1.Development of multiple primer, HIV mini-pool NAT and its application in detecting acute infection of MSM
Yasong WU ; Zhiying LIU ; Yanmei JIAO ; Feili WEI ; Meng XU ; Tong ZHANG ; Xiaojie HUANG ; Fujie ZHANG ; Dexi CHEN ; Xiaoning XU ; Hao WU
Chinese Journal of Laboratory Medicine 2010;33(9):862-866
Objective To establish a mini-pool nucleic acid testing (NAT) assay using multiplex RT-nested PCR for the detection of HIV RNA, and apply it in screening for acute HIV infection among MSM. Methods Frozen EDTA plasma samples collected between Oct. 2008 and Mar. 2009 from 3 HIV infectors during window-period, a total of 30 HIV chronically infected individuals and 97 healthy subjects were used to develop the NAT assay. Plasma samples from 10 cases were pooled into one tube and centrifuged at high speed for the collection of viruses. HIV RNA was extracted. Two pairs of primers were designed according to two conserved regions of HIV RNA ( HXB2 nt 5783-nt 6228 and nt 1235-nt 2012).Multiplex RT-PCR and nested PCR were performed. Individual NAT-reactive samples were confirmed by commercially available NAT assays. The sensitivity and performance efficacy were also evaluated. The assay was then applied to 1 005 plasma specimens from MSM with negative or uncertain HIV antibody test results.These were collected in the same period as the other samples. Results ( 1 ) Two fragments of HIV were amplified successfully with the low detection limit of 162 copies/ml plasma; (2) Results of the mini-pool HIV NAT validation with samples from 3 HIV infectors during window-period were consistent with the expected values; (3) All 30 plasma samples from MSM with positive HIV antibody, which were tested by multiplex RT nested PCR, were found to be HIV RNA positive; (4) One out of 1 005 plasma samples was found to be HIV RNA positive, for this case acute infection was followed-up and sero-conversion was found. Conclusion Mini-pool NAT has good sensitivity, and may be applied to screening HIV RNA among MSM during window-period.
2. Diagnosis of a fetus with X-linked hydrocephalus due to mutation of L1CAM gene
Qichang WU ; Li SUN ; Yasong XU ; Xiaomei YANG ; Shiyu SUN ; Wenbo WANG
Chinese Journal of Medical Genetics 2019;36(9):897-900
Objective:
To explore the genetic basis for a case of recurrent fetal congenital hydrocephalus.
Methods:
Next-generation sequencing was carried out for the fetus, the gravida and two of her sisters.
Results:
The fetus was found to harbor a c. 1765T>C (p.Tyr589His) mutation in exon 14 of the L1CAM gene, which was derived from the gravida.
Conclusion
Male fetuses with recurrent hydrocephalus should be subjected to testing of the L1CAM gene to facilitate genetic counseling and prenatal diagnosis.
3.Application value of procalcitonin clearance rate on clinical outcome in patients with severe pneumonia
Guijun ZHENG ; Jiegen ZHANG ; Yasong YUAN ; Duoduo XU ; Shengnan DONG ; Huifang WANG
Chinese Critical Care Medicine 2019;31(5):566-570
Objective To investigate the changes in serum procalcitonin (PCT) in patients with severe pneumonia, and to analyze its value on evaluating the clinical outcome of patients with severe pneumonia. Methods A total of 58 patients with severe pneumonia aged over 18 years, and admitted to intensive care unit (ICU) of Zhuozhou City Hospital of Hebei Province from January 2017 to July 2018 were enrolled. The patients were divided into recovery group (the symptoms and signs of pneumonia disappeared or improved, and the X-ray chest films improved or did not make significant progress) and deterioration group (the symptoms and signs of pneumonia persisted or progressed, while X-ray chest radiography progressed, as well as serious complications such as involvement of other organ functions due to deterioration of pulmonary infection or septic shock) according to the therapeutic outcome. The serum PCT levels at 1, 3, 5, 7, 9 days after severe pneumonia diagnosed were recorded, and procalcitonin clearance rate (PCTc) was calculated. The acute physiology and chronic health evaluation Ⅱ (APACHEⅡ) score was estimated within 24 hours when severe pneumonia was diagnosed. Receiver operating characteristic (ROC) curve was drawn, and the area under ROC curve (AUC) was calculated to analyze the value of PCTc on evaluating the clinical outcome of patients with severe pneumonia. Results Among 58 patients, 33 (56.9%) had better outcome after active treatment (recovery group), and 25 (44.1%) had worse condition (deterioration group). There was no significant difference in PCT level at 1 day or 3 days between the recovery group and the deterioration group [μg/L: 5.05 (3.89, 7.61) vs. 5.29 (4.15, 7.46) at 1 day, 4.59 (4.02, 6.90) vs. 5.70 (4.59, 7.28) at 3 days, both P > 0.05]. With the prolongation of treatment time, serum PCT level was gradually decreased in the recovery group, while remained at higher level in the deterioration group, which was significantly lowered at 5, 7, 9 days in the recovery group as compared with that in the deterioration group [μg/L:2.92 (2.09, 3.42) vs. 6.09 (3.24, 7.96) at 5 days, 1.94 (1.50, 2.07) vs. 7.65 (5.60, 10.52) at 7 days, 1.37 (0.91, 1.74) vs. 8.96 (6.09, 10.87) at 9 days, all P < 0.01]. PCTc at 3, 5, 7, 9 days in the recovery group were significantly higher than those in the deterioration group [15.10 (-17.80, 32.10)% vs. -1.53 (-20.80, 11.48)% at 3 days, 47.50 (30.25, 60.34)% vs. 6.25 (-14.58, 29.05)% at 5 days, 76.44 (53.18, 77.92)% vs. -11.20 (-66.75, -1.38)% at 7 days, 80.01 (59.86, 88.27)% vs. -38.15 (-99.38, -2.81)% at 9 days, all P < 0.05]. ROC curve analysis showed that PCTc at 3, 5, 7 and 9 days were valuable for evaluating the clinical outcome of patients with severe pneumonia, and 9-day PCTc had the greatest value, the AUC was 0.978 [95% confidence interval (95%CI) = 0.945-1.000, P = 0.000], which was higher than APACHEⅡ(AUC = 0.442, 95%CI = 0.280-0.610, P = 0.392); when the best cut-off value of 9-day PCTc was 93.00%, its sensitivity was 99.0%, and specificity was 87.3%. Conclusions The PCT level of patients with severe pneumonia remained at a high level, which was related with the deterioration of the disease. PCTc, as an index to evaluate the clinical outcome of patients with severe pneumonia, has good application value.
4.Diagnosis of a fetus with X-linked hydrocephalus due to mutation of L1CAM gene.
Qichang WU ; Li SUN ; Yasong XU ; Xiaomei YANG ; Shiyu SUN ; Wenbo WANG
Chinese Journal of Medical Genetics 2019;36(9):897-900
OBJECTIVE:
To explore the genetic basis for a case of recurrent fetal congenital hydrocephalus.
METHODS:
Next-generation sequencing was carried out for the fetus, the gravida and two of her sisters.
RESULTS:
The fetus was found to harbor a c.1765T>C (p.Tyr589His) mutation in exon 14 of the L1CAM gene, which was derived from the gravida.
CONCLUSION
Male fetuses with recurrent hydrocephalus should be subjected to testing of the L1CAM gene to facilitate genetic counseling and prenatal diagnosis.
DNA Mutational Analysis
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Female
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Fetus
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Genetic Diseases, X-Linked
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diagnosis
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genetics
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Humans
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Hydrocephalus
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diagnosis
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genetics
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Male
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Mutation
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Neural Cell Adhesion Molecule L1
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genetics
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Pedigree
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Pregnancy
5.Social skill training for children with autistic spectrum disorder on the uncertainty of the disease from mothers
ZU Yanfei, DU Yasong, ZHOU Yingqun, FAN Ning, ZHU Shuyi, CAO Yang, JIANG Wenqing, XU Guangxing
Chinese Journal of School Health 2022;43(8):1249-1253
Objective:
To improve the social skills of children with ASD by using Program for the Education and Enrichment of Relational Skills(PEERS ), and to reduce the uncertainty towards ASD and negative emotions for mothers of ASD children.
Methods:
From September to October 2017, 30 dyads of autistic mother and child were recruited and divided into intervention group and control group (15 mother child dyads each). Based on the content of PEERS social skill, cognitive behavior therapy was delivered in group format, through demonstration, role play and group exercise. At the same time, mother child dyads were trained using parallel social technology. Mothers and children with ASD were investigated using Parents Perception of Uncertainty Scale (PPUS), Patient Health Questionnaire 9 (PHQ-9), Chinese Version of the Beck Depression Inventory II(BDI-Ⅱ-C), Beck Anxiety Inventory (BAI), State Trait Anxiety Inventory(STAI-Form Y), and Autism Behavior Checklist (ABC), Cildhood Autism Rating Scale (CARS), and Social Communication Questionnaire (SCQ).
