The non-structural protein 5(NS5)is a highly conserved protein in the Flavivirus genus,acting as both a methyltransferase(MTase)and an RNA-dependent RNA polymerase(RdRp).It has been well documented that NS5 plays a crucial role in the replication of viral RNA.Recent studies have shown that NS5 proteins from different flaviviruses interact with various proteins in host cells,aiding the virus in evading the immune system.This review summarizes the structure,subcellular localization,and function of NS5 proteins.Additionally,we outline how flavivirus NS5 proteins contribute to viral replication and immune evasion.Lastly,we present the recent developments of specific small molecule inhibitors that target NS5 proteins.

Objective:To compare the postoperative anorectal and urogenital function in patients undergoing Da Vinci robot vs laparoscopic total mesorectal excision (TME) for middle and low rectal cancer.Methods:A prospective controlled study was conducted to analyze the clinical data of 120 patients with middle and low rectal cancer receiving low anterior resection (Dixon procedure) at the Department of Gastrointestinal Surgery, Guizhou Provincial People's Hospital from Jun 2020 to Oct 2022, including 65 patients in the laparoscopic surgery group and 55 patients in the robotic surgery group.All patients underwent colonoscopy and pelvic MRI, and the distance of the tumor from the anal margin was less than 10 cm.The clinical data of the patients were collected and followed up by questionnaire at 12 months after operation. The anal defecation function was assessed by Wexner constipation score and low anterior resection syndrome scale (LARS).The urinary function was measured by International Prostate Symptom Score (IPSS) and International Advisory Committee on Urinary Incontinence Lower Urinary Tract Symptom Scale (ICIQ-MLUTS/ ICIQ-FluTS).Reproductive function was valued by International Index of Erectile Function (IIEF-5 score) was used for male function and the sexual function index (FIFS-19) for females.Results:The postoperative anal defecation function in robotic group was better than that of the laparoscopic group, and the LARS score was (4.3±2.2) vs. (9.8±1.5), t=9.151, P=0.038.There was no serious urinary dysfunction in neither groups. The robot group had a certain advantage in the protection of male urinary function [ICIQ-MLUTS, (1.8±5.8) vs. (13.8±4.9), t=4.128, P=0.038], while there was no significant difference in the female urinary function between the two groups .ICIQ-FLUTS [(-0.3±1.0) vs. (-0.2±0.9), t=0.015, P=0.844].There was no significant difference in reproductive function between the two groups, IIEF-5 score [(-13.4±2.7) vs. (-11.7±3.4), t=0.35, P=0.615]. FIFS-19 [(-5.2±4.6) vs. (-10.5±6.4), t=4.128, P=0.254]. Conclusions:Compared with laparoscopic surgery, robotic surgery has a better possibility of anal defecation after middle-low rectal cancer surgery. The robotic group has certain advantages in male urinary function protection, and the two surgical methods have similar effects on reproductive function protection.

OBJECTIVE To study the effects of the active component 17-hydroxy-jolkinolide B （HJB） of Euphorbia fischeriana on the proliferation and apoptosis of human triple-negative breast cancers （TNBC） MDA-MB-231 and MDA-MB-468 cells. METHODS MTT assay was adopted to detect the inhibitory rate of MDA-MB-231 and MDA-MB-468 cells proliferation after treated with 0 （blank control），5，10，20，40，80 μmol/L HJB for 24， 48 and 72 h. Laser confocal microscope and flow cytometry were adopted to detect the apoptosis， mitochondrial membrane potential（MMP） and reactive oxygen species （ROS） of above 2 kinds of cells after treated with 0 （blank control）， 10，20，40 μmol/L HJB for 24 h. Western blot assay was used to detect the expressions of B cell lymphoma-2（ Bcl-2）， Bcl-2-associated X protein （Bax）， cytochrome-C （Cyt-C）， caspase-3， cleaved caspase- 3， caspase-9 and cleaved caspase-9. RESULTS Compared with blank control group， 5，10，20，40，80 μmol/L HJB could significantly increase the inhibitory rate of MDA-MB-231 and MDA-MB-468 cells proliferation （P＜0.05）， in dose- and time- dependent trend. After 24 h treatment of HJB （10，20，40 μmol/L）， the apoptosis of above 2 kinds of cells increased， and the total apoptotic rate increased significantly （P＜0.05）； the mitochondrial membrane potential decreased significantly （P＜0.05）； the level of ROS increased significantly （P＜0.05）； the protein expressions of Bcl-2， caspase-3 and caspase-9 were decreased significantly （P＜ 0.05）， while the protein expressions of Cyt-C， Bax， cleaved caspase-3 and cleaved caspase-9 were increased significantly （P＜0.05）. CONCLUSIONS HJB can inhibit the proliferation of MDA-MB-231 and MDA-MB-468 cells， and induce their apoptosis.

