Prokaryotic expression and immune function of Mycobacterium tuberculosis Rv3133 c gene / 中华微生物学和免疫学杂志

Objective:To construct a plasmid for expression Mycobacterium tuberculosis ( Mtb) Rv3133c and to evaluate the immunogenicity of Rv3133c through population and mice experiments. Methods:The recombinant expression plasmid pPROEX-Rv3133c was constructed. The transformed E. coli BL21 (DE3) carrying expression plasmid was induced by IPTG to express the recombinant Rv3133c (rRv3133c). Western blot was used to identify the expressed protein. Whole-blood IFN-γ release assay (WBIA) was preformed to assess the immunogenicity of rRv3133c in Mtb-infected population. Antigen-specific antibodies in serum, Th1 type cytokines in splenocytes, functional T cell subset responses in splenocytes and the expression of cytokines at mRNA level in lung tissues were detected after immunizing mice subcutaneously with rRv3133c and adjuvant DC. Results:The rRv3133c was constructed and expressed successfully. Stimulation with rRv3133c promoted the production of IFN-γ in Mtb-infected population, especially in patients with latent tuberculosis infections. After immunizing mice with rRv3133c+ DC, the levels of IFN-γ, TNF-α and IL-2, the number of IFN-γ + TNF-α + CD4 + T cells in spleen and the expression of antigen-specific IFN-γ, TNF-α and iNOS at mRNA level in lung tissues were higher than those in BCG-immunized mice, but lower than those in BCG+ rRv3133c+ DC group. The serum IgG2a/IgG1 ratios in the rRv3133c+ DC group and the BCG+ rRv3133c+ DC group were greater than 1, and significantly higher than that of the BCG group. Conclusions:The rRv3133c had good immunogenicity and could induce strong Th1 immune response, suggesting that it was a potential candidate antigen for subunit vaccine against tuberculosis.

Clinical characteristics of 272 437 patients with different histopathological subtypes of primary esophageal malignant tumors / 中华内科杂志

Objective:To characterize the histopathological subtypes and their clinicopathological parameters of gender and onset age by common, rare and sparse primary esophageal malignant tumors (PEMT).Methods:A total of 272 437 patients with PEMT were enrolled in this study, and all of the patients were received radical surgery. The clinicopathological information of the patients was obtained from the database established by the State Key Laboratory of Esophageal Cancer Prevention & Treatment from September 1973 to December 2020, which included the clinical treatment, pathological diagnosis and follow-up information of esophagus and gastric cardia cancers. All patients were diagnosed and classified by the criteria of esophageal tumor histopathological diagnosis and classification (2019) of the World Health Organization (WHO). The esophageal tumors, which were not included in the WHO classification, were analyzed separately according to the postoperative pathological diagnosis. The χ 2 test was performed by the SPSS 25.0 software on count data, and the test standard α=0.05. Results:A total of 32 histopathological types were identified in the enrolled PEMT patients, of which 10 subtypes were not included in the WHO classification. According to the frequency, PEMT were divided into common (esophageal squamous cell carcinoma, ESCC, accounting for 97.1%), rare (esophageal adenocarcinoma, EAC, accounting for 2.3%) and sparse (mainly esophageal small cell carcinoma, malignant melanoma, etc., accounting for 0.6%). All the common, rare, and sparse types occurred predominantly in male patients, and the gender difference of rare type was most significant (EAC, male∶ female, 2.67∶1), followed with common type (ESCC, male∶ female, 1.78∶1) and sparse type (male∶ female, 1.71∶1). The common type (ESCC) mainly occurred in the middle thoracic segment (65.2%), while the rare type (EAC) mainly occurred in the lower thoracic segment (56.8%). Among the sparse type, malignant melanoma and malignant fibrous histiocytoma were both predominantly located in the lower thoracic segment (51.7%, 66.7%), and the others were mainly in the middle thoracic segment.Conclusion:ESCC is the most common type among the 32 histopathological types of PEMT, followed by EAC as the rare type, and esophageal small cell carcinoma and malignant melanoma as the major sparse type, and all of which are mainly occur in male patients. The common type of ESCC mainly occur in the middle thoracic segment, while the rare type of EAC mainly in the lower thoracic segment. The mainly sparse type of malignant melanoma and malignant fibrous histiocytoma predominately occur in the lower thoracic segment, and the remaining sparse types mainly occur in the middle thoracic segment.

