[摘 要] 目的：基于TCGA数据库分析Deltex E3泛素连接酶2（DTX2）在肾透明细胞癌（ccRCC）组织中的表达水平及临床意义，探讨DTX2对ccRCC细胞增殖、迁移和侵袭的影响。方法：利用TIMER数据库分析DTX2在泛癌组织中的表达水平，通过UALCAN数据库进一步验证ccRCC组织和癌旁组织中DTX2 mRNA和蛋白表达差异。使用UALCAN数据库中的TCGA-ccRCC队列数据集，分析ccRCC中DTX2表达与患者临床病理特征的相关性。通过K-M plot数据库分析DTX2表达与ccRCC患者预后的相关性。利用DAVID数据库对DTX2相关基因进行GO和KEGG通路富集分析。通过qPCR法检测DTX2基因在人胚肾293（HEK293）细胞和ccRCC细胞A498、Caki-1中的表达水平。利用siRNA技术分别将DTX2 siRNA及其阴性对照质粒转入A498、Caki-1细胞，采用CCK-8法、平板克隆实验、划痕实验及Transwell侵袭实验分别检测敲低DTX2对细胞增殖、迁移和侵袭的影响。结果：TCGA数据库分析结果表明，与癌旁组织相比，ccRCC组织中DTX2 mRNA和蛋白均呈高表达（均P<0.01）。DTX2表达水平与ccRCC患者的病理分期、临床分级、不同亚型和淋巴结转移相关联（均P<0.01），DTX2高表达与患者的不良预后具有相关性（均P<0.01）。GO功能和KEGG通路富集分析结果显示，DTX2表达相关基因主要参与蛋白酶体介导的泛素依赖性蛋白质分解代谢等生物学过程，并主要富集到了mTOR信号通路等与肿瘤的相关信号通路中（均P<0.05）。体外细胞实验结果表明，A498和Caki-1细胞中DTX2表达水平高于HEK293细胞；敲低DTX2表达可显著降低A498和Caki-1细胞的增殖、迁移及侵袭能力（均P<0.01）。结论：DTX2在ccRCC组织和细胞中呈高表达，其高表达患者的预后较差。敲低DTX2表达可抑制ccRCC细胞增殖、迁移和侵袭，DTX2有望成为ccRCC新的生物标志物和治疗靶点。

Acute B-lymphoblastic leukemia (B-ALL) is the most common type of acute lymphoblastic leukemia. Chemotherapy is the main treatment for most patients, but there are serious side effects. CD19 is generally highly expressed in B-lineage malignancies and plays a key role in B cell signal transduction, activation and development. Therefore, it has become one of the effective targets for treatment of B-cell malignancies. At present, a variety of therapies targeting CD19 have been developed, including antibody drug conjugates, monoclonal antibodies, bispecific antibodies and chimeric antigen receptors cell therapies. This paper reviews the clinical trial progress of CD19-targeted therapy for B-ALL.

Objective To establish a fluorescent assay for rapid detection of Plasmodium falciparum based on recombinaseaided amplification (RAA) and CRISPR-Cas12a system,and to preliminarily evaluate the diagnostic efficiency of this system.. Methods The 18S ribosomal RNA (rRNA) gene of P. falciparum was selected as the target sequence, and three pairs of RAA primers and CRISPR-derived RNA (crRNA) were designed and synthesized. The optimal combination of RAA primers and crRNA was screened and the reaction conditions of the system were optimized to create a fluorescent RAA/CRISPR-Cas12a system. The plasmid containing 18S rRNA gene of the P. falciparum strain 3D7 was generated, and diluted into concentrations of 1 000, 100, 10, 1 copy/μL for the fluorescent RAA/CRISPR-Cas12a assay, and its sensitivity was evaluated. The genomic DNA from P. vivax, P. malariae, P. ovum, hepatitis B virus, human immunodeficiency virus and Treponema pallidum was employed as templates for the fluorescent RAA/CRISPR-Cas12a assay, and its specificity was evaluated. Fifty malaria clinical samples were subjected to the fluorescent RAA/CRISPR-Cas12a assay and nested PCR assay, and the consistency between two assays was compared. In addition, P. falciparum strain 3D7 was cultured in vitro. Then, the culture was diluted into blood samples with parasite densities of 1 000, 500, 200, 50, 10 parasites/μL with healthy volunteers’ O-positive red blood cells for the RAA/CRISPR-Cas12a assay, and the detection efficiency was tested. Results The Pf-F3/Pf-R3/crRNA2 combination, 2.5 μL as the addition amount of B buffer, 40 min as the RAA reaction time, 37 °C as the reaction temperature of the CRISPR-Cas12a system were employed to establish the fluorescent RAA/CRISPR-Cas12a system. Such a system was effective to detect the plasmid containing 18S rRNA gene of the P. falciparum strain 3D7 at a concentration of 1 copy/μL, and presented fluorescent signals for detection of P. falciparum, but failed to detect P. ovum, P. malariae, P. vivax, T. pallidum, hepatitis B virus or human immunodeficiency virus. The fluorescent RAA/CRISPR-Cas12a system and nested PCR assay showed completely consistent results for detection of 50 malaria clinical samples (kappa = 1.0, P < 0.001). Following 6-day in vitro culture of the P. falciparum strain 3D7, 10 mL cultures were generated and the fluorescent RAA/CRISPR-Cas12a system showed the minimal detection limit of 50 parasites/μL. Conclusion The fluorescent RAA/CRISPR-Cas12a system is rapid, sensitive and specific for detection of P. falciparum, which shows promising value for rapid detection and risk monitoring of P. falciparum.

