1.Analysis of ABO serological phenotype and molecular biology results of 256 blood donors with ambiguous blood group
Lanlan CHEN ; Yanhua ZHANG ; Jin CHE ; Feng LI ; Jingmin MA ; Jiahong ZHANG
Chinese Journal of Blood Transfusion 2024;37(7):766-772
Objective To correctly identify the blood group of ABO and study its molecular biological characteristics.Methods Blood samples from blood donors with discrepancies in forward and reverse typing using the microplate method were subjected to both saline tube agglutination test for serological identification and polymerase chain reaction-sequence specific primers(PCR-SSP)for genotyping.Additionally,direct sequencing of exons 1 to 7 of the ABO gene was performed on some donor samples to analyze their blood phenotype,genotype and gene sequence.Results For 256 samples showing discrepancy between forward and reverse typing in microwell method,119 were identified as normal ABO blood types,90 were weakened ABO antibodies and 47 were ABO subtypes by serology tube test.According to the PCR-SSP genotyping test,233 of 256(91.02%)were consistent with serological phenotype and genotype,17 of 256(6.64%)were inconsistent,and 6 samples can′t read genotype based on the kit result typing table.A total of 17 genotypes were identified in 250 samples as AO1 in 56,AO2 in 58,AA in 50,BO1 in 31,BO2 in 17,BB in 8,O1O1 in 2,O1O2 in 7,AB in 13,AO4 in 1,A205O2 in 1,A205A in 1,A201A in 1,O1O4 in 1,O2O2 in 1,A201B in 1 and A205B in 1.Sequencing of exons 1 to 7 of the ABO gene was carried out in 78 samples,and 29 ABO alleles were detected,seven of which were common alleles(?A101,?A102,?A104,?B101,?O01,?O02,?O04),22 of which were rare alleles(?A201,?A205,?Ax01,?Ax03,?Ax13,?Ax19,?Ax22,?Ael10,?B305,?Bel03,?Bel06/?Bx02,?Bw07,?Bw12,?Bw17,?Bw37,?O05,?O26,?O61,?B(A)04,?B(A)07,?cisAB01 and ?cisAB01var).In addition,six rare allele mutation sites were identified(c.101A>G;c.103_106delG;c.146_147insGC;c.259G>T;c.322C>T;c.932T>C).Conclusion The identification of ambiguous ABO blood group requires the combination of serological testing and molecular biological examination to correctly identify the blood type and ensure the safety of clinical blood transfusion.
2.Prevalence of out-of-hospital prevalence of carbapenem-resistance genes and influences of antibiotics with different antibacte-rial mechanisms on the genes spread
Lanlan FENG ; Jinhui TAN ; Ping ZHAO ; Zhihong XU ; Yihua ZHU ; Xiang CHEN
Chinese Journal of Clinical Laboratory Science 2024;42(8):567-573
Objective To analyze the distribution of carbapenem-resistant strains and the transmission mode of resistance genes in the bacteria strains from urinary system of the individuals outside hospitals,and further explore the impacts of the antibiotics with different antibacterial mechanisms on the conjugation transfer of resistant plasmids.Methods The imipenem-resistant strains from 991 non-re-petitive urine samples of healthy individuals were screened for determining the carbapenem-resistance genotypes.Using the multilocus sequence typing(MLST)method,the main carbapenem-resistant strains were classified to determine the phylogenetic relationship be-tween different sequence types(STs)and reveal the transmission mode of resistance genes in the individuals outside hospitals.The conjugation experiment was used to analyze the effects of different sub-inhibitory concentrations of ampicillin,ciprofloxacin,and genta-micin on the conjugation efficiency of resistant plasmids in the bacteria of donor and/or recipient at different times.Results A total of 18 non-repetitive carbapenem-resistant strains(1.82%,18/991)were detected in healthy individuals outside hospitals,among which Escherichia coli(E.coli)(44.44%,8/18)was the main strain with main genotype being blaKPC-2(7/8).MLST showed that the 7 strains of E.coli with blaKPC-2 genotype belonged to ST-10,ST-101,ST-131,ST-405,ST-410,ST-1193,and ST-2562,respectively.Homologous clustering analysis showed that the type of the 7 E.coli strains exhibited high diversity.The range of joint efficiency was(1.59±0.20)× 10-4 to(4.01±1.31)× 10-4.The highest conjugation efficiency was observed in the donor bacteria treated with 1/8 of MIC of antibiotics for 9 hours,and the conjugation efficiency most significantly increased compared to the group without antibiotic treat-ment.Three antibiotics with different mechanisms of action could all improve conjugation efficiency,among which ampicillin exhibited the most significant improvement.Conclusion In this study,the genotype of carbapenem-resistant Escherichia coli(CREC)was mainly blaKPC-2 with a main mode of horizontal transmission.Antibiotics with sub-inhibitory concentrations could promote the conjugation transfer of the resistant plasmid.There may be significant differences in the effects of antibiotics with different mechanisms of action and different antibiotic stress conditions on the conjugation efficiency.
