A newly discovered Anaplasma phagocytophilum variant in rodents from southeastern China

Anaplasma phagocytophilum was detected by polymerase chain reaction in 13 (14.1%) of 92 rodents captured from a mountainous area of Zhejiang Province in southeastern China. The nucleotide sequences of 1442-bp, nearly entire 16S rRNA gene amplified from these rodents, had 100% identity, but varied from all known corresponding sequences of A. phagocytophilum deposited in GenBank. To further identify and classify the variant, fragments of 357-bp partial citrate synthase gene (gltA), 849-bp major surface protein 4 gene (msp4), and 443-bp groESL heat-shock operon gene, were amplified and analyzed. The nucleotide sequences of the partial gltA gene amplified from the rodents were identical to each other, but distinct from previously reported A. phagocytophilum sequences,as were msp4 and groESL. These findings indicate that the newly discovered agent represents a novel A. phagocytophilum variant.     

茵虎利胆合剂配合鼻胆管引流治疗阻塞性黄疸30例

目的:观察茵虎利胆合剂配合鼻胆管引流术(ENBD)治疗阻塞性黄疸的临床效果,并探讨其作用机制.方法:将90例阻塞性黄疸患者随机分成手术对照组、内镜对照组以及茵虎实验组各30例.手术组行传统开放手术治疗:内镜组行ENBD治疗.实验组给予ENBD+茵虎利胆合剂治疗.观察各组患者T管、鼻胆管引流情况,同时监测90例患者术前、术后7、14 d的血清TBIL、DBIL、AKP、GPT、GOT、LPS的变化和手术前后IL-6水平变化.结果:术后第14天各组患者血清LPS、IL-6、TBIL、DBIL、AKP、GPT、GOT较术前均显著下降(P＜0.05或0.01),且茵虎实验组明显低于手术及内镜对照组(0.07±0.01vs 0.12±0.03,0.13±0.03;166.5±31.62vs 230.2±41.08,225.6±42.69;24.3±7.91vs 43.3±9.49,46.7±11.07;12.6±4.74vs 32.2±6.33,46.7±11.07;67.5±25.30vs 102.5±36.37,114.9±39.53;36.8±14.23vs 79.0±35.88,82.3±34.79;46.2±11.07vs 84.5±34.79,80.3±31.62,P＜0.05或0.01),手术对照组及内镜对照组间无显著性差异;术后5d始茵虎实验组胆汁引流水平明显高于手术及内镜对照组(P＜0.05或0.01),而手术与内镜对照组间无显著性差异.结论:茵虎利胆合剂配合鼻胆管引流可以有效地治疗阻塞性黄疸,促进胆汁排泄.降低LPS和IL-6.     

Expression of p21(WAF1) and p53 and polymorphism of p21(WAF1) gene in gastric carcinoma

