Objective To establish RNase L gene knockout HEK 293 cell lines using CRISPR/Cas9 system.Methods Small guide RNA ( sgRNA) sequences of human RNase L were designed and sgRNAs were inserted into pCas-Guide and pCas-guide RNA(gRNA) vectors were obtained.The donor DNA sequences of the homologous arm were designed for RNase L knockout .In the presence of the right homologous arm , the resistance gene of hygromycin B and the left homologous arm as templates of homology-directed repair , the donor DNA template was amplified by overlopping PCR and cloned into the pBackZero-T expression vector and pBackZero-T-RNase LK vector was obtained .The pCas-gRNA vector and pBackZero-T-RNase LK vector were co-transfected into HEK293 cells to establish the stable expression cell line of RNase L gene knockout .Cells were cultured with hygromycin B , while Western blotting and DNA sequencing were used to analyze the gene of RNase L knockout from genome .Results and Conclusion The pCas-gRNA vector and pBackZero-T-RNase LK vector were successfully constructed.Five RNase L gene knockout HEK293 cell lines were generated,contributing to the study of the biological function and molecular mechanism of RNase L .

Objective To develop and optimize a new method to extract miRNAs from plasma.Methods miRNAs were extracted from plasma by mixing it with the extraction solution that contained surfactant and by heating .Then the ribonuclease inhibitor was added into the extraction to prevent RNAs from degradation .The expression level of each miRNA was detected by real-time quantitative PCR in oder to evaluate the feasibility of this method .Results A method which extracted miRNAs from plamsa in just one step was established .The specificity , reproducibility and stability of this method have been demonstrated by real-time quantitative PCR .Conclusion The one-step method is simple , inexpensive , and plasma-saving.It seems like a new method for clinical examination of miRNAs from plasma .

Objective This study was to explore the expression and significance of HMA2 in bladder cancer , analyze its correlations to clinicopathologic and recurrence of bladder cancer. Methods The expression of HMGA2 protein in 148 specimens of bladder cancer and 30 specimens of normal bladder tissues was detected by immunohistochemtry, its correlations to clinicopathologic features was analyzed. Results There was no expression of HMGA2 protein in normal bladder tissues,while the expression level of HMGA2 protein was getting higher with the increase of tumor pathology grade and stage. The positive rate of HMGA2 protein was 21.3% in G1 bladder cancer, 60. 3% in G2 bladder cancer, 82.1% in G3 bladder cancer, its difference is significant (P＜0. 001). It was significantly lower in non-muscle invasive bladder cancer than in muscle invasive bladder cancer (43.3% vs 72. 7%, P=0. 003). The patients were followed up for 2～95 months, patients of recurrence was 64,HMGA2 protein expression was significantly higher in patients with recurrence than with non-recurrence (54.7% vs 25.0%, P=0. 007). Conclusions The expression of HMGA2 protein was highly in bladder cancer, the positive rate of HMGA2 protein expression was related with classification,TMN stage and recurrence, but not with sex, age, tumor number (P＞0. 05). The detection of the expression of HMGA2 protein is in favor of diagnosis and prognostic evaluation of bladder cancer.

Abstract In April 2021, type Ⅰ vaccine-derived poliovirus (VDPV) was detected from two fecal samples of a male infant with acute flaccid paralysis (AFP) in Zhejiang Province when he was admitted to the Children's Hospital Affiliated to Fudan University in Shanghai, with 12 and 14 nucleotide mutations in the VP1 region, respectively. The case had a history of immunization with three doses of poliovirus vaccines, and grade Ⅲ proximal muscle strength and grade Ⅱ distal muscle strength of the right lower limb. After symptomatic treatment, the activity of the right lower limb and the muscle strength was significantly restored, thus he was discharged. VDPV was not detected from subsequent (the 8th to 12th) fecal samples of the case and fecal samples of close contacts. No similar cases were found in medical institutions in the county, surrounding areas, neighboring villages or towns. Since the case did not exhibit clinical symptoms of poliomyelitis caused by VDPV, poliomyelitis was excluded, and the case was diagnosed with hemophilia type A based on the epidemiological investigation, laboratory tests, and the history of poliomyelitis vaccination. This event involved cross-provincial (municipal) cooperation and was responsed promptly, preventing further spread of the virus. It suggested that the sensitivity of the AFP case surveillance system should be maintained, environmental monitoring methods should be increased, and the poliomyelitis vaccination should be promoted to prevent the spread of the virus.