Objective:To analyze the prognosis and influencing factors of different radiotherapy modes in patients with brain metastases from non-small cell lung cancer (NSCLC), and to explore the best benefit population with radiotherapy boost under different prognostic scores.Methods:634 patients with brain metastasis from NSCLC admitted to the Fourth Hospital of Hebei Medical University from 2013 to 2015 were analyzed retrospectively. According to different radiotherapy modes, they were divided into three groups: no radiotherapy group ( n=330), whole-brain radiotherapy group (WBRT)( n=127) and whole-brain radiotherapy combined with boost group (WBRT+ boost)( n=177). The intracranial progression-free survival (iPFS) and overall survival (OS) were calculated by Kaplan-Meier method. The multivariate prognostic factors were analyzed by the Cox models. Results:The median iPFS and OS of all patients were 6.9 months and 9.0 months, respectively. In the no radiotherapy, WBRT and WBRT+ boost groups, the 1-year iPFS was 15.1%, 16.3% and 40.2%( P=0.002), and the 1-year OS was 33.7%, 38.2% and 48.1%( P<0.001), respectively. Multivariate survival analysis demonstrated that different radiotherapy modes were the independent factors affecting iPFS and OS. Subgroup analysis revealed that for patients with 1-3 brain metastases, the 1-year OS and iPFS in the WBRT+ boost group were better than those of WBRT alone ( P=0.026, P=0.044) when GPA score was 2.5-4.0; the 1-year OS and iPFSin the WBRT+ boost group were better than those of WBRT alone ( P=0.036, P=0.049) when there was no targeted therapy; for patients with ≥4 brain metastases, the 1-year iPFS in the WBRT+ boost group was better than that of WBRT alone ( P=0.019, P=0.012) when GPA score was 2.5-4.0 and there was no targeted therapy. When the GPA score was 0-2 or there was targeted therapy, the 1-year OS and iPFS in the WBRT+ boost group were better than those of WBRT alone, but the difference was not statistically significant (all P>0.05). Conclusions:Radiotherapy can significantly improve the iPFS and OS of NSCLC patients with brain metastases. When the number of brain metastases is 1-3, GPA score is 2.5-4.0 or no targeted therapy, boost may improve the iPFS and OS; when the number of brain metastases is more than 4, GPA score is 2.5-4.0 or no targeted therapy, boost may only bring iPFS benefit; when GPA score is 0-2 or targeted therapy, boost may not benefit significantly.

Objective:To investigate the possible mechanism of mesenchymal stem cells (MSC) secreting hepatocyte growth factor (HGF).Methods:① C57BL/6 mouse mesenchymal stem cells (mMSC) were cultured in vitro, and mMSC with high expression of chemokine receptor 7 (CXCR7) were transduced by lentivirus plasmid. Blank control group and empty carrier control group were set at the same time. After 20 generations of cell culture, the transfection efficiency was identified by fluorescence microscopy and flow cytometry. The mRNA expression levels of CXCR7 in mMSC were detected by real-time fluorescent quantitative reverse transcription-polymerase chain reaction (RT-PCR). ② mMSC with passage number 4-6 were divided into MSC control group [MSC-blank group, 100 μg/L lipopolysaccharide (LPS) was added to wild-type MSC], highly expressed CXCR7 group (MSC-OE-CXCR7 group, 100 μg/L LPS was added to mMSC transduced by lentivirus plasmid with high expression of CXCR7), highly expressed CXCR7 control group (MSC-OENC-CXCR7 group, 100 μg/L LPS was added to mMSC transduced by no load lentivirus plasmid), CXCR4 inhibitor group (MSC-IE-CXCR4 group, 100 μg/L LPS was added to mMSC after 0.1 mg/L CXCR4 inhibitor TC14012 pretreatment for 24 hours), and CXCR4 inhibitor control group (MSC-IENC-CXCR4 group, 100 μg/L LPS was added to mMSC after DMEM culture medium with equal amount of TC14012 pretreatment for 24 hours). Cells in each group were collected after treatment with LPS, and mRNA expression of inhibitor of differentiation-1 (ID-1) was detected by RT-PCR. The cell supernatant was collected, and the levels of HGF were detected by enzyme linked immunosorbent assay (ELISA). Results:① The high expression of CXCR7 for mMSC which were transduced through lentivirus plasmid were successfully constructed detected by fluorescence microscope and flow cytometry. Compared with the blank control group, the expression of CXCR7 mRNA in the lentivirus with high expression of CXCR7 group was significantly increased (2 -ΔΔCt: 5.81±0.97 vs. 1.02±0.12, P < 0.05). There was no significant difference in CXCR7 mRNA expression between the empty carrier control group and the blank control group (2 -ΔΔCt: 0.95±0.22 vs. 1.02±0.12, P > 0.05). ②Compared with the MSC-blank group, high expression of CXCR7 in MSC-OE-CXCR7 group or inhibition of CXCR4 in MSC-IE-CXCR4 group could induce high expression of ID-1 mRNA in mMSC (2 -ΔΔCt: 5.56±0.66, 2.47±0.58 vs. 1.00±0.10, both P < 0.05) and increase HGF exocrine level (ng/L: 632.02±149.98, 217.21±40.53 vs. 108.53±24.62, both P < 0.05). However, there were no significant differences in ID-1 mRNA expression and HGF exocrine level of mMSC among MSC-OENC-CXCR7 group, MSC-IENC-CXCR4 group and MSC-blank group [ID-1 mRNA (2 -ΔΔCt): 1.01±0.27, 1.21±0.32 vs. 1.00±0.10, HGF (ng/L): 133.56±25.19, 107.11±25.30 vs. 108.53±24.62, both P > 0.05]. Conclusion:High expression of CXCR7 or inhibition of CXCR4 in MSC can increase the expression of ID-1 and promote the secretion of HGF, thus promoting pulmonary microvascular endothelial repair.

