Objective To knock out p21 gene in human malignant rhab doid tumor(MRT)cell line G401 by using CRISPR/Cas9 genome engineering technology. Methods The expression of p21 was detected by reverse transcription quantitative PCR (RT-qPCR) and Western blot assay in several MRT cell lines. The guide RNA was designed by targeting the third exon of p21 gene,which encoded its home domains, and then subcloned into lentiCRISPR v2 vector and validated sequencing. The validated plasmids were further used to package and produce the lentivirus in 293T cells, and the G401 cells were infected, then puromycin was used to screen positive cells, and the clusters of G401 monoclonal cells, were obtained by selecting monoclonal cells and culturing under the microscope. The RNA and protein of new clonal cell line were extracted, and RT-qPCR and Western blot assay were applied to confirm whether p21 was successfully knocked out. Results The p21 was highly expressed in MRT tumor cells. The CRISPR/Cas9 lentivirus plasmids, targeted p21 gene were successfully constructed. Compared with negative control group,the expression of p21 was not detected in G401 monoclonal cells, which were successfully screened. Conclusion In view of the difficult transfection of cells such as G401, p21 knockout stable cell line has been successfully constructed by using CRISPR/Cas9 system, which lays the foundation for further study of the mechanism of p21 in MRT tumors .

Anal malignant melanoma is a rare disease with high malignancy and poor prognosis. This paper reviews the relevant literature of the diagnosis and treatment of a case of anal melanoma.

Lipoma of hemangiopericytomas (LHPC) is one ofrare soft-tissue tumors that grow slowly and occur in deep soft tissues. The classical histological morphology of LHPC is circularor spindle undifferentiated tumor cells that grow surrounding the thin-walled branching blood vessels. The clinical and pathological features of one patient with LHPC are retrospective analyzed to strengthen the understanding of LHPC.

Anti-S antibody is rare and irregular antibody in MNS blood group system. One patient was found positive when doing antibody screening experiment before coronary artery bypass grafting. This is the first case of serum IgG-anti-S in our laboratory. S-antibody screening test and irregular antibody identification are important before blood transfusion, which can reduce the transfusion reaction.

In recent years, microRNAs (miRNAs) have been found to be one of the key factors of post transcriptional gene regulation, which are involved in occurrence and development of many diseases. Peritoneal dialysis (PD) has become an effective alternative treatment approaches for patients with end stage renal disease (ESRD). Peritoneal fibrosis is one of the most important factors leading patients to withdraw from long-term PD, hence restricts the application and development of PD. MicroRNAs are closely related to the development of peritoneal fibrosis. This article reviews the role of miRNAs in the pathogenesis and treatment of peritoneal fibrosis.

Objective To investigate the effect of surgical treatment of adult mandibular condylar fractures (including intracapsular fracture). Methods Thirty-two patients (33 sides) with condylar fractures underwent open reduction and rigid fixation. Six patients with intracapsular condylar fractures were treated with two 18-mm positional screws through a preauricular approach. Six patients (7 sides) with condylar neck fractures were rigidly fixed by 1 mini-plate via a retromandibular transparotid approach. Twenty patients with subcondylar fractures were operated and fixed by two titanium plates using an angular approach. Results The mean follow-up period was 13.5 months, and the mean maximum mouth opening was 37.5 mm by the last visit. All patients acquired good occlusal relationship and mandibular symmetry. Seven patients (21.9%) experienced transient palsy of the branches of the facial nerve, and recovered completely after three months. One patient developed a salivary fistula, and healed after two weeks of gauze compression. No permanent deficit of any facial nerve branch was observed. No patient showed condylar head resorption. Conclusion Appropriate surgical approaches and fixation methods for different types of condylar fractures are the key factors to achieve reliable clinical results.

Objective To explore the clinical efficacy and outcomes of thrombus aspiration in ST-elevation myocardial infarction (STEMI) patients undergoing percutaneous coronary intervention (PPCI). Methods A total of 664 cases of STEMI patients undergoing PPCI from Tianjin Chest Hospital from January 2013 to March 2015 were retrospectively analyzed. Patients were divided into two groups:primary PCI after thrombus aspiration group and conventional primary PCI without thromobus aspiration group. The base-line clinical characteristics, and the procedure of percutaneous coronary intervention were similar between two groups. The TIMI flow, LVEF, one-month outcomes and 12-month outcomes were compared between two groups. Results There were significant higher TIMI flow, LVEF in primary PCI after thrombus aspiration group than those of conventional primary PCI without thromobus aspiration group (P<0.05). At one-month follow-up, there were no significant differences in major adverse cardiac events (MACE) between the two groups. At 12-month follow- up, there were no significant differences in cardiovascular death, recurrent myocardial infarction and new hospitalization between two groups. The incidence of recurrent angina was significantly higher in non-aspiration group than that in aspiration group (P<0.05). There were the better survival functions without MACE in aspiration group than that of control group. Conclusion Thrombus aspiration, as an adjunctive method to primary PCI for STEMI, may improve TIMI flow, have beneficial effects on LVEF and reduce the incidence of recurrent angina at 12-month follow up.