3.Rapid progressive dementia and involuntary movement caused by vitamin B12 deficiency:a case report
Lanlan LIU ; Xin JIANG ; Xuewei XING ; Yanqing TIE ; Guosong QI ; Jingjing WANG ; Xiaoxiao FENG ; Peiyuan LYU
Chinese Journal of Nervous and Mental Diseases 2024;50(3):165-167
A 41-year-old male was presented with rapidly progression memory impairment for 2 months and episodic limb shaking for 2 weeks as the main manifestations.Physical examination showed verbal disadvantage with decreased memory,attention,comprehension,and orientation.Serum vitamin B12 levels decreased,serum anti gastric parietal cell antibodies and anti-intrinsic factor antibodies were positive.Blood analysis showed macrocytic anemia,neuropsychological scale showed functional impairment in multiple cognitive domains,electrophysiological examination showed peripheral nerve damage,cerebrospinal fluid and imaging examination showed no abnormalities.The patient was diagnosed as having vitamin B12 deficiency dementia,vitamin B12 deficiency related involuntary movement and pernicious anemia.Supplementing with B vitamins and folic acid significantly improved cognitive impairment and eliminated symptoms of limb shaking.The purpose of this case report is to enhance the understanding of clinical doctors about dementia and involunting movement caused by vitamin B12 deficiency,in order to diagnose and treat it early.
4.Correlations between depression, sleep quality and dental caries among college students in Chongqing
Chinese Journal of School Health 2023;44(7):1080-1083
Objective:
To understand the correlation between depression, sleep quality and dental caries among college students in Chongqing, so as so provide reference basis for targeted health education.
Methods:
From January to June 2022, a multi stage sampling method was used to select 627 students from six colleges in Chongqing. According to the Fourth National Epidemiological Survey of Oral Health and World Health Organization standards, dental caries were examined and diagnosed. The Self Rating Depression Scale (SDS), Pittsburgh Sleep Quality Index (PSQI) Questionnaire and general situation questionnaire were adopted to investigate depressive symptoms and sleep quality of college students. Correlation analysis was performed to study the correlation between depression, sleep quality and dental caries.
Results:
The prevalence of dental caries among college students was 42.1%, and there was a statistically significant difference in dental caries prevalence among college students depending on their gender, major, body shape, parental education level, and family sources ( χ 2=14.49, 16.81, 7.82, 15.14, 7.34, P <0.05). The prevalence of depressive symptoms was 13.9%, and there was a statistically significant difference in SDS scores and depression prevalence among students of different body types ( χ 2=10.99, P <0.05). The PSQI score was (4.36±0.94) points, and the rate of poor sleep quality was 26.16%. Furthermore, there was a statistically significant difference in the PSQI score and rate of poor sleep quality among college students of different genders and body types ( χ 2=25.41, 17.59, P <0.05). There was a certain correlation between the occurrence of dental caries and poor sleep quality and depressive symptoms among college students, with contingency coefficients of 0.15 and 0.13, respectively ( P <0.05).
Conclusion
The prevalence of dental caries is high among college students in Chongqing. The occurrence of dental caries may be related to depression and sleep quality.
5.A metabolic intervention strategy to break evolutionary adaptability of tumor for reinforced immunotherapy.