Aim TostudytheeffectsofCuO,ZnOand TiO2 nanoparticles on the viability and metastatic po-tential of EC-9706 and EC-109 esophageal squamous carcinomacelllineinvitro.Methods Characteristics of CuO,ZnO and TiO2 nanoparticles were detected u-sing transmission electron microscope (TEM)and dy-namic light scattering (DLS ).EC-9706 and EC-109 cells were treated with different concentrations of CuO, ZnO and TiO2 (5 ~80 mg · L-1 ).The cell prolifera-tion was analyzed by MTT assay.The cell cycle and apoptotic rates were determined by flow cytometry (FCM).The cell invasion was assayed in Transwell chambers.The expression of Bcl-2 and caspase-3 pro-tein in cells was detected by Western blot method.Re-sults CuO,ZnOandTiO2nanoparticleswerespheri-cal with primary particle size 12,20. 6,12 nm.The particles were agglomerated in water and cell culture medium with negative charge.CuO and ZnO nanoparti-cles induced decreases in EC-9706 and EC-109 cell vi-ability dose-dependently.After exposed to increasing concentrations of CuO and ZnO nanoparticles,the cell cycle analysis revealed a decreasing proportion of cells in G2/Mand S phase,and up-regulation of the cells in G0/G1 phase.Apoptotic cells also increased along with decreased cell invasion upon CuO and ZnO treatment. Nanoparticles treatment after 48 h, the activated caspase-3 expression quantity increased significantly and the Bcl-2 expression quantity decreased obviously (P<0. 05 )compared with control group.TiO2 nanop-articles had no obvious effect on the EC-9706 and EC-109 cell proliferation,cell cycle,apoptosis and inva-sion.Conclusion ComparedwithTiO2,CuOand ZnO nanoparticles can inhibit EC-9706 and EC-109 cell viability and metastatic potential,the mechanism of action involves cell cycle arrest in G0/G1 phase and apoptosis.These findings can help the development of nanoparticles as anti-cancer therapeutics for esophageal cancer.

Objective· To evaluate the safety of neoadjuvant therapy which was constituted by docetaxel based systemic chemotherapy and maximal androgen blockage for patients with locally advanced prostate cancer and to summarize the related adverse events and clinical managements.Methods· From June 2015 to February 2017,the clinical data of 55 patients undergoing neoadjuvant chemotherapy combined with complete androgen deprivation were retrospectively reviewed.The patients were given docetaxel and prednisone as DP regimen every 3 weeks and LHRH analogues with bicalutamide as maximal androgen deprivation for a total of 4 cycles.All treatment-related adverse events were observed and then recorded.Results· Two cases with liver function impairment after 2 cycles of treatment were withdrawn from the study.No severe allergic reactions occurred during neoadjuvant therapy.The most common adverse events were hematologic toxicity,while 23.6％ of patients had grade Ⅲ-Ⅳ neutropenia,and about 12.7％ had anemia.Due to a relatively short course of treatment,the skin or mucous damage,peripheral neurotoxicity and fluid retention were rare.However,hot flash,male breast development as well as erectile dysfunction were very frequently observed due to maximal androgen deprivation.The majority of these adverse events were relieved by symptomatic and supportive treatment.Conclusion · After strict selection,4 cycles of neoadjuvant chemotherapy combined with total androgen blockade could be well tolerated by the patients with high-risk locally advanced prostate cancer.Even though the adverse events were controllable,they still need to be closely monitored during treatment in order to reduce the incidence.In addition,the very low testosterone level associated endocrinal metabolic disorders caused by complete androgen deprivation were also of great concern.

Nicotinic acetylcholine receptors (nAChRs) ,also known as neuronal nicotinic receptors ,are widely expressed throughout the central and peripheral nervous system .nAChRs play crucial roles in pain signaling .Recently ,agonists that tar-get specific nAChR subtypes have shown substantial efficacy in a wide range of acute and chronic pain models ,contributing to developing novel analgesic drugs with low drug dependence .Positive allosteric modulators offer new approaches for increasing the potency and therapeutic window of these drugs .This review summarizes the advances in nicotinic acetylcholine receptors as analgesic targets and anticipate the future directions in this field .

