of patients with negative expression (53% vs.82%, P=0.0180). Conclusion:LEF-1 is highly expressed in hilar cholangiocarcinoma, and patients with high expression have poor prognosis.The positive expression of LEF-1 in hilar cholangiocarcinoma patients with TP53 protein mutation was increased, and the prognosis of patients with TP53 protein mutation was poor.

Salvianolic acid A (SalA) is an effective compound extracted from traditional Chinese medicine Bunge. The Forkhead box O3a (FOXO3a) signaling pathway plays crucial roles in the modulation of ischemia-induced cell apoptosis. However, no information about the regulatory effect of SalA on FoxO3a is available. To explore the anti-cerebral ischemia effect and clarify the therapeutic mechanism of SalA, SH-SY5Y cells and Sprague-Dawley rats were applied, which were exposed to oxygen glucose deprivation/reoxygenation (OGD/R) and middle cerebral artery occlusion/reperfusion (MCAO/R) injuries, respectively. The involved pathway was identified using the specific inhibitor LY294002. Results showed that SalA concentration-dependently inhibited OGD/R injury triggered cell viability loss. SalA reduced cerebral infarction, lowered brain edema, improved neurological function, and inhibited neuron apoptosis in MCAO/R rats, which were attenuated by the treatment of phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K) specific inhibitor LY294002. SalA time- and concentration-dependently upregulated the phosphorylation levels of protein kinase B (AKT) and its downstream protein FOXO3a. Moreover, the nuclear translocation of FOXO3a was inhibited by SalA both and , which was also reversed by LY294002. The above results indicated that SalA fought against ischemia/reperfusion damage at least partially the AKT/FOXO3a/BIM pathway.

Objective To compare the short term clinical efficacy of percutaneous resection and laparoscopic deroofing of renal cysts.Methods From October 2013 to June 2016,the data from 39 cases with renal cysts were followed for approximately 22 months (ranging 5-36 months).Patients were randomized into two groups.In the resectoscopic group(17 pts),the mean age of those patients was 57 years (ranging 34-81 years).The mean size of these cysts was 6.4cm (ranging 5.4-8.2 cm).The mean preoperative creatinine was 66.5μmol/L (ranging 38.1-108.8μ mol/L).The mean preoperative sodium was 141.4μ mol/L(ranging 135.6-145.1μmol/L).The mean preoperative potassium was 4.0μmol/L (ranging 3.4-4.7 μmol/L).In the latter laparoscope (22pts),the mean age of these patients was 60 years (ranging 46-73 years).The mean size of these cysts was 6.8cm (ranging 4.3-8.9cm).The mean preoperative creatinine was 74.8μmol/L (ranging 48.6-141.9μmol/L).The mean preoperative sodium was 141.5μmol/L(ranging 135.0-146.1 μmol/L).The mean preoperative potassium was 4.0μmol/L (ranging 3.1-4.8μmol/L).The operative time,blood loss,days of drainage,hospital stay and complications were compared with the two groups.Results All of the 39 cases were accepted the procedure successfully without open conversion.Compared the resectoscopic group with Laparoscopic,the mean operative time was 29.7 min (ranging 15-50 min) and 63.0min (ranging 20-1 00 min),mean blood loss was 36.6ml(ranging 10-80 ml) and 60.4ml(ranging 10-200 ml),days of drainage was 2.3 days and 3.1 days,hospital stay was 3.7 days and 5.1 days,the mean postoperative creatinine was 67.4 μmol/L(ranging 43.8-95.5 μmol/L) and 68.9μmol/L(ranging 46.5-157.6 μmol/L),the mean postoperative sodium was 141.2μmol/L(ranging 136.0-147.2 μmol/L) and 141.6 μmol/L(ranging 136.0-147.2 μmol/L),the mean postoperative potassium was 3.8 μmol/L (ranging 3.2-4.3 μmol/L) and 3.8μmol/L (ranging 3.3-4.3 μmol/L).The overall postoperative pathology was renal cysts.All cases were followed for approximately 19 months (ranging 6-35 months) and 22 months (ranging 5-36 months) in reseetoscopic and laparoscopic group respectively.No cysts recurrence was found by ultrasound.Conclusions Compared with laparoscopic deroofing,use of resectoscopic technology significantly enhances the possibility of achieving better intraoperative results in all patients with renal cysts.Percutaneous resection of simple renal cysts is safe and feasible.