Qianhua FENG ; Yutong HAO ; Shuaiqi YANG ; Xiaomin YUAN ; Jing CHEN ; Yuying MEI ; Lanlan LIU ; Junbiao CHANG ; Zhenzhong ZHANG ; Lei WANG
Acta Pharmaceutica Sinica B 2023;13(2):775-786
The typical hallmark of tumor evolution is metabolic dysregulation. In addition to secreting immunoregulatory metabolites, tumor cells and various immune cells display different metabolic pathways and plasticity. Harnessing the metabolic differences to reduce the tumor and immunosuppressive cells while enhancing the activity of positive immunoregulatory cells is a promising strategy. We develop a nanoplatform (CLCeMOF) based on cerium metal-organic framework (CeMOF) by lactate oxidase (LOX) modification and glutaminase inhibitor (CB839) loading. The cascade catalytic reactions induced by CLCeMOF generate reactive oxygen species "storm" to elicit immune responses. Meanwhile, LOX-mediated metabolite lactate exhaustion relieves the immunosuppressive tumor microenvironment, preparing the ground for intracellular regulation. Most noticeably, the immunometabolic checkpoint blockade therapy, as a result of glutamine antagonism, is exploited for overall cell mobilization. It is found that CLCeMOF inhibited glutamine metabolism-dependent cells (tumor cells, immunosuppressive cells, etc.), increased infiltration of dendritic cells, and especially reprogrammed CD8+ T lymphocytes with considerable metabolic flexibility toward a highly activated, long-lived, and memory-like phenotype. Such an idea intervenes both metabolite (lactate) and cellular metabolic pathway, which essentially alters overall cell fates toward the desired situation. Collectively, the metabolic intervention strategy is bound to break the evolutionary adaptability of tumors for reinforced immunotherapy.
7.Mechanism of Shenqi Yiliu Prescription Combined with Cisplatin on H22 Liver Cancer-bearing Mice Based on NLRP3/Caspase-1/GSDMD Pyroptosis Pathway
Mengying YANG ; Yongqiang DUAN ; Yuxin JIA ; Min BAI ; Zhongbo ZHU ; Yarong LI ; Lan MA ; Mingyu ZHANG ; Xin FENG ; Lanlan HE ; Yuping YANG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(21):114-122
ObjectiveTo explore the anti-tumor effect and mechanism of Shenqi Yiliu prescription in the intervention of pyroptosis. MethodTen male BALB/c mice were randomly selected and assigned to the blank group. The remaining 40 mice underwent the induction of the liver cancer xenograft model. After 5 days of modeling, 40 surviving mice were randomly divided into model group, cisplatin group [2.5×10-3 g·kg-1·(3 d)-1], Shenqi Yiliu prescription group (27 g·kg-1·d-1), and a combination group (Shenqi Yiliu prescription group + cisplatin). The mice in the blank group and the model group were treated with an equal volume of normal saline for 10 days. The general conditions of mice in each group were observed. After the intervention, the tumor weight of the mice was weighed and the tumor inhibition rate was calculated. Hematoxylin-eosin (HE) staining was used to observe the pathological changes in tumor tissues. The levels of mouse liver function indicators, including alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected. The TdT-mediated dUTP-biotin nick end labeling (TUNEL) assay was used to detect DNA damage in mouse tumor tissue cells. Immunohistochemistry (IHC), immunofluorescence (IF), and Western blot were used to detect the protein expression levels of NOD-like receptor protein 3 (NLRP3), cysteinyl aspartate-specific protease-1 (Caspase-1), and gasdermin D (GSDMD) in tumor tissues. The levels of interleukin-1β (IL-1β) and interleukin-18 (IL-18) in tumor tissues were detected by enzyme-linked immunosorbent assay (ELISA). ResultCompared with the mice in the blank group, those in the model group were in a poor mental state, sleepy, and lazy, and their fur color was dull, with increased levels of serum ALT and AST in liver function tests (P<0.01). Compared with the model group, the groups with drug intervention showed improved mental state, inhibited tumor growth to varying degrees, and decreased tumor weight, and the tumor inhibition rate in the combination group was the highest (P<0.01). HE staining showed that the pathological and morphological lesions of the tumor tissues in the model group were significant, while