BACKGROUND:Symptomatic treatment is the main management strategy for patients with knee osteoarthritis at early metaphase. Some previous studies have demonstrated that neuromuscular electrical stimulation can al eviate pain in knee osteoarthritis patients. To date, this effectiveness, however, stil remains controversial.
OBJECTIVE:To assess the efficacy of neuromuscular electrical stimulation on pain in patients with knee osteoarthritis.
METHODRandomized control ed trials concerning the efficacy of neuromuscular electrical stimulation in the treatment of knee osteoarthritis were identified from the Medline database conducted up to July 3, 2014. These studies were selected independently by two reviewers according to the inclusion and exclusion criteria. Meta-analysis was performed using Revman5.2 software.
RESULTS AND CONCLUSION:A total of 5 randomized control ed trials consisting of 239 participants were included. The results of the meta-analysis indicate neuromuscular electrical stimulation has no significant impact on measure of pain in knee osteoarthritis patients in comparison to the blank control group [mean difference=-0.40, 95%confidence interval (-1.34-0.54), P=0.40]. Owing to the sample limitations of our study, it is hard for us to draw a conclusion that the application of neuromuscular electrical stimulation in managing pain in patients with knee osteoarthritis is of little significance. Further work based on large-sample and high-quality randomized control ed trials is needed to determine the role of neuromuscular electrical stimulation in pain in this population.

BACKGROUND:Reconstruction of the anterior cruciate ligament has become the gold-standard treatment for an anterior cruciate ligament rupture. Despite the popularity of the procedure, there remains a considerable amount of controversies over whether an autograft or anal ograft should be used for primary anterior cruciate ligament reconstruction. OBJECTIVE:To compare the clinical outcomes of al ograft and autograft in primary anterior cruciate ligament reconstruction. METHODS:Randomized control ed trials concerning autograft versus non-irradiated al ograft for anterior cruciate ligament reconstruction were identified from the PubMed/Medline database conducted up to July 12, 2014. These studies were selected independently by two reviewers according to the inclusion and exclusion criteria. Meta-analysis was performed using Revman5.2 software. RESULTS AND CONCLUSION:A total of 6 randomized control ed trials consisting of 858 cases were included, 441 cases in autograft group and 417 cases in non-irradiated al ograft. The results of the meta-analysis indicate no significant difference between autografts and non-irradiated al ografts in overal IKDC [relative risk (RR)=1.02, 95%confidence interval (CI) (0.99 to 1.06), P=0.21], degree of laxity [mean difference (MD)=-0.13, 95%CI (-0.29 to-0.02), P=0.09], Lachman test [RR=1.04, 95%CI (0.95 to 1.13), P=0.37], pivot shift test [RR=1.00, 95%CI (0.95 to 1.05), P=0.96], one-leg hop test [RR=1.01, 95%CI (0.96 to 1.06), P=0.77], Lysholm score [MD=-0.64, 95%CI (-1.45 to 0.17), P=0.12], Tegner score [MD=0.16, 95%CI (-0.16 to 0.47), P=0.34] and rate of postoperative complications [RR=1.42, 95%CI (0.67 to 3.04), P=0.36]. Therefore, in the meta-analysis, there is no significant difference between autograft and non-irradiated al ograft in clinical outcomes. However due to the limitations of our study, further work is needed to determine this conclusion.

