Objective To investigate the influence of isoquercitrin on the inflammatory factors in LPS-induced RAW264.7 cells.Methods MTT method was used to detect inhibition ratio of RAW264.7 cells induced by isoquercitrin.The level of TNF-α in culture medium was measured by ELISA.Nitric oxide (NO) was detected by Nitrate Assay Kit.Western blotting was used to investigate the influence on the productions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2).Results The half inhibitory concentration (IC50) of isoquercitrin was 65.73 μmol·L-1.LPS had no inhibitory effect on the cells.Compared with LPS group,the level of TNF-α was decreased to 74.80% and 60.57% in isoquercitrin (20,10 μmol·L-1) groups in a dose-dependent manner.The results measured by Nitrate Assay Kit revealed that isoquercitrin (20,10 μmol·L-1) could suppress production of NO,the level of NO decreased to 79.34% and 68.81%(P<0.05).The Western blotting results showed that isoquercitrin (20,15,10 μmol·L-1) inhibited the productions of iNOS and COX-2 (P<0.05).Conclusion Isoquercitrin has anti-inflammatory effects by inhibiting the productions of TNF-α,NO,iNOS and COX-2,and the most effective dose for the inhibition is 10 μmol·L-1.

Aim To study the inhibitory effect of isoquercitrin on Raf/MEK/ERK signaling pathway in HepG2 cells.Methods MTT was used to detect the proliferation of human liver cancer HepG2 cells after the treatment of isoquercitrin.The morphology and growth of cells were observed under inverted microscope after the different concentrations of isoquercitrin(0, 40, 80, 160, 320 μmol·L-1) to treat HepG2 cells for 24 and 48 h.Cell cycle was assessed by flow cytometry.Ras, Raf, MEK, ERK expression was assayed by Western blot, and mRNA expression was detected by quantitative fluorescence PCR.Results Isoquercitrin could inhibit the growth of HepG2 cells in a concentration-and time-dependent manner.Typical morphological changes of apoptosis were observed by inverted microscopy after HepG2 cells were treated with different concentrations of of isoquercitrin for 24 h or 48 h.The cell cycle assay showed that with the increasing concentration of isoquerditrin, the number of cells that was arrested in G1 phase gradually increased.Compared with the blank group, the expressions of Ras, Raf, MEK, ERK mRNA were down-regulated, and related proteins expression were also down-regulated(P<0.05), and these results had statistical significance.Conclusion Isoquercitrin can induce the apoptosis of HepG2 cells, which may be related to the Raf/MEK/ERK signaling pathway.

Objective To evaluate the therapeutic effect of combination of sandostatin and growth hormone (GH) in the treatment of severe acute pancreatitis (SAP ). Methods Sixty patients with SAP were divided randomly into 3 groups:(1)Sandostatin treatment(ST) group (n=15);(2)combination of sandostatin with GH treatment(CT) group (n=30) ;(3)control group (n=15). The changes in serum IL-1, IL-6,TNF-? and albumin levels after treatment, and the incidence of complications, the duration of hospital stay and cost were compared among the 3 groups. Results The complications, mortality, duration of hospital stay in the CT group were significantly shorter than those in ST group and control group (all P

Objective To investigate whetherthe extract of HUANGPI inhibitthe secretion of TNF-αvia TLR4/MyD88/TRAF6 signaling pathway. Methods ELISA assay was performed to determine TNF-α level in cell culture medium. MTT assay was used to detect the effects of the extract of HUANGPI and LPS on the viabilities of RAW 264.7 cells. Proteinexpressions of TLR4 and TRAF6 were detected by Western blotting assay. Results The extract of HUANGPI inhibited the secretion of TNF-αin a dose-dependent manner. Compared to LPS group , were TNF-αwas significantly suppressed in the cells in LPS+MyD88 inhibitor group , LPS+extract group and LPS+extract+MyD88 inhibitor group,with the corresponding reductions of TLR4 and TRAF6 protein expression at74% and21%,70% and27%,44% and8.5%, respectively. Conclusion MYD88-dependent signaling pathway might be involved in the mechanism underlying the effect of the extract of HUANGPI on suppressing LPS-induced inflammation.