Objective To investigate the prevalence of single nucleotide polymorphisms (SNPs) of artemisinin resistance-related Pfubp1 and Pfap2mu genes in Plasmodium falciparum isolates from Bioko Island, Equatorial Guinea, so as to to provide baseline data for the formulation of malaria control strategies in Bioko Island. Methods A total of 184 clinical blood samples were collected from patients with P. falciparum malaria in Bioko Island, Equatorial Guinea from 2018 to 2020, and genomic DNA was extracted. The Pfubp1 and Pfap2mu gene SNPs of P. falciparum were determined using a nested PCR assay and Sanger sequencing, and the gene sequences were aligned. Results There were 159 wild-type P. falciparum isolates (88.83%) from Bioko Island, Equatorial Guinea, and 6 SNPs were identified in 20 Pfubp1-mutant P. falciparum isolates (11.17%), in which 4 non-synonymous mutations were detected, including E1516G, K1520E, D1525E, E1528D. There was only one Pfubp1gene mutation site in 19 Pfubp1-mutant P. falciparum isolates (95.00%), in which non-synonymous mutations accounted for 68.42% (13/19). D1525E and E1528D were identified as major known epidemic mutation sites in the Pfubp1 gene associated with resistance to artemisinin-based combination therapies (ACTs). At amino acid position 1525, there were 178 wild-type P. falciparum isolates (99.44%) and 1 mutant isolate (0.56%), with such a mutation site identified in blood samples in 2018, and at amino acid position 1528, there were 167 wild-type P. falciparum isolates (93.30%) and 12 mutant isolates (6.70%). The proportions of wild-type P. falciparum isolates were 95.72% (134/140), 79.25% (126/159) and 95.83% (161/168) in the target amplification fragments of the three regions in the Pfap2mu gene (Pfap2mu-inner1, Pfap2mu-inner2, Pfap2mu-inner3), respectively. There were 16 different SNPs identified in all successfully sequenced P. falciparum isolates, in which 7 non-synonymous mutations were detected, including S160N, K199T, A475V, S508G, I511M, L595F, and Y603H. There were 7 out of 43 Pfap2mu-mutant P. falciparum isolates (16.28%) that harbored only one gene mutation site, in which non-synonymous mutations accounted for 28.57% (2/7). For the known delayed clearance locus S160N associated with ACTs, there were 143 wild-type (89.94%) and 16 Pfap2mu-mutant P. falciparum isolates (10.06%). Conclusions Both Pfubp1 and Pfap2mu gene mutations were detected in P. falciparum isolates from Bioko Island, Equatorial Guinea from 2018 to 2020, with a low prevalence rate of Pfubp1 gene mutation and a high prevalence rate of Pfap2mu gene mutation. In addition, new mutation sites were identified in the Pfubp1 (E1504E and K1520E) and Pfap2mu genes (A475V and S508G).

