Objective To identify genetic regulators of ionizing radiation(IR)sensitivity through clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated nuclease 9(Cas9)genome-wide screening.Methods A specialized single guide RNA(sgRNA)library was developed targeting 987 stress-response regulatory genes identified through Kyoto Encyclopedia of Genes and Genomes(KEGG),Reactome and gene ontology(GO)databases,followed by construction of sgRNA plasmids and packaging into a lentiviral library.Using colon adenocarcinoma Caco-2 cells as a model system,the Cas9-stable transgenic line was established via lentiviral transduction and antibiotic selection.Library-transduced cells underwent puromycin selection,followed by γ-irradiation(dose optimized via preliminary experiments).Post-irradiation phenotypic selection was conducted after 14 days,with subsequent next-generation sequencing of surviving cell populations to identify enriched/depleted sgRNAs.Candidate genes were functionally validated through orthogonal assays:cell counting kit-8(CCK-8)proliferation analysis,clonogenic survival assays and flow cytometric quantification of reactive oxygen species(ROS)and apoptotic markers.Results The optimized screening platform identified 281 radiation response genes(165 radiosensitive and 116 radioprotective candidates).Functional validation revealed Bcl2-associated athanogene 3(BAG3)knockdown significantly enhanced radioresistance.Proliferation was increased 1.2-1.5 fold,clonogenic survival improved 1.5-2.0 fold,and ROS was reduced by 13％-25％coupled with 32％-73％apoptosis attenuation compared to control groups.Conclusion The integrated CRISPR/Cas9 screening platform effectively identifies novel radiation response regulators,as evidenced by the discovery of BAG3 as a critical radiosensitivity determinant.This high-throughput functional genomics approach provides a robust methodology for systematically mapping molecular determinants of cellular radiation response,with potential applications in both mechanistic studies and therapeutic target discovery.

Persistent left superior vena cava(PLSVC)is a common congenital anomaly of systemic venous drainage,often draining into the right atrium without the need for special treatment.Sometimes,PLSVC drains into the left atrium,creating a right-to-left shunt,leading to reduced blood oxygen saturation and paradoxical embolism,requiring intervention.Traditional surgical ligation of PLSVC is the conventional approach for managing abnormal shunting,but it is associated with significant trauma and carries the risk of damaging the phrenic nerve.Here,we present a case of a patient with right heart dysfunction due to an untreated PLSVC-left atrium communication after corrective surgery for complex congenital heart disease,resulting in left-to-right shunting postoperatively.The patient was successfully treated by using a Plug vascular occluder via a transseptal approach to occlude the PLSVC.To our knowledge,this is the first report of successful closure of the left-to-right shunting through the heart chambers via a transseptal approach,indicating that interventional occlusion is an ideal management approach.

Low-level patent foramen ovale nonocclusion(PFO)is a rare type of PFO in which the PFO opening is low during transcatheter closure of PFO and the distance between the PFO left atrial opening and the root of the septal side of the mitral valve is less than 9 mm,and the smallest model of the current double-disk PFO occluder(18/18)commonly used in clinical practice for low-level PFOs can touch the mitral valve,resulting in increased risk of mitral regurgitation or leaflet abrasion.The risk of mitral regurgitation or leaflet abrasion is increased,and transcatheter closure of PFO procedure can only be abandoned when encountered intraoperatively.In this article,we present a case of successful transcatheter closure of a low-level PFO using the Amplatzer ADOⅡ occluder,which provides new ideas and strategies to deel wtih this rare type of PFO.

This paper explored the background, framework and approach, contents and implementation of WHO Rehabilitation in Health System using approaches of ICF and WHO Handbook for Guideline Development. The actions and significances of implementations of seven recommendations and one good practice statements on assistive products had been discussed.

OBJECTIVE: To construct a decellularized matrix of human fatty liver as the scaffold for three-dimensional (3D) culture of hepatocarcinoma cells. METHODS: Human fatty liver decellularized matrix (hFLM) was prepared by repeated freezingthawing, perfusion with gradient SDS and 1% Triton X-100 through the portal vein and hepatic artery, and repeated agitation with Triton X-100. HepG2 cells were cultured in the prepared hFLM, and the cell survival, morphology, proliferation and cellular expressions of the adhesion molecules were detected. RESULTS: The decellularization procedure shortened the time for scaffold preparation and preserved the 3D ultrastructure and the composition of the extracellular matrix. HepG2 cells cultured in hFLM scaffold maintained proliferation for up to 15 days and showed a growth pattern with a long lag phase and a slow growth rate, which was similar to the growth pattern . The cultured HepG2 exhibited a low expression of E-cadherin and a high expression of vimentin, which was consistent with the xenograft but opposite to 2D cultured cells. However, the lack of adequate nutrient transport in this hepatocarcinoma cell model led to a slowdown of cell proliferation in the later stage. The PCNA index of HepG2 cells cultured in hFLM was lowered by 29.3% on day 12 as compared with that on day 6. CONCLUSIONS We established a new protocol for preparing hFLM and confirmed the feasibility of constructing hepatocarcinoma cell models using the hFLM scaffold.
Carcinoma, Hepatocellular ; Extracellular Matrix ; Fatty Liver ; Humans ; Liver Neoplasms ; Tissue Engineering ; Tissue Scaffolds