Objective: To observe curative effect of radiofrequency catheter ablation (RFCA) and changes of cardiac function and left atrial diameter after operation in patients with atrial fibrillation (AF). Methods: A total of 28 AF patients with obvious clinical symptoms and without effective for drug therapy received RFCA in our hospital, their data were retrospectively analyzed. They received RFCA under guidance of three dimensional electro-anatomic mapping (EAM) system. Changes of cardiac function and left atrial diameter were evaluated by echocardiography and curative effect was evaluated by ambulary blood pressure monitoring before, three and six months after operation. Results: Pulmonary vein isolation rate was 100% in all patients. No severe complication occurred during or after operation. After six-month follow-up, 27 cases（96.4%）did not recur AF among the 28 patients; Compared with before operation, there were significant decrease in left atrial diameter [(37.3±4.8) mm vs. (33.6±4.5) mm] and significant increase in left ventricular ejection fraction [(59.8±8.7) % vs. (64.2±6.8) %] by echocardiography, P<0.05 both. Conclusion: Radiofrequency catheter ablation is safe and effective in treatment of atrial fibrillation, and there are significant improvements in cardiac function and left atrial diameter.

Objective To observe the clinical safety of dual axis rotational coronary angiography (DARCA) in the diagnosis of coronary artery disease in Chinese population.Methods From March to December in 2010,74 patients undergoing diagnostic DARCA were enrolled.The improved isocentering technique was adopted in 34 of the patients at the end of the study during DARCA.Blood pressure,heart rate and symptoms were recorded immediately before-and-after contrast injections.Contrast dose,radiation exposure and procedure time for DARCA were recorded.Continuous variable data were analyzed using Student＇s t test,if normality assumption was violated,rank sum test would be used.Categorical variables were analyzed using x2 test.Results ( 1 ) Clinical safety:There was no chest pain documented during or immediately post-injection for all patients.Only 1 patient ( 1 ％ ) had an attack of ventricular tachycardia immediately after the contrast injection and then relieved automatically.Pre and post-injection systolic blood pressure values of left coronary artery were statistically different [ ( 116 ± 20 ) mm Hg vs.( 111 + 18) mm Hg( 1 mm Hg =0.133 kPa),t =3.303,P =0.001 ],and heart rates differed,too [ 73 ( 65- 84)bpm vs.71(64-78) bpm,Z =-4.789,P =0.001 ],but that imposed no clinical significance.(2)Contrast dose,radiation dose and procedure time:The mean contrast utilization,radiation dose and procedure time for DARCA were 28 (25-34) ml,8979 ( 6733-12 363 ) mGycm2 and 200 (164-270) s.Compared with conventional DARCA,improved isocentering technique during DARCA had less radiation exposure and procedure time in left coronary artery angiography and the whole coronary artery angiography [ left coronary artery angiographic radiation exposure:4004 (2932-5772) mGycm2 vs.5808 (4798- 8838) mGycm2,Z =-3.471,P =0.001 ;total radiation exposure:(8116 +2493) mGycm2 vs.( 11 371 ±4122) mGycm2,t =-4.176,P =0.001 ; left coronary artery angiographic procedure time:120 ( 80-180)s vs.150(126-214) s,Z =- 2.836,P =0.005; total procedure time:180 (139-240) s vs.220( 186-308 )s,Z =-3.004,P =0.003 ],but there was no statistically difference in contrast utilization [30(25-35) ml vs.27(25-34)ml,Z=-0.906,P=0.365].Conclusion This study demonstrates clinical safety of DARCA in the diagnosis of coronary artery disease in Chinese population.Compared with conventional isocentering technique of DARCA,improved isocentering technique can significantly reduce radiation exposure and procedure time on the basis of simplified operation,and replace the conventional isocentering technique,but randomized double-blind controlled studies should be conducted.

Objective To compare the the effect of PCI on short term (3 months) left ventricular diastolic function in the old patients with acute myocardial infarction (AMI) (≥ 65 years old). Methods 40 old AMI patients were divided into PCI group and non-operation group (pure medicine treatment group). The changes of left ventricular diastolic function were observed 3 months after operation which were compared with that before treatment so to define the effect of PCI. The major indexes were LA, LV, LVEF, E/A, and S/D. Results LA and LV in PCI group were not significandy changes but they were increased in non-operation group (P < 0.05). Although the left ventricular diastolic function in both group was continually impaird,it was better in PCI group than in non-operation greup(P <0.05). Conclusion PCI can prolong the progress of left heart remodeling and the excerbation of left ventricular diastolic function in old AMI patients.