OBJECTIVE: To assess the association of SLC6A4 gene c.*670T>G polymorphism with the risk for asthma and peripheral blood cytological characteristics among ethnic Zhuang Chinese from Guangxi, China. METHODS: From May 2017 to March 2020, 258 patients diagnosed with asthma and 244 healthy controls were recruited from the Affiliated Hospital of Youjiang Minzhu Medical College and the People's Hospital of Hechi. Genotypes of the c.*670T>G polymorphism were determined by Sanger sequencing. Flow cytometry was used in combination with an electrical impedance method for the counting and classification of peripheral blood cells. RESULTS: Compared with the T allele, the G allele of the c.*670T>G polymorphism was associated with the risk for asthma in the population (OR = 1.54, 95%CI = 1.15-2.06; P = 0.004). Compared with the GT and TT genotypes, homozygous GG genotype also comprised a risk factor (OR = 1.66, 95%CI = 1.16-2.38; P = 0.005). Stratification of the risk factors showed that the homozygous GG genotype has increased the risk of asthma in males and urban residents (P < 0.01). The erythrocyte, hemoglobin and platelet counts of the asthma group were significantly higher than the control group (P < 0.001). The GG, GT and TT genotypes have respectively accounted for 82.35%, 17.65% and 0% of the samples with platelets exceeding the normal value. The overall platelet level of GG genotype was higher than GT+TT genotype (P < 0.05). The significant association was verified by the false positive report probability, and at a prior probability level of 0.1, G vs. T false positive probability was 0.071, and GG vs. GT+TT false positive probability was 0.153. CONCLUSION The GG genotype of the c.*670T>G polymorphism is associated with the risk for asthma among ethnic Zhuang Chinese from northwest Guangxi. Above finding has also enriched the genotypic data and peripheral blood phenotype for this polymorphism.
Male ; Humans ; East Asian People ; China ; Genotype ; Alleles ; Asthma/genetics* ; Serotonin Plasma Membrane Transport Proteins

OBJECTIVE: To compare clinical efficacy of No-touch technique and traditional retractor in treating calcaneal fracture. METHODS: Clinical data of 74 calcaneal fracture patients with closed Sanders typeⅡ to Ⅳ were retrospectively analyzed from July 2019 to June 2021. According to different treatment methods, the patients were divided into No-touch group and conventional group, 37 patinets in each group. In No-touch group, there were 25 males and 12 females, aged from 19 to 70 years old with an average of (42.64±14.16) years old;17 patients were typeⅡ, 14 patinets with type Ⅲ, 6 patients with type Ⅳ according to Sanders fracture classification;three 2.0 mm Kirschner wires were implanted into the talus body, talus neck, and cuboid bone, and the flap was turned upward to expose the operation area. In conventional group, there were 30 males and 7 females, aged from 19 to 67 years old with an average of (41.56±11.38) years old;17 patients with typeⅡ, 12 patients with type Ⅲ, 8 patients with type Ⅳ according to Sanders fracture classification;the operation was completed by exposing the operation area with traditional retractor. Operation time, postoperative incision complications, postoperaive American Orthopedic Foot and Ankle Society (AOFAS) ankle hind foot score at 6 months between two groups were compared. RESULTS: Seventy-four patients were followed up, and follow-up time in No-touch group ranged from 6 to 17 months with an average of(9.57±2.72) months, while in conventional group ranged from 6 to 16 months with an averge of(9.14±2.71) months, and no difference in follow-up between two groups (P>0.05). Operation time in No-touch group (55.67±7.94) min was shorter than that in conventional group (70.16±9.41) min (P<0.05);four patients in No-touch group occurred incision complications, while 8 patients in normal group, and had statistically difference(P<0.05). Daily activities and support, maximum walking distance (block), ground walking, limited degree of flexion, extension and valgus, foot alignment and total score of AOFAS scores in No-touch group was significantly higher than that of conventional group (P<0.05). There were no significant difference in pain degree, abnormal gait and ankle hind foot stability between two groups(P>0.05). According to AOFAS score, 19 patients got excellent result, 16 good and 2 poor in No-touch group;while 9 excellent, 24 good, and 4 poor in conventional group, and no difference between two groups (P>0.05). CONCLUSION Compared with traditional retractor in treating calcaneal fracture, No-touch technology could significantly shorten operation time, reduce incidence of postopertive complications, while two methods could improve excellent and good rate of ankle joint function recovery after operation.
Male ; Female ; Humans ; Young Adult ; Adult ; Middle Aged ; Aged ; Fracture Fixation, Internal ; Retrospective Studies ; Calcaneus/surgery* ; Fractures, Bone/surgery* ; Ankle Injuries ; Treatment Outcome ; Ankle Joint ; Foot Injuries ; Knee Injuries ; Postoperative Complications ; Talus

