This study analyzed the clinical and laboratory data of 3 children diagnosed with mitochondrial disease-associated primary adrenal insufficiency (PAI) at the Guangzhou Women and Children′s Medical Center, Guangzhou Medical University from October 2018 to November 2023.All patients were normal at birth but gradually developed symptoms and were diagnosed with PAI between the ages of 1 year and 1 month and 7 years and 3 months.The children presented typical clinical symptoms of PAI, including skin and mucosal hyperpigmentation (3 cases), electrolyte disturbances (3 cases), and hypoglycemia (2 cases), as well as multisystem abnormalities related to mitochondrial disease, including recurrent infections, growth retardation, cachexia, and hyperlactatemia.Genetic testing revealed significant single deletions in mitochondrial DNA in all patients: Patient 1: m.11219_15954del, Patient 2: m.8483_13459del, and Patient 3: m.8649_16084del.Treatment with Hydrocortisone acetate replacement therapy improved the electrolyte disturbances and hypoglycemia, but issues with recurrent infections, growth retardation, and cachexia persisted.This study suggests that in clinical practice, the possibility of mitochondrial disease should be highly suspected when PAI patients present with multisystem abnormalities, especially in conjunction with hyperlactatemia.

This study analyzed the clinical and laboratory data of 3 children diagnosed with mitochondrial disease-associated primary adrenal insufficiency (PAI) at the Guangzhou Women and Children′s Medical Center, Guangzhou Medical University from October 2018 to November 2023.All patients were normal at birth but gradually developed symptoms and were diagnosed with PAI between the ages of 1 year and 1 month and 7 years and 3 months.The children presented typical clinical symptoms of PAI, including skin and mucosal hyperpigmentation (3 cases), electrolyte disturbances (3 cases), and hypoglycemia (2 cases), as well as multisystem abnormalities related to mitochondrial disease, including recurrent infections, growth retardation, cachexia, and hyperlactatemia.Genetic testing revealed significant single deletions in mitochondrial DNA in all patients: Patient 1: m.11219_15954del, Patient 2: m.8483_13459del, and Patient 3: m.8649_16084del.Treatment with Hydrocortisone acetate replacement therapy improved the electrolyte disturbances and hypoglycemia, but issues with recurrent infections, growth retardation, and cachexia persisted.This study suggests that in clinical practice, the possibility of mitochondrial disease should be highly suspected when PAI patients present with multisystem abnormalities, especially in conjunction with hyperlactatemia.

Objectives:To explore the GAA varient spectrum and the genotype-phenotype correlations in patients with glycogen storage disease type Ⅱ (Pompe disease, PD), as well as to estimate the disease incidence based on carrier rate of GAA varients in Guangzhou population.Methods:A total of 57 PD cases were retrospectively enrolled at Guangzhou Women and Children′s Medical Center from January 1, 2010 to May 31, 2020. All patients presented symptoms before the age of 18 years. Each diagnosis was further confirmed by GAA enzyme activity and GAA variants. The carrier rate of GAA varients was calculated based on variants detected by whole exon sequencing among 2 395 healthy children in Guangzhou.Results:Among the 57 PD patients (including male 26, female 31),twenty-eight patients with infantile onset PD (IOPD) presented with progressive general muscle weakness and cardiomyopathy. The mean ages of symptom onset and diagnosis were (2.5±1.4) and (5.0±3.0) months, respectively. Twenty-six cases died in the first year after birth.Twenty-three patients with late onset PD (LOPD) presented with progressive muscle weakness. Seven of them had respiratory failure at diagnosis. The mean ages of symptom onset and diagnosis were (12.0±5.0) and (17.0±7.5) years, respectively. Six children with atypical IOPD showed motor delay, muscle weakness and cardiomyopathy. Their diagnosis was confirmed at 2.5-7.0 years of age. Among the 57 patients, 47 different variants were identified in the GAA gene. Three variants: c.797C>T, c.1109G>A and c.1757C>T were novel. c.1935C>A (25/114, 21.9%) and c.2238G>C (15/114, 13.2%) were the most common variants, detected in 57.1% of IOPD and 65.2% (15/23) of LOPD patients, respectively. Among the 28 IOPD patients, 26 cases (92.9%) carried at least one missense variant which indicated positive cross-reactive immunologic material (CRIM). The carrier rate of pathogenic variants in GAA gene among healthy children was 24/2 395. The estimated incidence of PD in this population is about 1/40 000. The frequencies of pseudodeficiency variants c.1726G>A and c.2065G>A homozygotes were 26.3% (15/57) and 35.1% (20/57) in PD patients, which were significantly higher than those (1.7% (40/2 395) and 3.9% (94/2 395)) in healthy children (χ2=151.2, 121.9; both P<0.01). Conclusions:PD presents as a spectrum, some as atypical IOPD. The c.1935C>A and c.2238G>C are common variants, correlated with IOPD and LOPD respectively. The c.796C>T and c.1082C>T are usually found in atypical IOPD. The majority of IOPD patients is predicted to be CRIM positive. The estimated incidence of PD is about 1/40 000.