AIM: To investigate the relationship between expression of p21(WAF1) and p53 gene, and to evaluate the deletion and polymorphism of p21(WAF1) gene in gastric carcinoma (GC). METHODS: Expression of p21 and p53 proteins, and deletion and polymorphism of p21 gene in GC were examined by streptavidin-peroxidase conjugated method (SP) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) respectively. RESULTS: The expression of p21 and p53 was found in 100% (20/20) and 0% (0/20) of normal gastric mucosae(NGM), 92.5% (37/40) and 15.0% (6/40) of dysplasia (DP) and 39.8% (43/108) and 56.5% (61/108) of GC, respectively. The positive rate of p21 in GC was lower than that in NGM and DP (P<0.05), while the positive rate of p53 in GC was higher than that in NGM and DP (P<0.05). p21 and p53 were significantly expressed in 63.3% (19/30) and 36.7% (11/30), 35.0% (14/40) and 77.5% (31/40), 26.7% (4/15) and 80.0% (12/15), 30.8% (4/13) and 30.8% (4/13), and 20.0% (2/10) and 30.0% (3/10) of well-differentiated, poorly-differentiated, undifferentiated carcinomas, mucoid carcinomas and signet ring cell carcinomas. The expression of p21 in well-differentiated carcinomas was significantly higher than that in poorly-differentiated, un-differentiated, mucoid carcinomas and signet ring cell carcinomas (P<0.05). Contrarily, The expression of p53 was increased from well-differentiated to poorly-differentiated and un-differentiated carcinomas (P<0.05). The expression of p21 and p53 in paired primary and metastatic GC (35.3% and 70.6%) was different from non-metastatic GC (62.5% and 42.5%) markedly (P<0.05). The expression of p21 in invasive superficial muscle (60.0%) was higher than that in invasive deep muscle or total layer (35.2%) (P<0.05) and was higher in TNM stages I (60.0%) and II (56.2%) than in stages III (27.9%) and IV (22.2%) (P<0.05), whereas the expression of p53 did not correlate to invasion depth or TNM staging (P>0.05). The exoression patterns of p53+/p21-, and of p53-/p21+ were found in 5.0% and 82.5% of DP. There was a significant correlation between expression of p21 and p53 (P<0.05). But there was no significant correlation between expression of both in GC (P>0.05). There was no deletion in exon 2 of p21 gene in 30 cases of GC and 45 cases of non-GC, but polymorphism of p21 gene at exon 2 was found in 26.7% (8/30) of GC and 8.9% (4/45) of non-GC, a significant difference was found between GC and non-GC (P<0.05). There was no significant relation between p21 expression of polymorphism (37.5%, 3/8) and non-polymorphism (45.5%, 10/22) in GC (P>0.05). CONCLUSION: The loss of p21 protein and abnormal expression of p53 are related to carcinogenesis, differentiation and metastasis of GC. The expression of p21 is related to invasion and clinical staging in GC intimately. The expression of p21 protein depends on p53 protein largely in NGM and DP, but not in GC. No deletion of p21 gene in exon 2 can be found in GC. The polymorphism of p21 gene might be involved in gastric carcinogenesis.There is no significant association between polymorphism of p21 gene and expression of p21 protein.     

鱼胆中毒及其并发多器官功能障碍综合征患者血浆内皮素水平的研究

目的探讨血浆内皮素(ET)在鱼胆中毒患者的变化及其与鱼胆中毒后多器官功能障碍综合征(MODS)发生及发展的关系.方法采用放射免疫法测定85例鱼胆中毒患者与30名健康人血浆ET水平,比较鱼胆中毒各组间血浆ET水平,分析鱼胆中毒患者血浆ET水平与血清酶学、血生化等指标的关系.结果鱼胆中毒患者血浆ET水平(150.5±90.5) ng/L较正常对照组(52.8±9.3) ng/L明显升高(P<0.01),轻度中毒组血浆ET水平(61.2±13.6) ng/L与正常对照组比较,差异无显著性(P>0.05),中度与重度中毒组血浆ET水平(162.3±82.3) ng/L,(232.6±92.7) ng/L较正常对照组明显升高(P值均<0.01),中度中毒组血浆ET水平高于轻度中毒组(P<0.01),重度中毒组又高于中度中毒组(P<0.01).MODS组与非MODS组血浆ET水平(210.2±98.7) ng/L,(126.3±68.3) ng/L均显著高于对照组(P<0.01),MODS组血浆ET水平又高于非MODS组(P<0.01).血浆ET与肌酸激酶同工酶、心肌肌钙蛋白I 、乳酸脱氢酶、ALT、AST、血尿素氮、血肌酐呈显著正相关(P<0.01或0.05),与肌酐清除率呈显著负相关(P<0.01).死亡组血浆ET水平(253.5±96.7) ng/L明显高于非死亡组(159.3±67.9) ng/L(P<0.01).结论 ET参与了鱼胆中毒的发病及其中毒后MODS的发生与进展.鱼胆中毒患者中毒程度越重,血浆ET升高就越明显,病死率也就越高.动态监测血浆ET的水平可作为判断鱼胆中毒患者病情轻重、预后的一个重要临床指标,对早期判断或警示鱼胆中毒后MODS的发生,并早期应用ET拮抗剂或抑制剂防治鱼胆中毒后MODS的发生发展、降低病死率有重要的临床价值.     