those in all groups with drug intervention were improved to a certain extent. The karyolysis and nuclear rupture in the Shenqi Yiliu prescription group and the combination group were more significant. In the liver function test, the serum ALT and AST levels of mice in the Shenqi Yiliu prescription group and the combination group decreased (P<0.01), and the inflammatory factors IL-1β and IL-18 in each group with drug intervention decreased (P<0.05, P<0.01). Among them, the declining trend of IL-1β and IL-18 in the Shenqi Yiliu prescription group was the most significant (P<0.01). TUNEL staining showed that the positive TUNEL staining in each group with drug intervention decreased after intervention (P<0.05, P<0.01), especially the cisplatin group and Shenqi Yiliu prescription group (P<0.01). Western blot, IHC, and IF found that the protein expression levels of NLRP3, Caspase-1, and GSDMD in each group with drug intervention decreased (P<0.05, P<0.01). Compared with the mice in the cisplatin group, those in the Shenqi Yiliu prescription group and the combination group had better mental state and regular tumor morphology, and the tumor weight of the mice in the combination group decreased (P<0.05). The levels of ALT and AST in the Shenqi Yiliu prescription group decreased (P<0.05), and the levels of IL-1β and IL-18 in the Shenqi Yiliu prescription group and the combination group decreased (P<0.05, P<0.01), especially in the combination group (P<0.01). The results of IHC showed that the expression of GSDMD protein in the tumor tissues of mice in the combination group was reduced (P<0.01). IF detection showed that the expression of NLRP3 in the tumor tissues of the Shenqi Yiliu prescription group was reduced (P<0.01). The results of Western blot showed that the expression level of NLRP3 protein in the Shenqi Yiliu prescription group and the combination group decreased (P<0.01), and the expression level of Caspase-1 protein in the combination group decreased (P<0.01). The decrease in GSDMD protein expression was not significant, and the difference was not statistically significant. ConclusionShenqi Yiliu prescription combined with cisplatin has an obvious anti-tumor effect, which may be achieved by down-regulating the NLRP3/Caspase-1/GSDMD inflammatory pyroptosis pathway to inhibit cell pyroptosis, and relieve the inflammatory response in mice with liver cancer.
8.Analysis of vitamin D nutritional status among children aged 0 to 12 years in Chongqing
WANG Xu, LIU Xia, ZHU Lanlan, DENG Yu, XIAO Xiang, LIU Yong, LIU Feng
Chinese Journal of School Health 2022;43(11):1638-1641
Objective:
To investigate the nutritional status and associated factors of vitamin D among children aged 0 to 12 years old in Chongqing.
Methods:
From January 2019 to December 2021, 1 877 children aged 0-6 years who received regular child health care were randomly selected from Chongqing maternal and child health hospital, and 707 school age children were also selected from three primary school in Chongqing for investigation. The serum sample were collected for detecting serum 25 (OH) D level by chemiluminescence method, and the possible influencing factors of vitamin D deficiency (VDD) were analyzed.
Results:
The average serum 25 (OH) D concentration of children was (39.9±9.1) ng/mL with the positive rate of VDD was 14.59%. There were significant differences of VDD positive rate in different age, visiting time, visiting season, body shape, outdoor activity time and vitamin D supplementation (0-3 years old vs 4-6 years old vs 7-12 years old: 14.73% vs 22.19% vs 10.47%; visiting season: spring vs summer vs autumn vs winter: 12.97% vs 10.65% vs 14.86% vs 21.33%; pre COVID 19 epidemic vs post COVID-19 epidemic: 11.18% vs 17.08%; underweight or normal vs overweight or obesity: 12.34% vs 26.13%; adequate outdoor activity time vs inadequate:11.84% vs 16.27%; regular vitamin D supplementation vs irregular vitamin D supplementation: 11.71% vs 15.62% )( χ 2=26.17, 17.59, 30.98, 53.74, 9.60, 6.17, P <0.05). Underweight or normal body weight, sufficient outdoor activities and regular vitamin D supplementation were associated with less vitamin D deficiency with OR(OR95%CI ) at 0.68(0.55-0.84), 0.57 (0.25-0.77) and 0.62(0.44-0.85)( P <0.05).