Objective:To evaluate and improve the performance of the newborn screening program for primary carnitine deficiency (PCD) based on tandem mass spectrometry and to investigate the incidence of PCD and molecular characteristics of SLC22A5 gene in Guangzhou.Methods:A total of 200 180 neonates born in Guangzhou from 2015 to 2019 were enrolled into the newborn screening program for PCD by tandem mass spectrometry at Guangzhou Newborn Screening Center. The positive results of screening for PCD was defined as free carnitine (C0) less than 10 μmol/L with decreased acylcarnitine species in dried blood spots of three to seven days after birth. Screen-positive newborns and their mothers were recalled for another blood spot sample. The diagnosis was confirmed based on decreased levels of C0 and acylcarnitine species in recalled blood spots and genetic analysis in SLC22A5 gene sequencing. The utility of using the sum of propionylcarnitine and palmitoylcarnitine (C3+C16) as a biomarker for acylcarnitine species in newborn screening was retrospectively evaluated. The levels of C0 and (C3+C16) at first screening were compared between newborns with PCD and newborns born to mothers with PCD by independent t test. The variant spectrum and known pathogenic variants carrier rate of SLC22A5 in 2 395 healthy children in Guangzhou Women and Children′s Medical Center through whole exon sequencing were analyzed. Results:Among 200 180 neonates, 239 (0.12%) cases were screen-positive for PCD. A total of 37 patients including 15 newborns and 22 mothers had confirmed PCD. The incidence of PCD was 1/13 345 in newborns and 1/9 099 in mothers, respectively. The positive predictive value of this program was 15.5%. Taking cutoff values of C0<8.5 μmol/L or C0 8.5~9.9 μmol/L with (C3+C16)<2 μmol/L, the number of screen-positive cases would be reduced from 810 to 224 without additional false negative case, when compared with cutoff value C0<10 μmol/L only. Both levels of C0 and (C3+C16) at first screening were not significant difference between newborns with PCD and newborns born to mothers with PCD ((6.2±2.4) vs. (5.0±1.8) μmol/L, (1.4±0.4) vs. (1.2±0.5) μmol/L, t=3.826, 0.326; P=0.058, 0.572). Seven PCD mothers experienced moderate fatigue and dizziness in the morning. One of them presented with cardiomyopathy in pregnancy. Genetic analysis of the SLC22A5 gene showed that p.S467C, p.F17L, p.R254X were the three most common variants in newborns with PCD. In PCD mothers and healthy children, the p.S467C, p.F17L and R399W were the three most common whereas the severe variant p.R254X was rare. The population carrier rate for pathogenic variants was 1 in 65 and the estimated incidence of PCD was about 1/16 500. Conclusions:Newborn screening can detect PCD both in newborns and mothers. Adding a quantitative biomarker (C3+C16) <2 μmol/L into the newborn screening program can improve the PCD screen performance. The severe variant p.R253X was common in PCD newborns but rare in PCD mothers and healthy children, indicating that the current screening program maybe failed to detect all PCD newborns and under-estimated the incidence rate of PCD in Guangzhou.