[Abstract] Objective To explore the distribution situation of microRNA(miR) -30 gene single-nucleotide sites rs1192037A / T polymorphisms in Guangxi Zhuang population and compare its distribution differences with other populations and to analyze level of common blood lipid indexes in genotypes. Methods SNPscan was used to detect rs1192037A / T locus genotyping in 236 volunteers of Zhuang nationality in Guangxi. The genotypes and allele frequencies of rs1192037A / T locus genotyping in different genders and groups were analyzed. The levels of common blood lipids in the subjects were detected by roche automatic biochemical apparatus. Results Three genotypes of AA, AT and TT were found in rs1192037 A / T with the frequency distribution of 11. 0%, 38. 6% and 50. 4%, respectively. No significant differences in genotypes and alleles frequencies of rs1192037 A / T between different genders in Guangxi Zhuang population were observed (P > 0. 05) . However,there were significant differences in the genotype and allele frequency of miR-30 gene rs1192037 A / T in Guangxi Zhuang population compared with those of Europeans, Japanese, Africans, Mexicans and Indians published by HapMap (P<0. 05), and there were not significant difference in genotypes and allele frequencies in population of HCB and JTP (P>0. 05) . There were significant differences in the levels of TG among the 3 genotypes of rs1192037 A / T, and the TG levels of AT and TT genotypes were significantly higher than AA genotypes. Conclusion There are different degrees of rs1192037 A / T polymorphisms of miR-30 gene among Guangxi population and other ethnic populations and other regions. The polymorphism of rs1192037 A / T is related to the level of TG.

【Objective】 To explore the distribution of polymorphisms of miR-208 genes rs8022522 and rs12894524 locus in Guangxi healthy population and compare the differences in the polymorphism distribution in different population. 【Methods】 SNPscan technology was used to detect genotypes of rs8022522 and rs12894524 from 297 healthy people in Guangxi, and the results were compared with other populations from Human genome Haplotype Map(HapMap) data. 【Results】 Three genotypes, namely, AA (2.7%), AG (24.2%) and GG (73.1%), in rs8022522 were found, with the allele frequencies of A and G being 14.8% and 85.2%. The genotypes of rs12894524 locus were TT (1.3%), TG (13.5%) and GG (85.2%), and the frequency of T and G allele was 8.1% and 91.9%, respectively. rs8022522 and rs12894524 locus genotypes and allele frequencies were significantly different from HapMap-CEU, HapMap- YRI and HapMap-TSI (P<0.05). Compared with HapMap-JPT and HapMap-CHB, there was no significant difference in genotype or allele frequency between the two sites (P>0.05). As for the blood lipid level among the three genotypes in rs8022522, the level of high density lipoprotein cholesterol (HDL-C) with GG genotype was significantly different from that in AG group (P<0.05). 【Conclusion】 The polymorphisms of rs8022522 and rs12894524 of miR-208 gene in Guangxi population are different from those in other regions to varying degrees. The polymorphism of rs8022522 locus is related to the level of HDL-C.

Connexin (Cx), a multigene-encoded transmembrane protein family, forms either gap junctions ( GJ) or hemichannels (HC) to mediate intercellular communication in plasma mem¬brane between adjacent cells or interacts with proteins by its car- boxyl terminal in the cytoplasm to participate in the process of tumor cell proliferation, apoptosis, necrosis, invasion, metasta¬sis, drug resistance and stem cell characteristics.However, mi- slocalization of Cx in cytoplasm or nucleus often occurs in many tumors, and involved in the occurrence and development of tumors.Subcellular localization of Cx is affected by post-transla- tional modifications, including phosphorylation, ubiquitination, and acetylation.In this paper the classification and function of Cx, the relationship between subcellular localization of Cx and tumorigenesis and the regulation of post-translational modifica¬tion on Cx are reviewed in order to provide new ideas for the study of Cx as a potential target for cancer therapy.

Objective To investigate the genetic polymorphisms of Plasmodium falciparum multidrug resistance protein 1 (PfMDR1), chloroquine resistance transporter (PfCRT) and Kelch 13 (PfK13) genes in Bioko Island, Equatorial Guinea, so as to provide insights into the development of the malaria control strategy in local areas. Methods A total of 85 peripheral blood samples were collected from patients with Plasmodium falciparum infections in Bioko Island, Equatorial Guinea in 2018 and 2019, and genomic DNA was extracted. The PfMDR1, PfCRT and PfK13 genes were amplified using a nested PCR assay. The amplification products were sequenced, and the gene sequences were aligned. Results There were no mutations associated with artemisinin resistance in PfK13 gene in Bioko Island, Equatorial Guinea, while drug-resistant mutations were detected in PfMDR1 and PfCRT genes, and the proportions of PfMDR1_N86Y, PfMDR1_Y184F and PfCRT_K76T mutations were 35.29% (30/85), 72.94% (62/85) and 24.71% (21/85), respectively. Conclusion There are mutations in PfMDR1, PfCRT and PfK13 genes in P. falciparum isolates from Bioko Island, Equatorial Guinea.