Objective To explore the effect of proprioceptive neuromuscular facilitation (PNF) rehabilitation training on functional recovery of athletic rotator cuff injury. Methods Twenty-two college students with athletic rotator cuff injuries were stratified according to their gender and randomly divided into resistance band + passive joint range of motion (ROM) training group (control group) and PNF training group (experimental group). The visual analog scale (VAS) was used to evaluate subjective pain intensity of the shoulder joint. Pain positive rate for each manipulation test of rotator cuff injury was observed, and active ROM and muscle strength of the shoulder joint were measured. Improved UCLA shoulder joint score was used to evaluate comprehensive function of the shoulder joint. Results After training, VAS scores and pain positive rate in two groups were lower than those before training, and VAS scores and pain positive rate in experimental group were lower than those in control group. Muscle strength, active ROM in all directions and improved UCLA score of the shoulder joint in two groups were also higher than those before training, and the internal rotation muscle strength, the internal rotation and external rotation active ROM, improved UCLA score of the shoulder joint in experimental group were higher than those in control group. Conclusions PNF rehabilitation training can reduce the pain of athletic rotator cuff injury, improve the active ROM, muscle strength and UCLA shoulder joint score. The function recovery effect of PNF training is better than that of resistance band + passive ROM training.

Objective: To investigate the effects of long non-coding RNA nuclear enriched abundant transcript 1 (lncRNA NEAT1) on the proliferation of lung adenocarcinoma PC-9 cells and to explore its mechanism. Methods: qPCR was used to detect the expression level of lncRNA NEAT1 in human lung adenocarcinoma PC-9 cells and human embryonic lung diploid 2BS cells. The sequence of small interfering RNA(siRNA) targeting lncRNANEAT1 gene was designed and synthesized, and then transfected into PC-9 cells by liposome method. The expression level of NEAT1 in PC-9 cells before and after transfection was detected by qPCR. MTT and flow cytometry were used to detect the effect of lncRNANEAT1 knockdown on proliferation and cell cycle distribution of PC-9 cells, respectively. WB assay was used to detect the expressions of DNA damage-related proteins, namely, double-stranded DNA breaks (DSBs) biomarker γ-H2AX and ataxia-telangiectasia mutated (ATM), before and after transfection. Results: Compared with 2BS cells, lncRNA NEAT1 was highly expressed in PC-9 cells (P<0.05). The PC-9 cells with lncRNA NEAT1 knock-down were successfully established. After being transfected with siRNA for 12 h, the proliferation of PC-9 cells in siNEAT1 group and siNEAT2 group significantly decreased as compared with the blank control group and the empty transfection group (P<0.05). In the interference groups, cell cycle was arrested in G1 phase ([88.97±2.64]%, [88.15±1.48]% vs [84.5±1.72]%, P<0.05) and G2/M phase ([8.35±2.02]%, [8.11± 1.36]% vs [4.28±1.28]%, P<0.05). The expression levels of DNA damage-related proteinsATM and γ-H2AX in the interference groups were significantly increased (all P<0.05). Conclusion: lncRNA NEAT1 is highly expressed in lung adenocarcinoma PC-9 cells. lncRNA NEAT1 inhibits DNA damage and causes cell cycle at G1/M phase switch, and thus promotes the proliferation of lung adenocarcinoma cells.

Objective: To study the miR-28-3p expression in triple negative breast cancer (TNBC) tissues and cell lines, and explore its effect on the malignant biological behaviors of MDA-MB-468 cells. Methods： ：Tumor tissues and matched para-cancerous tissues were collected from 83 TNBC patients, who underwent tumor resection and pathological confirmation in the Fourth Hospital of Hebei Medical University between Jan. 2013 and Jan. 2014. TNBC cell lines (MDA-MB-468, HCC-1937, MDA-MB-231, MDA-MB436, MDA-MB-453) and human normal breast epithelial cell line MCF10A were also used in this study. qPCR was used to detect the expression of miR-28-3p in above mentioned tissues and cell lines. The correlation between miR-28-3p expression and clinical parameters was analyzed.After transfection with miR-28-3p inhibitor, the proliferation, apoptosis, invasion and migration ability of MDA-MB468 cells were detected with CCK-8, Flow cytometry, Transwell and Wound-healing experiment, respectively. And Western blotting was used to examine the protein expression of bridging integrator-1 (BIN1) in MDA-MB-468 cells. Bioinformatics BIN1 tool waere used to predict the target gene of miR-28-3p. Luciferase reporter gene assay was performed to validate the regulatory effect of miR-28-3p on BIN1. Results: The expression of miR-28-3p in TNBC tissues and cell lines was higher than that in matched paracancerous tissues and MCF10Acells (all P<0.01), respectively.Among the total 83 TNBC tissues, 56 (67.47%) showed high miR-28-3p expression. High expressionofmiR-28-3pwascloselycorrelated with the Ki-67 expression, tumor size and TNM stage (all P<0.05 or P<0.01). Compared with miR-NC group, transfection of miR-28-3p inhibitor significantly decreased the proliferation, invasion and migration of MDA-MB-468 cells while increased the apoptosis rate (all P<0.05 or P<0.01). Luciferase reporter gene assay confirmed that BIN1 was a target gene of miR-28-3p, and miR-28-3p inhibitor could up-regulate BIN1 expression in MDA-MB-468 cells (P<0.05). Conclusion: miR-28-3p is highly expressed in TNBC tissues and cell lines. miR-28-3p inhibitor up-regulates the expression of BIN1 to inhibit the proliferation, invasion and migration ability while promote the apoptosis of MDA-MB-468 cells.