Exploration of the post competency-based teaching mode of English for medical laboratory / 中华医学教育探索杂志

English for medical laboratory is an instrumental course,which aims to develop students'competency in reading English literature,writing English scientific papers and conducting international academic exchanges in English.With the continuous expansion and deepening of international exchange and cooperation in laboratory medicine and the demands for training international talents in higher education in China,higher requirements have been put forward for the teaching quality of this course.According to the post competency requirements for professionals of medical laboratory,this paper mainly discusses the teaching mode of English for medical laboratory from the aspectsof educational concept,teaching material construction,teaching mode,assessment mechanism and teachers,hoping to play an active role in the teaching reform of English for medical laboratory at the undergraduate level.

Identification of a novel mutation of COL2A1 gene in a Chinese family affected with spondyloepiphyseal dysplasia congenita / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To identify potential disease-causing mutation in the COL2A1 gene in a Chinese family affected with autosomal dominant spondyloepiphyseal dysplasia congenita (SEDC; OMIM 183900) and to analyze the phenotype-genotype correlation.</p><p><b>METHODS</b>Complete physical, and radiographic examinations of 4 affected individuals from the family were conducted. Genomic DNA was isolated from peripheral blood leukocytes. Whole-exome sequencing was performed using a HiSeq2000 sequencer. All 54 exons and exon-intron boundaries of the COL2A1 gene were amplified by polymerase chain reaction (PCR) and bidirectionally sequenced.</p><p><b>RESULTS</b>All of the 4 individuals were found to carry a novel missense mutation of c.2224G>A (p.Gly687Ser) in the COL2A1 gene, while the same mutation was not found in the normal members of the family and 50 healthy controls. Protein prediction of missense mutation by Polyphen-2 and SIFT software indicated severe damage to the function.</p><p><b>CONCLUSION</b>The mutation c.2224G>A (p.Gly687Ser) of the COL2A1 gene is responsible for this family. There are heterozygous of phenotype for the mutation.</p>

Effects of brucine on chronic myeloid leukemia cell line HL-60 / 白血病·淋巴瘤

Objective To investigate the proliferation inhibition and the apoptosis induction effect of brucine on human chronic myeloid leukemia cell line HL-60 cells.Methods HL-60 cells were exposed to various dosages of brucine 24,48,72 h respectively,MTT method was used to assay the growth inhibition effect of brucine on HL-60 cells and the IC50 of brucine was evaluated at the same time.The morphology was observed by AO/EB stains.The cell apoptosis and cell cycle was tested by flow cytometry with Annexin V-FITC/PI double staining and PI labeling respectively.Results The results indicated that the brucine displayed significant anti-proliferative effect on HL-60 cells in a dose-and time-dependent manner,with IC50 value of 211.8 μg/ml(24 h),107 μg/ml(48 h),83 μg/ml(72 h)respectively.The most significant inhibition was observed at 320 μg/ml for 48 h.In this condition,apoptosis morphology was induced by brucine with nuclear chromatin condensation,most of the nuclears were orange stained and condensation-like or bead-like,which was consistent with the Annexin V-FITC/PI results.The cell apoptosis rates were(2.1±1.1)％,(21.3±1.2)％,(38.6±1.3)％,(58.5±4.1)％,(75.3±0.87)％and(66.2±0.75)％in different dose of brucine,respectively.At the same time,the cell cycle analysis results showed that the cell ratio in G0/G1 phase was decreased while in G2/M and sub-G0 phases was increased,comparing with blank control group.Conclusion Brucine can inhibit cell growth dramatically,which may be related to the cell apoptosis and the cell cycle arrest.

The relationship between C20orf54 gene rs3746804 position single nucleotide polymorphism and susceptibility to esophageal squamous cell carcinoma / 中华内科杂志