Polygalasaponin F inhibits neuronal apoptosis induced by oxygen‐glucose deprivation and reoxygenation through the PI3K/Akt pathway

Cerebral ischaemia is a common cerebrovascular disease and often induces neuronal apoptosis, leading to brain damage. Polygalasaponin F (PGSF) is one of the components in Polygala japonica Houtt, and it is a triterpenoid saponin monomer. This research focused on anti‐apoptotic effect of PGSF during oxygen‐glucose deprivation and reoxygenation (OGD/R) injury in rat adrenal pheochromocytoma cells (PC12) and primary rat cortical neurons. OGD/R treatment reduced viability of PC12 cells and primary neurons. This reduced viability was prevented by PGSF, as shown by MTT assay. OGD/R insult decreased expression of Bcl‐2/Bax both in PC12 cells and primary neurons but elevated levels of caspase‐3 in primary neurons. However, PGSF may up‐regulate expression of Bcl‐2/Bax and down‐regulate caspase‐3 in these particular cells. Furthermore, Bcl‐2/Bax and the ratio between phosphorylated Akt and total Akt were decreased in PC12 cells treated with OGD/R, and both were increased by PGSF. Moreover, increase in the ratios of Bcl‐2/Bax and phosphorylated Akt/total Akt in PC12 cells was suppressed by phosphatidylinositol 3‐kinase (PI3K) inhibitor. Data suggest PGSF might prevent OGD/R‐induced injury via activation of PI3K/Akt signalling. The ability of PGSF to block the effects of OGD/R appears to involve regulation of Bcl‐2, Bax and caspase‐3, which are related to apoptosis.     

A combination of ssGSEA and mass cytometry identifies immune microenvironment in muscle‐invasive bladder cancer

BackgroundMuscle‐invasive bladder cancer (MIBC) is a heterogeneous disease with varying clinical courses and responses to treatment. To improve the prognosis of patients, it is necessary to understand such heterogeneity.MethodsWe used single‐sample gene set enrichment analysis to classify 35 MIBC cases into immunity‐high and immunity‐low groups. Bioinformatics analyses were conducted to compare the differences between these groups. Eventually, single‐cell mass cytometry (CyTOF) was used to compare the characteristics of the immune microenvironment between the patients in the two groups.ResultsCompared with patients in the immunity‐low group, patients in the immunity‐high group had a higher number of tumor‐infiltrating immune cells and greater enrichment of gene sets associated with antitumor immune activity. Furthermore, positive immune response‐related pathways were more enriched in the immunity‐high group. We identified 26 immune cell subsets, including cytotoxic T cells (Tcs), helper T cells (Ths), regulatory T cells (Tregs), B cells, macrophages, natural killer (NK) cells, and dendritic cells (DCs) using CyTOF. Furthermore, there was a higher proportion of CD45+ lymphocytes and enrichment of one Tc subset in the immunity‐high group. Additionally, M2 macrophages were highly enriched in the immunity‐low group. Finally, there was higher expression of PD‐1 and Tim‐3 on Tregs as well as a higher proportion of PD‐1+ Tregs in the immunity‐low group than in the immunity‐high group.ConclusionIn summary, the immune microenvironments of the immunity‐high and immunity‐low groups of patients with MIBC are heterogeneous. Specifically, immune suppression was observed in the immune microenvironment of the patients in the immunity‐low group.     