Objective: To investigate the effects of emodin on lipopolysaccharide (LPS)-induced inflammation and damage in HK-2 cells. Methods: The HK-2 cells were divided into blank group, emodin group, LPS group, and LPS+emodin group. The emodin group was treated with culture medium containing 0.5 μg/ml emodin, the LPS group was treated with culture medium containing 10 μg/ml LPS, the LPS+emodin group was treated with culture medium containing both 0.5 μg/ml emodin and 10 μg/ml LPS, and the blank group was cultured with fresh medium. Twelve hours later, HK-2 cells from each group were harvested for the analysis. The mRNA levels of oligonucleotide receptor protein 3 (NLRP3), IL-1β, IL-18, TNF-α, and neutrophil gelatinase-associated lipocalin (NGAL) in each group were determined by RT-PCR. Results: Compared to the blank group, the mRNA levels of NLRP3 (2.74 ± 0.38 vs. 1.00 ± 0.14), IL-1β (2.40 ± 0.33 vs. 1.00 ± 0.19), and IL-18 (3.00 ± 0.42 vs. 1.00 ± 0.07), NGAL (31.73 ± 3.41 vs. 1.00 ± 0.07), and TNF-α (9.73±1.60 vs. 1.00 ± 0.11) significantly increased by LPS stimulation (P<0.05). Compared to the LPS group, the mRNA expression of NLRP3 (1.98 ± 0.24 vs. 2.74 ± 0.39), IL-1β (1.54 ± 0.24 vs. 2.40 ± 0.33), and IL-18 (2.09 ± 0.53 vs. 3.00 ± 0.42), NGAL (16.76 ± 1.72 vs. 31.73 ± 3.41) and TNF-α (9.73 ± 1.60 vs. 4.65 ± 1.09) in HK-2 cells of LPS+emodin group significantly decreased (P<0.05). Conclusions The emodin protects HK-2 cells from LPS-induced damage and inflammation by inhibiting NLRP3 inflammasome activation.

OBJECTIVETo study the effect and safety of the Chinese-made inflatable penile prosthesis in the treatment of erectile dysfunction (ED).

METHODSAccording to the anatomy and physiology of Chinese men, an inflatable penile prosthesis was developed in China, consisting of an innovated pump, a reservoir, and a pair of penile cylinders, which were connected by tubes to form a complete system. It was authorized to be used clinically by State Food and Drug Administration (SFDA). Forty-five cases of organic ED were selected for penile prosthesis implantation. Penoscrotal approach was used to implant the cylinders into the corpus cavernosum, the pump into the scrotum and the reservoir into the prevesical space.

RESULTSSurgical problems and mechanical failures were not found in the subjects. Post-operative complications occurred in 3 cases (6.6%), but did not affect the intercourse. Satisfactory intercourses were achieved 10-12 months later and the intercourse duration averaged about (20 +/- 6) min.

CONCLUSIONThe Chinese-developed inflatable penile prosthesis was safe and effective for patients with ED, with low rate of mechanical failures. Its long term effect has yet to be further studied.

Adult ; Erectile Dysfunction ; surgery ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Penile Implantation ; Penile Prosthesis ; Prosthesis Design

Objective:To investigate the associations of articular depression depth (ADD) and tibial plateau widening (TPW) by pre-operative CT measurement with incidence of lateral meniscal tear in patients with Schatzker type Ⅱ tibial plateau fracture.Methods:Included in this retrospective study were 131 patients who had been admitted to Emergency Center of Trauma, The Third Hospital Affiliated to Hebei Medical University from January 2016 to January 2020 for Schatzker type Ⅱtibial plateau fractures. They were 88 males and 51 females, aged from 18 to 60 years (average, 41.5 years), with 74 right and 57 left sides injured. All patients were treated with closed reduction and internal fixation assisted by bidirectional traction. Arthroscopy was used to detect the status of lateral meniscus immediately after closed reduction and internal fixation of the fracture fragments. Furthermore, patients were divided into 2 groups according to the integrity of lateral meniscus: meniscal tear group ( n=70) and tear-free group ( n=61). The 2 groups were compared in terms of age, gender, body mass index(BMI), injury side, time interval from injury to surgery, TPW and ADD. The receiver operating curve (ROC) was drafted to calculate the cut-off values of TPW and ADD in complication of lateral meniscal tear in patients with Schatzker type Ⅱ tibial plateau fracture. Results:The overall incidence of lateral meniscal tear in this cohort was 53.4% (70/131). There was no statistically significant difference in terms of age, gender, injury side, BMI or time interval from injury to surgery between the 2 groups ( P>0.05); TPW and ADD were significantly higher in the meniscal tear group than in the tear-free group ( P<0.05). To predict lateral meniscal tear in patients with Schatzker type Ⅱtibial plateau fracture, the area under ROC was 0.656 (95% CI: 0.562 to 0.750, P=0.002) for TPW and 0.709 (95% CI: 0.619 to 0.800, P<0.001) for ADD, respectively; the cut-off values of TPW and ADD were 4.3 mm and 6.1 mm. Conclusion:TPW and ADD may be effective predictors for prediction of lateral meniscal tear in patients with Schatzker type Ⅱ tibial plateau fracture.