Results:
Changes in ASD symptom score in children and emotional score of mothers in the intervention group were less than 0. The total score of mother disease uncertainty(74.93±13.58, 90.40± 9.21 ), ambiguity(31.13±7.07, 38.93±4.73), lack of clarity information(11.93±2.09, 13.80±2.54), unpredictability(9.60±1.99, 12.07±2.89), significantly changed after intervention( t =-3.65, -3.55, -2.20, -2.72, P <0.05).
Conclusion
Social PEERS group intervention can enhance the social skills of children with ASD, reduce uncertainty of illness among mother of ASD children. Timely disease related information, guidance for mothers to actively participate in child care and training, might help to reduce cognitive bias, depressive and anxiety symptoms among mothers.
6.Effect of autophagy response on neurological functions and its mechanism in rats after traumatic brain injury
Xiangrong CHEN ; Zhihui TANG ; Yasong LI ; Qinliang LUO ; Weipeng HU ; Chaoyang XU
Chinese Journal of Neuromedicine 2016;15(12):1200-1205
Objective To investigate the effect of autophagy response on neurological functions and the role of mitogen-activated protein kinases (MAPKs) signaling pathway in rats after traumatic brain injury (TBI).Methods Fifty-four healthy male SD rats were randomly divided into sham-operated group,TBI group and TBI+autophagy inhibitor 3-methyladenine (3-MA) group (n=18).TBI animal models of the later two groups were established using Feeney's method.Rats in the sham-operated group were only performed bone window opening without knock;rats in the TBI+3-MA group were given intraperitoneal injection of 3-MA(5 mg/kg) 30 min after modeling and rats in the other two groups were given the same volume of normal saline.Three and 7 d after modeling,the protein levels of S100B and neuron specific enolase (NSE) in serum were tested with enzyme linked immunosorbent assay (ELISA);modified neurologic severity scale (mNSS) was used to detect the movement,sense and reflex functions;brain water content was measured with wet-dry weight method.The autophagy related factors (LC3-Ⅱ and Beclin-1) and MAPKs signaling pathway related factors (c-Jun N-terminal kinase [JNK],phosphorylated [p]-JNK,extracellular signal-regulated kinase [ERK]1/2,p-ERK1/2,p38MAPK and p-p38MAPK) protein expressions in TBI cerebral cortex were determined by Western blotting.Results As compared with those in the sham-operated group,the brain edema level,mNSS scores,and S100B and NSE protein levels in the TBI group and TBI+3-MA group were significantly increased (P<0.05);TBI+3-MA group had significantly lower brain edema level,mNSS scores,and S100B and NSE protein levels than TBI group (P<0.05).The expression levels of autophagy and MAPKs signaling pathway related factors in the TBI group and TBI+3-MA group were significantly higher as compared with those in the sham-operated group (P<0.05).As compared with the TBI group,TBI+3-MA group had significantly decreased levels of LC3-Ⅱ,Beclin-1 and activation of JNK and p-p38MAPK signaling pathways (P<0.05).Conclusion Suppressing autophagy response markedly improves neurological outcomes after TBI,possibly mediated by inhibiting activation of JNK and p38MAPK signaling pathways.
7.Clinical features and genetic analysis of two fetuses with ring chromosome 21 mosaicism.
Yizhen JI ; Yasong XU ; Li SUN ; Yunsheng GE ; Meijiao CAI ; Qichang WU
Chinese Journal of Medical Genetics 2023;40(8):1032-1035
OBJECTIVE:
To investigate the perinatal clinical phenotype and genetic characteristics of two fetuses with ring chromosome 21 mosaicisms.
METHODS:
Two fetuses who were diagnosed at the Xiamen Maternal and Child Health Care Hospital in November 2021 were selected as the study subjects. Clinical data of the two fetuses were collected. Conventional G-banded karyotyping and chromosomal microarray analysis (CMA) were carried out for the fetuses and their parents.
RESULTS:
Prenatal ultrasonography of fetus 1 has revealed absence of nasal bone, ventricular septal defect, persistent left superior vena cava, and mild tricuspid regurgitation. Chromosomal karyotyping was 46,X?,dic r(21;21)(p12q22;q22p12)[41]/45,X?,-21[9]. CMA has revealed a 30.00 Mb quadruplication at 21q11.2q22.3 and a 3.00 Mb deletion at 21q22.3. For fetus 2, ultrasonography has revealed pointed echo of the nasal bone. The fetus was found to have a karyotype of 46,X?,r(21)(p12q22)[83]/45,X?,-21[14]/46,X?,dic r(21;21)(p12q22;q22p12)[3]. CMA has revealed a 5.10 Mb quadruplication at 21q22.12q22.3 and a 2.30 Mb deletion at 21q22.3.