Conclusion
The prevalence of VDD among preschool and school age children is high in Chongqing. Integrated prevention and control strategies incluing overweight or obesity control, increasing outdoor activities and oral vitamin D supplements should be taken.
9.Primary diffuse large B-cell lymphoma in cavernous sinus: report of 1 case and review of literature
Conggang HUANG ; Ming LUO ; Liang ZOU ; Lanlan WANG ; Yuhan BAI ; Yan FENG ; Jinglei WU ; Xingwan WANG ; Zhihua LUO ; Faliang DUAN
Journal of Leukemia & Lymphoma 2022;31(9):551-555
Objective:To investigate the clinicopathological features, treatment programs and prognosis of patients with primary diffuse large B-cell lymphoma (DLBCL) in cavernous sinus.Methods:The clinical data of a patient with primary DLBCL in cavernous sinus who were admitted to Wuhan No.1 Hospital in December 2020 were retrospectively analyzed, and the relevant literature was reviewed.Results:The patient was a 63-year-old female who underwent resection of the cavernous sinus lesion, and the pathological diagnosis was DLBCL. The patient received 6 courses of R-CHOP regimen chemotherapy, lumbar puncture + intrathecal injection of chemotherapy drugs, and twice additional rituximab immunochemotherapy, and no tumor cells were found in the results of liquid-based thin layer cytology for cerebrospinal fluid exfoliated cells; twice magnetic resonance imaging (MRI) re-examination after the operation showed no recurrence and adjacent metastasis of the tumor. The patient's symptoms were significantly improved without residual neurological sequelae.Conclusions:Primary DLBCL in cavernous sinus is rare in clinical practice, early diagnosis is crucial for the prognosis of patients, and different protein expression may indicate the prognosis. Biopsy, complete resection of the tumor under the premise of preserving important anatomical structures and functions, and standardized chemotherapy combined with intrathecal injection local chemotherapy can effectively prolong the survival time of patients and improve the quality of life.
10.Comparative analysis of clinical features between giant cell arteritis and polymyalgia rheumatica
Dandan FENG ; Xiaowan WANG ; Lanlan JIA ; Ruijun ZHANG ; Liang XU
Chinese Journal of Rheumatology 2021;25(2):114-117
Objective:To identify the differences in clinical manifestations between Chinese giant cell arteritis (GCA) patients and polymyalgia rheumatica (PMR) patients.Methods:Twelve GCA patients were included during September 2010 to September 2019 in our hospital. Clinical and laboratory data were collected. Twenty-four age and sex-matched pure PMR patients were selected as control. Statistical analysis was performed using Statistical product and service solutions (SPSS) software. The categorical variables were tested by chi square test, and the continuous variables were expressed by mean and standard deviation ( ± s). The comparison between groups was conducted by t-test. P<0.05 was considered statistically significant. Results:In these 12 GCA patients, the onset age was 55-70 (67±7) years old, and male to female ratio was 1∶11. The most common initial symptom of GCA was the same as PMR (7/12, 58%) . Compared with PMR patients, the specific clinical manifestations of GCA patients were scalp pain ( P=0.031), mandibular claudication ( P=0.031) and migraine ( P=0.000). The creatine kinase of GCA (60±27) U/L patients was higher than that in PMR (41±15) patients ( t=1.098, P=0.029). There was no significant difference in other laboratory tests including erythrocyte sedimentation rate, C reactive protein level. Seven of 12 GCA patients were first diagnosed with PMR, then were diagnosed with GCA during follow-up. No obvious differences could be found in clinical manifestations between these 7 patients and 24 pure PMR patients. Through imaging examinations, we found that 9 of the 12 GCA patients had arterial stenosis, 5 had thickened vascular walls, 5 had atherosclerosis, and 2 had rough endometrium. Conclusion:GCA patients and PMR may have similar clinical presentations. The presence of scalp pain, mandibular claudication and migraine during the course of the disease implies that GCA is more likely. Vascular ultrasound, arterial CTA, and positron emission tomograph (PET)/CT play an increasingly important role in the diagnosis of GCA.


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