[Abstract] Objective：To study the expression of long non-coding RNA(lncRNA) titin antisense RNA1 (TTN-AS1) in lung adenocarcinoma (LUAD) tissues, and explore its relationship with clinicopathologic characteristics and prognosis of LUAD patients. Methods: The TTN-AS1 expression in LUAD data set was analyzed using TCGAdatabase. 52 pairs of tumor tissues and matched para-carcinoma tissues from LUAD patients, who underwent surgical resection and were later pathologically conformed in Fourth Hospital of Hebei Medical University between Jan. 2014 and Jan. 2015, were used in this study. qPCR was performed to detect TTN-AS1 expression in the specimens. Then, the correlations between TTN-AS1 expression and clinicopathologic characteristics were analyzed. Survival analysis was used to determine the significance of TTN-AS1 expression for predicting the prognosis of LUAD patients. Results: TCGAdatabase analysis and qPCR results showed that TTN-AS1 expression in LUAD tissues was significantly higher than that in normal lung and para-carcinoma tissues (both P<0.01). TTN-AS1 expression in LUAD tissues was significantly correlated with the TNM stage and lymph node metastasis (P<0.05), but not correlated with gender, age, tumor invasion range (P>0.05). Kaplan-Meier univariate analysis result demonstrated that the patients with high TTN-AS1 expression had shorter post-operative disease-free survival (DFS) and overall survival (OS) than those patients with low TTN-AS1 expression (all P<0.01). Cox proportional hazard regression model result demonstrated that wider tumor invasion range, positive lymph node metastasis and high TTN-AS1 expression were significantly correlated with shorter postoperative DFS and OS (P<0.05). Conclusion: TTN-AS1 was highly expressed in LUAD tissues, and closely correlated with TNM stage and lymph node metastasis of LUAD patients (all P<0.05). High expression of TTN-AS1 is significantly correlated with shorter DFS and OS, indicating that TTN-AS1 may be a biomarker for predicting poor prognosis of LUAD patients.

[摘 要] 目的：探讨PD-1抗体联合化疗对比抗血管生成药物联合化疗在晚期驱动基因阴性肺腺癌一线治疗中的疗效和安全性。方法：收集2018年3月至2021年8月河北医科大学第四医院收治的141例不可手术切除的ⅢB/ⅢC和Ⅳ期驱动基因阴性肺腺癌患者，回顾性分析PD-1抗体联合化疗对比抗血管生成药物联合化疗在一线治疗中的疗效与安全性。主要研究终点为无进展生存期（PFS），次要终点为客观缓解率（ORR）、疾病控制率（DCR）和不良反应。结果：141例患者均纳入生存分析，中位随访时间为13.0个月（95% CI：12.0～14.0）。PD-1抗体联合化疗组（A组）和抗血管生成药物联合化疗组（B组）的ORR分别为33.33%和27.38%，DCR分别为98.25%和89.29%，差异均无统计学意义。A组和B组的中位PFS分别为8.4个月（95% CI: 7.3～9.9）和6.9个月（95% CI: 6.1～7.7），差异无统计学意义。亚组分析结果显示，ⅢB/ⅢC期、肝或脑转移患者中，A组中位PFS较B组均延长（均P<0.01）。A组和B组不良反应发生率分别为26.32%和14.29%，多数为1~2级。结论：PD-1抗体联合化疗对比抗血管生成药物联合化疗一线治疗晚期驱动基因阴性肺腺癌疗效相当，不良反应可耐受，可成为晚期驱动基因阴性肺腺癌标准一线治疗。