Objective To investigate the influence and clinical significance of single and dual-chamber pacing on central aortic pressure (CAP) and augmentation index (AI) in non-smoking individuals. Methods Totally, 83 non-smokers with pacemaker-implanted were consecutively enrolled in this study, and they were divided into three groups:dual-chamber pacemaker group (DDD, n=35), single-chamber pacemaker group (VVI, n=33) and control group (n=15). Heart rate (HR), CAP, AI, systolic blood pressure (SBP) and diastolic blood pressure (DBP) were measured in three groups of patients. Finally, DDD pacing mode was turned into VVI pacing mode in patients of DDD group and the indexes were measured again. All of the indexes were recorded and analyzed. Results There were no significant changes in baseline characteristics and laboratory data between three groups (P>0.05). Left atrial diameters were significantly higher in VVI group than those of control group (P<0.05). Values of CAP were significantly higher in DDD group than those of control group and VVI group (P<0.05). Values of AI, corrected AI (AIc) and brachial BP were significantly higher in DDD group than those of VVI group (P<0.05). Values of CAP and brachial BP were significantly lower in VVI group than those of control group (P<0.05), while no significant changes were found in AI and AIC between VVI group and control group (P>0.05). All of these indexes (CAP, AI and brachial BP) were significantly reduced after the pacing mode was changed (P<0.05). Conclusion In non-smokers, dual-chamber pacing mode can increase CAP and AI.

Objective To investigate the relationship between plasma homocysteine (Hcy), Kv1.3 channel and cardiac troponinI (cTnI) in patients with acute ST-segment elevation myocardial infarction (STEMI). Methods According to the level of Hcy, 80 STEMI patients were divided into STEMI with Hhcy group (Hcy > 15 μmol/L, n=41) and control group (STEMI group, Hcy≤15μmol/L, n=39). The Hcy, blood lipid and cTnI were detected with automatic biochemistry analyzer, respectively. Peripheral lymphocytes were isolated by ficoll density gradient centrifugation. Real-time PCR was used to detect mRNA expression of Kv1.3, and Western blot assay was used to detect protein expression of Kv1.3. Results cTnI concentrations were obviously higher in STEMI with Hhcy group than those in STEMI group (μg/L:22.997 ± 5.880 vs. 12.881 ± 6.343;P<0.01). Multiple linear regression analysis showed that age, gender, hypertension, diabetes mellitus, smoking, family history, total cholesterol (TC), triglyceride (TG), high density lipoprotein-cholesterol (HDL-C) and low density lipoprotein-cholesterol (LDL-C) had no obvious influence on Hcy (P>0.05). The relative expression levels of Kv1.3 mRNA and protein were significantly higher in STEMI with Hhcy group (1.35±0.14, 0.85±0.12) than those in STEMI group (1.00 ± 0.07, 0.64 ± 0.05, P<0.05). Moreover, there was a positive relation between Hcy level and the mRNA and proteinexpression of Kv1.3 channel (r=0.299, r=0.542, P<0.05). There was a positive relation between protein expression levels of Kv1.3 channel and cTnI (r=0.644, P<0.05). Conclusion Our results support that Hcy could exacerbate the concentration of cTnI through playing an important role in the Kv1.3 mRNA and protein expression in lymphocytes.

Objective To investigate the expression changes of acetylcholinesterase (AChE) related microRNAs derived from peripheral blood mononuclear cells (PBMCs) in patients with stroke. Methods The microRNAs for targeting AChE mRNA were selected via prediction software and previous studies. PBMCs were extracted from venous blood samples of acute ischemic stroke patients (onset<24 h) and healthy controls. The expressions of microRNAs and AChE mRNA were quantified using real-time PCR (RT-PCR). The protein level of AChE was detected by Western blot assay. Results Thepredicted microRNAs included microRNA (miR)-24,-28,-124,-132,-182*,-194 and-484. The expression levels of miR-24,-124,-132 and-194 were significantly elevated in stroke patients compared with those of controls (P<0.05). There were no significant changes in expression levels of miR-28,-182*and-484. Additionally, the relative expression levels of intracellular AChE mRNA and protein decreased significantly in stroke patients (P<0.05). Conclusion MiRNAs can enhance cholinergic anti-inflammatory pathway by targeting AChE in patients with acute ischemic stroke.