BACKGROUND: Proximal femoral nail anti-rotation is widely used to treat various intertrochanteric fractures. Although its operation trauma is small, and the blood loss of perioperative period is still large. Tranexamic acid has been gradually used to reduce the bleeding of intertrochanteric fracture. The effectiveness and safety of reducing blood loss during perioperative period were not reported. OBJECTIVE: To explore the safety and efficacy of tranexamic acid on perioperative blood loss in patients with intertrochanteric fracture undergoing proximal femoral nail anti-rotation. METHODS: One hundred and eight patients with intertrochanteric fracture undergoing proximal femoral nail anti-rotation were selected from First Affiliated Hospital, Guangzhou University of Chinese Medicine between January 2015 and January 2017. Among all the subjects, 52 patients who received the operation before January 2016 served as the control group and 56 patients who received the operation after January 2016 were selected as the treatment group. Half an hour before operation, patients in the treatment group received 1 g tranexamic acid dissolved in 250 mL normal saline by intravenous dropping; patients in the control group just received 250 mL normal saline by intravenous dropping. The bleeding volume, blood transfusion volume, hemoglobin, hematocrit, coagulation index, D-dimer levels and complications were compared between the two groups. RESULTS AND CONCLUSION: (1) During perioperative period, actual blood loss, intraoperative blood loss, dominant blood loss, recessive blood loss, volume of drainage, blood transfusion volume and blood transfusion rate were lower in the treatment group than in the control group (P < 0.05). (2) There was no statistically significant difference in the hemoglobin and hematocrit between the two groups before operation (P > 0.05). The hemoglobin and hematocrit of the two groups gradually decreased after the operation, and there was a slight improvement in the fifth day after surgery. At postoperative 2 hours, 1, 3 and 5 days, the hemoglobin and hematocrit of the treatment group were higher than in the control group (P < 0.05). At preoperation and each time point postoperation, prothrombin time, activated partial thromboplastin time, and fibrinogen levels were not statistically significant between the two groups (P > 0.05). Postoperative D-dimer levels in the two groups were significantly higher than preoperation, and there was a return on the fifth day. There was no statistically significant difference between groups at preoperation and each time point of postoperation (P > 0.05). (3) The results suggest that the tranexamic acid can effectively reduce the dominant and recessive blood loss in patients with the intertrochanteric fracture, and it is safe and effective.

Objective :To explore therapeutic effect and safety of irbesartan hydrochlorothiazide (I+H) tablet combined am-lodipine besylate on young and middle-aged patients with hypertension .Methods : A total of 138 young and middle-aged pa-tients with hypertension ,who were treated in our hospital from Jan 2015 to Feb 2017 ,were selected .Patients were ran-domly and equally divided into group A (received I+ H therapy) ,group B (received amlodipine treatment ) and group C (received I+ H+ amlodipine treatment ) ,all groups were continuously treated for three months .Systolic blood pressure (SBP) ,diastolic blood pressure (DBP) and heart rate (HR) before and after treatment ,total effective rate and incidence rate of adverse reactions were measured and compared among three groups .Results :Compared with before treatment ,after treatment ,there were significant reductions in SBP ,DBP and HR in three groups , P＜0.05 or ＜0.01. Compared with group A after treatment ,there was significant reduction in SBP [ (133.1 ± 6.2) mmHgvs.(128.9 ± 6.0) mmHg vs. (122.0 ± 3.8) mmHg] in group B and C ,and that of group C was significantly lower than that of group B ,P＜0.01 all ;compared with group B after treatment ,there was significant reduction in DBP [ (91.7 ± 6.7) mmHg vs.(87.5 ± 4.8) mmHg vs.(79.3 ± 3.0) mmHg] in group A and C ,and that of group C was significantly lower than that of group A ,P＜0.01 all.Compared with group A and B ,there was significant rise in total effective rate (78.3% vs.73.9% vs.97.8%) in group C , P＜0.01 all.There was no significant difference in incidence rate of adverse reactions among three groups , P＝0.876 .Conclusion : Irbesartan hydrochlorothiazide tablet combined amlodipine besylate tablet can significantly improve blood pressure in young and middle-aged patients with hypertension , Its effect is better than that of two drugs alone use . Which is safe and reliable ,and is worth extending .

