1.Viable Bacteria Assay of Helicobacter pylori by RT-qPCR Measurement of cgt Gene Expression Levels:Establishment and Application of a New Method
Zhihui TANG ; Lifa FU ; Yanrong ZHANG ; Boyan ZHOU ; Tianqin FENG ; Wenjuan YANG ; Ge LIANG ; Qianya YAN ; Canlin ZHENG ; Mingjiang BIE ; Baoning WANG
Journal of Sichuan University (Medical Sciences) 2024;55(5):1316-1321
Objective To establish a viable bacteria assay for Helicobacter pylori(H.pylori)by assessing the cgt gene expression,and to develop accordingly a rapid and novel testing method for clinical precision treatment.Methods Viable bacteria count was determined in bacterial cultures.The transcriptional expression level of cgt(hp0421),the conserved gene that encodes cholesterol-α-glucosyltransferase(CGT)in H.pylori,was measured by RT-PCR.The correlation between the number of colonies and cgt gene transcription expression was analyzed and the regression model was constructed.The linear range,sensitivity,and specificity of the new method were examined accordingly.The bactericidal action of clarithromycin was assessed using this method to verify the performance of the method in determining clinical bacterial drug resistance.Results The Ct values of cgt for H.pylori colony counts of 102,104,106,and 108 CFU/mL were 29.67±0.14,23.37±0.36,17.65±0.37,and 11.38±0.39,respectively.In the range of 101-108 CFU/mL,the regression equation for cgt gene expression and viable bacterial counts determined by RT-qPCR was y=-0.350 1x+12.49,with the correlation coefficient being R2=0.9992 and the sensitivity being 101 CFU/mL,showing no cross-reaction with 13 other bacteria.The lg values of live H.pylori bacteria treated with clarithromycin at 0,5,10,20,and 40 μg/mL for 12 h were 2.57±0.02,2.45±0.01,2.19±0.02,1.91±0.07,and 1.33±0.05,respectively.The corresponding cgt gene Ct values were 27.76±0.09,28.37±0.24,29.51±0.14,30.11±0.12,and 31.66±0.11.By applying the cgt gene expression in the equation,the estimated counts of viable bacteria were found to be 2.73±0.03,2.52±0.08,2.11±0.05,1.89±0.02,and 1.33±0.04,showing no significant difference in statistical analysis(P>0.05).Conclusion The method for assessing viable bacteria account by evaluating cgt gene expression in H.pylori was successfully established,significantly reducing the time required to determine viable bacteria count and providing a new method for clinical viable bacteria testing.
2.Assessment of renal function and renal morphology in preterms and term newborns
Qianya XU ; Qian ZHANG ; Xinru CHENG ; Xiao HE ; Li WANG ; Shanshan ZHANG ; Zanyang SHI ; Nan LIU ; Xiaomin YAN ; Jianguo WEN
Chinese Journal of Applied Clinical Pediatrics 2019;34(7):525-528
Objective To explore the relationship between renal volume and function in the preterm and term newborns at different gestational ages.Methods This study was conducted at the First Affiliated Hospital of Zhengzhou University from October 2016 to March 2018.A total of 626 newborns with different gestational ages were included and the renal volume was determined by ultrasonography.Blood samples were taken for laboratory tests to detect renal function including urea,creatinine,uric acid and estimated glomerular filtration rate.Results A total of 352 preterm and 274 full term newborns were enrolled at birth.The mean gestational age of the neonates was (36.0 ± 3.5) weeks and the mean birth weight was (2.59 ±0.77) kg.The mean renal volume of the preterm infants was (19.57 ±4.30) cm3 and estimated glomerular filtration was (21.68 ± 5.99) mL/(min · 1.73 m2);the mean renal volume of the term infants was(23.03 ± 3.80) cm3 and estimated glomerular filtration was(46.60 ± 10.21) mL/(min · 1.73 m2).The renal volume and estimated glomerular filtration of term infants was significantly greater compared to the preterm infants (t =12.96,33.10,all P < 0.001).The renal volume was highly linear positively correlated with gestational age,birth weight and birth length (r =0.546,0.605,0.592,all P < 0.001).The renal volume was highly linear positively correlated with estimated glomerular filtration(r =0.396,P < 0.001).The renal volume was negatively correlated with urea,creatinine and uric acid(r =-0.210,-0.280,-0.176,all P < 0.001).Conclusions The renal volume increases with gestational age and birth weight in neonates.Estimated glomerular filtration increases with renal volume in neonates.The preterm infants have immaturity kidney size and poor development so that they need special medical care.
3.Analysis of testis development and epididymal morphology in preterm and term newborns
Qianya XU ; Jianguo WEN ; Xinru CHENG ; Shanshan ZHANG ; Zanyang SHI ; Nan LIU ; Xiao HE ; Yan LI ; Qian ZHANG
Chinese Journal of Applied Clinical Pediatrics 2019;34(8):609-613
Objective To explore the testis development and epididymis in the preterm and term newborns so as to provide the scientific evidence for early clinical diagnosing early.Methods From October 2016 to March 2018,456 hospitalized neonates at Department of Neonatology,the First Affiliated Hospital of Zhengzhou University were recruited within 7 days at birth in this study.In these patients,224 cases were preterm newborns and 232 cases were term newborns.These gestational ages of newborns at birth were (36.18 ± 3.13) weeks (27-41 +6 weeks) and weighted (2.66 ± 0.67) kg(0.90-3.82 kg).The size of the testis and epididymis were measured by ultrasonography.Results The mean testicular volume of the preterm was (0.24 ± 0.07) mL.The mean length of epididymal head,thickness of epididymal head and body and tail of the preterm newborns were (4.17 ±0.59) mm,(2.58 ±0.39) mm,(1.78 ± 0.26) mm,(1.91 ± 0.24) mm,respectively.The mean testicular volume of the term newborns was (0.38 ± 0.13) mL,the mean length of epididymal head,thickness of epididymal head and body and tail of the term newborns were (4.49 ± 0.45) mm,(2.78 ± 0.34) mm,(1.95 ± 0.20) mm,(1.99 ± 0.16) mm,respectively.The mean testicular volume,length of epididymal head,thickness of epididymal head and body and tail of the preterm newborns were significantly lower compared with the term newborns (t =12.810,8.261,6.819,8.058,3.591,all P <0.001).The mean testicular volume of the newborns were highly linear positively correlated with gestational age,birth weight and birth length (r =0.538,0.591,0.533,all P < 0.001).In the preterm newborns at postmenstrual age (PMA) of 37 weeks,the mean testicular volume,length of epididymal head,thickness of epididymal head and body and tail of the preterm newborns had no significant difference between the 2 groups (t =1.561,0.863,0.282,1.732,1.147,all P > 0.05).Conclusions The testicular volume,the length of epididymal head,thickness of epididymal head and body increase with gestational age,birth weight and birth length in early neonates.The growth of reproductive system in the preterm newborns at PMA 37 weeks catch-up with term newborns.If this catch-up growth was incomplete at PMA 37 weeks,special attention should be given to monitor underlying diseases.

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