Objective To explore the association of C20orf54 gene rs3746804 position single nucleotide polymorphism and susceptibility to esophageal squamous cell carcinoma (ESCC).Methods Purification of genomic DNA from whole blood was used the Maxwell(R) 16 System.rs3746804 in C20ort54 was detected by direct sequencing in 434 ESCC patients from Changzhi (Shanxi province) and Linzhou (Henan province) and 554 healthy controls from Changzhi,Linzhou and including immigrators from Linzhou to Changzhi.Results For rs3746804,the genotypic frequencies of CT(37.5％ vs 51.0％,37.5％ vs 52.0％),CC (44.2％ vs 34.8％,44.2％ vs 33.0％) in Changzhi ESCC patients showed significant differences with healthy Changzhi controls and the healthy immigrator controls (all P ＜ 0.05),and the frequencies of TT(18.3％ vs 4.1％) and CC (44.2％ vs 54.6％) in Changzhi ESCC patients showed significant differences with Linzhou ESCC patients (all P ＜0.05).The genotypic frequencies of TT (4.1％ vs 15.0％),CT (41.2％ vs 52.0％) and CC(54.6％ vs 33.0％) showed significant differences between Linzhou ESCC patients and the healthy immigrator controls (all P ＜ 0.05),and the frequencies of TT (4.1％ vs 14.1％) and CC (54.6％ vs 34.8％) showed significant differences between Linzhou ESCC patients and Changzhi healthy controls (all P ＜ 0.01).Meanwhile,there were significant differences between ESCC patients (including Changzhi and Linzhou ESCC patients) and healthy controls (including the healthy Changzhi,Linzhou and immigrator controls) in genotypic frequencies of CT(39.2％ vs 48.7％) and CC (48.8％ vs 38.2％) (all P ＜ 0.01).CT and CT + TT genotype could decrease the risk of ESCC compared with the CC genotype (OR =0.630,95％ CI0.481-0.826 ; OR =0.654,95％ CI 0.507-0.844).Conclusion There is a closed relationship between SNP rs3746804 in C20orf54 and susceptibility to ESCC.

A comparison and significance of plasma riboflavin levels in patients with esophageal squamous cell carcinoma versus Linzhou healthy migrants in Changzhi of Shanxi / 中华内科杂志

ObjectiveTo study the relationship between plasma riboflavin levels and esophageal squamous cell carcinoma.Methods We detected and compared plasma concentrations of riboflavin in patients with esophageal squamous cell carcinoma (ESCC) and immigrants of Linzhou living in Changzhi.Plasma riboflavin levels were quantified in 445 ESCC patients,689 healthy control subjects and 347 immigrants of Linzhou living in Changzhi by using enzyme-linked immunosorbent assay.ResultsThe plasma riboflavin levels in patients with ESCC were significantly lower than those in the healthy controls and immigrants of Linzhou living in Changzhi [ (731.69 ± 330.67 ) μg/L vs ( 1090.43 ± 445.08 ) μg/L,(731.69 ± 330.67) μg/L vs ( 897.58 ± 177.78) μg/L,respectively,all P ＜ 0.05 ],and the plasma riboflavin levels of the healthy controls were higher than those in the immigrants of Linzhou living in Changzhi (P ＜ 0.05).ConclusionPatients with ESCC have decreased plasma riboflavin levels as compared with the healthy controls and immigrants of Linzhou living in Changzhi,there exists a lack of riboflavin in ESCC patients,but the specific mechanism needs further study.

A study on the expression of survivin, livin and XIAP mRNA in peripheral blood of patients with gastric cancer / 中华普通外科杂志

Objective To investigate the mRNA expression of survivin, livin and XIAP gene in peripheral blood of patients with gastric cancer and its relationship with clinico-pathological features.Methods This study included 50 patients with gastric cancer and 20 healthy donors. The expression of survivin, livin and XIAP gene was detected by reverse transcription-quantitative polymerase chain reaction (RT-QPCR) using a molecular beacon probe, while recombination plasmid containing the sequence of survivin, livin and XIAP was standard. The relationship between copies of survivin, livin and XIAP gene expression in peripheral blood with gastric cancer and clinical data was analyzed. Results A linear standard curve was obtained between 103 ～ 1010 copies. The copies of survivin, livin and XIAP mRNA in peripheral blood of patients with gastric cancer did not correlate with gender, age, and histological types ( P ＞ 0.05). There were positive relationships between copies of survivin, livin and XIAP gene with lymph node metastasis and TNM stage (P ＜0.05 ). The expression of survivine, livin and XIAP was all negative in peripheral blood of healthy people. 52% (17/33) of patients suffered from recurrence or metastasis who had positive expression of survivin and/or livin and/or XIAP mRNA, while it was 18% (3/17)among the negative survivin and/or livin and/or XIAP mRNA caces ( P ＜ 0.05). Conclusions The expression of survivin, livin and XIAP mRNA can be used to detecte micro-metastasis in peripheral blood circulation of gastric cancer.