Objective:To investigate the effect of inhibiting of HOXB7 gene expression on proliferation and apoptosis of colon cancer cells.Methods:The synthetic negative control siRNA (negative control group) and HOXB7-siRNA (HOXB7 transfection group) were transfected into human colon cancer SW480 cells by LipofectamineTM2000 liposome mediated method,untreated cells as blank group.RT-PCR and Western blot were used to detect the mRNA and protein expression of HOXB7 after transfected 48 h respectively;cell proliferation was detected by CCK8 assay after transfected 24,48,72,96 h;cells apoptosis was detected by flow cytometry after transfected 48 h;the expression of apoptosis related proteins Bcl-2,Bax and Notch1 signaling pathway Notch1 and Hes1 were detected by Western blot.Results:The mRNA and protein expression of HOXB7 in HOXB7 transfected group was significantly lower than that in blank group(P<0.05);OD value was no statistical significance in the three groups after transfected 24 h(P>0.05), while after transfected 48,72,96 h,compared with the control group,OD value in HOXB7 group was significantly decreased(P<0.05);compared with the blank group,the apoptosis rate in HOXB7 transfection group increased significantly,and the expression of Bcl-2, Notch1 and Hes 1 proteins was down regulated,and the expression of Bax protein was up-regulated(P<0.05).Conclusion:RNA inter-ference in the expression of HOXB7 gene in colon cancer can inhibit the proliferation of cancer cells and induce apoptosis by inhibiting of the Notch1 signaling pathway.

OBJECTIVETo develop a method for preparing a decellularized scaffold based on human liver tissue.

METHODSA surgical specimen of the left lateral lobe of the liver was obtained from a patients with hepatic hemangioma. The decellularization process was performed by repeated freezing-thawing, sequential perfusion with 0.01% SDS, 0.1% SDS and 1% Triton X-100 through the portal vein, and sterilization with peracetic acid. L-02 cells were then engrafted onto the decellularized liver scaffold.

RESULTSHE staining, DAPI staining and scanning electron microscopy all verified the absence of residual cellular components in the decellularized scaffold. The residual DNA content in the decellularized scaffolds was 25.3∓14.6 ng/mg (dry weight), which was less than 1% of the total DNA content in a fresh human liver. Immunohistochemistry demonstrated that type I and IV collagens, fibronectin and elastin were all retained in the scaffold. The engrafted L-02 cells survived well on the scaffold with active proliferation and expressed albumin and G6pc.

CONCLUSIONIt is feasible to prepare decellularized scaffolds using surgical specimens of human liver, which can be a new approach to constructing a tissue-engineered liver for clinical purposes.

Objective To explore the electrophysiological charaterstics of upper extremities nerves on the patients with Hirayama disease (HD),amyotrophic lateral sclerosis (ALS),and distal cervical spondylotic amyotrophy (DCSA).Methods The data of electrophysiological examination of the upper limbs of 87 patients with HD,83 with ALS and 28 with DCSA were reviewed retrospectively.Seventy-two patients with HD among 87 had unilateral upper limb's amyotrophy and the other 15 ones had bilateral amyotrophy.There were 30 patients had unilater upper limb's amyotrophy and 53 ones had bilateral amyotrophy from the group of patients with ALS; 20 patients with DSCA were affected unilaterally and 8 ones were bilaterally affected.Results Compound muscle action potential (CMAP) evoked by ulnar stimulation had a lower ampititude compared with that evoked by median stimulation in HD patients.In ALS cases that was just the opposite.However,the CMAPs were similar in DCSA cases.The mean ratio of CMAP amplitude by ulnar stimulation to by median stimulation was 0.58±0.40 in HD group; 2.28±1.25 in ALS and 1.31±0.63 in DCSA.The differences in the three groups were statistical significance.The U/M CMAP ratio was less than 0.6in 62 patients with HD,3 with ALS and 1 with DCSA,and more than 1.7 in 73 cases (57 ALS,12 HD and 4 DCSA).Conduction velocities (CV) of the sensory and motor nerves,the amplitude of the sensory nerve action potential in bilateral limbs,and the CMAP amplitude of the unaffected limb were normal in all cases.Conclusion This study could concluded that the severity of amyotropy in hypothenar mucles were higher than that in thenal muscles in patients with HD; there was just opposite in ALS cases and similar in DSCA.