Thromboelastography (TEG) in normal pregnancy and its diagnostic efficacy in patients with gestational hypertension,gestational diabetes mellitus,or preeclampsia

BackgroundThromboelastography (TEG) provides global assessment of hemostatic function and has been recommended to monitor potential coagulopathies during pregnancy in which hypercoagulable state is favored. In present study, we established the reference intervals (RIs) of the TEG parameters (R, K, MA, and α‐angle) with Chinese pregnant women of third trimester. In addition, we examined the diagnostic efficacies of the TEG parameters in the patients diagnosed of gestational hypertension (GH), gestational diabetes mellitus (GDM), or preeclampsia (PE).MethodsWith specified including and excluding criteria, non‐pregnant controls, healthy pregnant women, and pregnant women with GH, GDM, or PE had their venous blood drawn at Beijing Obstetrics and Gynecology Hospital, followed by TEG tests performed in the clinical laboratory.ResultsThe RIs determined with the healthy pregnant women (in third trimester) for R, K, MA, and α‐angle were 4.0‐7.7, 1.2‐3.2, 51.9‐70.1, and 41.4‐74.4, respectively. When compared with the healthy pregnancy group, the K value was significantly decreased in GH patients but increased in PE patients; MA was significantly lower in the PE group. In the receiver operating characteristic curve (ROC) analyses, K value was able to efficiently distinguish normal pregnancy from the GH patients, with an AUC of 0.86 which is far better than those of R (AUC = 0.57) and MA (AUC = 0.56). For the PE patients, the AUC of MA (0.69) was significantly greater than that of R (0.50).ConclusionsThromboelastography may provide more accurate experimental basis for monitoring coagulation functions especially in pregnant women with complications of GH and PE.     

USP21 promotes cell proliferation by maintaining the EZH2 level in diffuse large B‐cell lymphoma

BackgroundDiffuse large B‐cell lymphoma (DLBCL) is the most common category of non‐Hodgkin lymphoma (NHL). However, the underlying molecular mechanism of DLBCL remains unclear.MethodsReal‐time PCR and Western blot analysis were performed to assess the expression of ubiquitin‐specific peptidase 21 (USP21) or enhancer of zeste 2 polycomb repressive complex 2 subunit (EZH2). CCK8 assay and cell death staining were carried out to examine the role of USP21 in cell proliferation and cell death, respectively.ResultsWe found that the deubiquitinase USP21 was highly expressed in the DLBCL lymphoid tissue. The expression of USP21 promoted DLBCL cell proliferation, while it had no obvious effect on cell death. In addition, we found that USP21 regulated cell proliferation via cysteine 221, the catalytic site of USP21. Furthermore, we identified that USP21 could stabilize EZH2, a protein required for germinal center formation and lymphoma formation.ConclusionThe deubiquitinase USP21 promotes cell proliferation by maintaining the EZH2 protein level in DLBCL.     

Establishing reference intervals for vitamins A and E in Chinese elderly people using liquid chromatography‐tandem mass spectrometry

BackgroundVitamins A and E play important roles in sustaining life activities and maintaining a good physical condition. However, most people, particularly the elderly, experience micronutrient deficiencies. This study aimed to establish reference intervals (RIs) for vitamins A and E in Chinese elderly people using a liquid chromatography‐tandem mass spectrometry (LC‐MS/MS) method.MethodsA total of 356 apparently healthy individuals aged ≥64 years who underwent health checkups were randomly selected for the study. Vitamin A and E levels were measured using LC‐MS/MS. The effect of sex on vitamin A and E levels was evaluated, and RIs were established using a parametric method.ResultsFemales showed significantly higher levels of vitamin E than males (p < 0.05). However, no significant sex‐specific difference was observed with vitamin A levels. The RI for vitamin A in the elderly was 0.283–0.730 mg/L. For vitamin E, the RIs were 4.39–15.63, 4.51–16.14, and 4.41–14.67 mg/L for the total, female, and male participants, respectively. In multiple linear regression, alanine aminotransferase, glutamyl transpeptidase, urea, glucose, and uric acid levels increased with increasing vitamin A levels (p < 0.05), and total cholesterol and low‐density lipoprotein cholesterol levels increased with increasing vitamin E levels (p < 0.05). Direct bilirubin levels decreased with increasing vitamin E levels (p < 0.05).ConclusionsThis study established RIs for vitamins A and E in Chinese elderly individuals using an LC‐MS/MS method. We also found that females had significantly higher vitamin E levels than males. The findings could provide a scientific basis for interpreting vitamin status in the elderly.