CONCLUSION
The perinatal phenotype of the two fetuses with ring chromosome 21 mosaicisms is related to the duplication of chromosomal segments near the breakpoints of the chromosomal deletions. The combined chromosomal karyotyping and CMA has enabled prenatal diagnosis and genetic counseling for these families.
Pregnancy
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Female
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Humans
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Mosaicism
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Ring Chromosomes
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Vena Cava, Superior
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Chromosome Aberrations
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Prenatal Diagnosis
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Microarray Analysis
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Fetus/diagnostic imaging*
8.Optimization of microwave extraction condition of Gynura divaricata polysaccharide by response surface analysis
Yasong ZHU ; Wei XU ; Rong SHAO ; Yuezhao ZHU ; Delin Cheng ; Chuanhua LIAO
Journal of China Pharmaceutical University 2016;47(3):359-362
Gynura divaricata polysaccharide was extracted from Gynura divaricata with the method of microwave-assisted extraction. Based on the results of single factor experiments, the extraction process was optimized by response surface analysis. The results indicated that the influencing factors during extraction were: liquid-solid ratio, microwave time, and microwave power, among which the influence of liquid-solid ratio was the most significant. The optimal conditions were as follows: liquid-solid ratio of 45∶1(mL/g), microwave power of 390 W, microwave time of 22 min. Under optimal extraction conditions, the extraction rate of Gynura divaricata polysaccharide was 15. 36%, which was pretty close to the predicted result. The optimized response surface methodology had some practical value for the process.
9.Effects of Augmented Renal Clearance on Blood Trough Concentration of Patients Receiving High-dose Regimen of Teicoplanin
Lian TANG ; Lu SHI ; Hongzhi XUE ; Zhiwei ZHUANG ; Yunlong YUAN ; Chunxia QIAN ; Jinwei ZHU ; Xiaowen XU ; Yasong ZHU ; Jisong LIU ; Yi SHEN ; Jian LU
China Pharmacy 2020;31(21):2650-2655
OBJECTIVE:To study the effects of augmented renal clearance (ARC)on blood trough concentration of patients receiving high-dose regimen of teicoplanin. METHODS :Patients who received high-dose regimen of teicoplanin in the ICU were prospectively collected from the Affiliated Suzhou Hospital of Nanjing Medical University/Suzhou Municipal Hospital during Jul. 2018-Jun. 2020. They were divided into ARC group and normal renal function group according to corrected creatinine clearance. The dosage regimen of teicoplanin in the two groups were loading dose of 600 mg,q12 h×3 doses,maintenance dose of 6-10 mg/kg,qd,and the dosage was adjusted in combination with creatinine clearance rate and blood trough concentration. The trough concentration of blood samples which were collected 30 min before the 4th and 8th-10th dosage of teicoplanin were determined by HPLC. Trough concentration ,clinical efficacy ,Gram-positive bacterial clearance rate and the occurrence of ADR were compared between 2 groups. RESULTS :A total of 56 patients were included and divided into ARC group (18 cases)and normal renal function group (38 cases). ARC group had younger age (P<0.001)and lower serum albumin level (P=0.025)than normal renal function group. The trough concentrations before administration of the 4th and 8th-10th dosage in ARC group were lower than normal renal function group (P=0.034;P=0.035). The trough concentrations in the ARC group and normal renal function group before 8th-10th dosage were all higher than 30 min before the 4th dosage (P=0.003;P<0.001). The clinical efficacy rate and the clearance rate of Gram-positive bacteria in ARC group were 77.8% and 76.2%,which were lower than those of the normal renal function group ,but there was no statistical difference (P=0.195;P=0.223). There was no liver function damage ,hemocytopenia and allergic reaction in both groups ,but in the normal renal function group ,the causal relationship between acute renal damage and teicoplanin was assessed as “very likely ”in one patient. CONCLUSIONS :ARC patients are younger ,most of them have hypoproteinemia,and the blood trough concentrations of teicoplanin in high-dose regimen are significantly lower than those of normal renal function patients. For critical ill ARC patients ,it is advisable to increase the loading dose of teicoplanin to make the trough concentration reach the target concentration range quickly.