Objective To study polymorphism of the free radical scavenger edaravone and to find the one which has good advantages to the clinical medication. Methods Preparation of four forms of edaravone through kinds of physical or chemical methods. These polymorphs were characterized by single crystal X-ray diffraction method (SXRD), powder X-ray diffraction method (PXRD), differential scanning calorimetry method (DSC), infrared spectrum method (IR) and melting point method (MP);a variety of influence factors experiment were used to research the stability of polymorphs. Solid edaravone in different forms were orally administered to SD rats respectively, the plasma concentration of the drug was determined by HPLC, and the pharmacokinetic characteristic of different forms was compared according to the HPLC results. Results Four forms(form A, form B, form C, form D) and the preparation of pure crystal forms were obtained by crystal screening technology. These polymorphs were identified by PXRD, DSC and IR. The edaravone polymorphs have an influence on the stability and pharmacokinetic. Conclusion Edaravone research provideds a variety of crystal material composition, preparation methods, stability, solubility and pharmacokinetic characteristic, and form A has been proved of good advantage one. This research provides data and technology support for choice of preponderant pharmaceutical polymorphs and drug quality standard improvement.

OBJECTIVE To evaluate the effect of single traditional Chinese medicine(TCM) herb extracts on hepatoma and normal fibroblast cells using high-throughput screening in order to obtain extracts with specific anti-hepatoma effect. METHODS 242 commonly used TCM herbs were extracted by petroleum ether,ethanol and water,respectively. The total number of TCM extracts was 554. The cyto?toxicity of samples was evaluated by MTT in human hepatoma cells Bel7402 and mice normal fibroblasts NIH3T3. RESULTS 7.4%of the total extracts had an inhibitory effect greater than 50%for Bel7402,but 14.8% for fibroblasts NIH3T3 cells. Extracts with an inhibitory effect above 50% on both Bel7402 and NIH3T3 cells accounted for 4.4%of the total extracts. Our results showed that the sample DF173 had preferable cytotoxicity effect on hepatoma carcinoma cells in a good dose-effect relationship. DF173 is an ethanol extract from Stephania tetrandra,which is a commonly used herb in TCM. The cytotoxic IC50 of DF173 against Bel7402 was 8.27 mg·L-1,and 19.48 mg·L-1 on NIH3T3. CONCLUSION The components of TCM herbs are highly complicated. The combination of tumor cells with normal fibroblast cells to evaluate the cytotoxicity effect during anti-tumor drug screening will contribute much to the discovery of TCM drugs with high anti-tumor efficiency and lower toxicity.

Objective The decease of varicose vein and telangiectasis of lower extremity has always been difficult to treat.Regular treatments and medicines are invalid.This research aimed to study the effect of foam sclerotherapy and the long-pulsed 1064 nm Nd∶YAG laser in treatment of reticular veins and telangiectasis of lower extremity.Methods From January 2012 to June 2012,excluding deep and superficial veins valve insufficiency of the lower extremity through duplex uhrasonography,72 patients with simple reticular veins and telangiectasis of the lower extremity were treated with foam sclerosing agent injection combined with Nd∶YAG 1064 nm laser therapy.Results Of the 72 patients,cure was achieved in 38 cases,significantly effective in 32 cases,effective in 2 cases,and total effective rate was 100％.Follow-up showed that after 3 months 1 case was dissatisfied,3 cases were discontent,6 relatively satisfied,32 satisfied,and 30 very satisfied;after 12 months no cases were very dissatisfied,1 cases were discontent,9 relatively satisfied,28 satisfied,and 34 cases very satisfied.There were no severe complications in all cases.Conclusions Foam sclerotherapy and Nd:YAG 1064 nm laser can be used for different stages of the treatment process and different caliber of blood vessels.Treatment with foam sclerotherapy and Nd∶YAG 1064 nm laser for reticular veins and telangiectasia of lower extremity is a safe,simple and effective way.