Objective: To investigate the profiles of blood amino acid and acylcarnitine in early neonates with neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD) and the sensitivity of newborn screening, and to explore potential biochemical metabolic markers for newborn screening program. Methods: Amino acid and acylcarnitine profiles in dried blood spots of newborn screening program were analyzed by tandem mass spectrometry (MS/MS). A total of 158 651 neonates born in Guangzhou from January 1, 2015 to June 30, 2019 were enrolled in this newborn screening program, and additionally 55 patients with NICCD confirmed by SLC25A13 gene analysis in Guangzhou Women and Children Medical Center were included in this study. NICCD screen-positive was defined as the cutoff value of citrulline (Cit) ≥ 30 μmol/L. The values of blood sampling time of the true positive group and those of the false negative group were compared by t-test. The levels of amino acid and acylcarnitine among different groups, including true positive group (Cit≥30 μmol/L), false negative group (Cit 21-<30 μmol/L and Cit<21 μmol/L) and the normal control group, were analyzed by F test, respectively. Results: Among 158 651 neonates, 39 neonates were positive for NICCD screening. Three of them were confirmed NICCD and 4 cases were found to be false negatives. The positive predictive value was 7.7% and the sensitivity was about 43.0%. Among 55 patients with NICCD, 18 cases (18/55, 32.7%) were true positives and 37 cases (37/55, 67.3%) were false negatives based on the cutoff value of citrulline in the dried blood spots for newborn screening. The blood sampling time was significantly different between true positive group and false negative group ((4.28±1.6) vs. (2.98±0.74) d, t=4.06, P<0.01). The increased levels of tyrosine((176.0±98.4) μmol/L), methionine ((37.0±26.9) μmol/L) and phenylalanine ((133.0±80.9)μmol/L) in Cit≥30 μmol/L group (n=18) were significantly different as compared with those in the other three groups, respectively (F=117.0, 58.5, 135.0, P<0.01). The levels of arginine ( (10.0±9.2) , (11.0±9.3) , (9.0±17.8) μmol/L), valine ( (119.0±29.8) , (107.6±14.1) , (102±68) μmol/L) and leucine ( (167.0±37.1) , (161.0±37.7) , (163.5±180.6) μmol/L) were not statistically significant among groups of Cit≥30 μmol/L(n=18), Cit21-<30 μmol/L(n=7) and Cit<21μmol/L(n=30,P>0.05), but they were significantly higher than those of the normal control group ((4±3), (78±21), (114.0±31.5) μmol/L, n=1 000), respectively(F=30.1, 23.0, 29.8, P<0.01). Alanine (Ala) ( (150±50) , (156.0±30.2), (168±105), (152±52) μmol/L) levels showed no significant difference (F=0.86, P>0.05) but the ratios of Ala/Cit (1.52±1.44, 6.82±1.56, 12.06±7.71, 19.42±6.27) decreased significantly among the four groups (F=69.0, P<0.05). The acylcarnitine levels showed no statistically significant results among the different groups (P>0.05). With Cit≥30 μmol/L and Ala/Cit<7.5 as cutoff values, the number of screen-positive cases reduced from 39 to 22 cases with no additional false negative case. With Cit≥21 μmol/L and Ala/Cit<7.5 as cutoff values the number of screen-positive cases increased to 117 cases with 1 additional true positive. Conclusions The profiles of blood amino acid in early neonates with NICCD present the increased levels of multiple amino acids including citrulline, tyrosine, methionine and phenylalanine, and decreased ratio of Ala/Cit. Taking citrulline and ratio of Ala/Cit as screening markers can improve the positive predictive value appropriately. The limited sensitivity of NICCD newborn screening may be related to early blood sampling time.

Objective To explore the application value of chromosomal microarray analysis(CMA)technolo-gy in children with abnormal development at the endocrine clinic,and to summarize the data of diagnosis and treatment. Methods A retrospective analysis of 15 children with abnormal development was performed at the endocrinology clinic of Guangzhou Women and Childrenˊs Medical Center from January 2015 to December 2017. The whole genome CMA was applied according to the standard operation procedure of CytoScan 750 arrays of Affymetrix,USA. The results were analyzed by chromosome analysis suite( CHAS)software and related bioinformatics methods. Results The report on CMA showed that the genomes of 15 children had the pathogenic copy number variation(CNVs)or variants of uncer-tain significance. The chromosomal abnormalities were consistent with the clinical manifestations of all children. There were deletions in 14 cases and duplications in 3 cases. Among the 15 cases,loss of heterozygosity was found in 2 cases, uniparental disomy in 1 case,trisomy in 2 cases,Turner syndrome in 2 cases,Smith-Magenis syndrome in 1 case,and wolf Hirschhorn syndrome in 1 case. Only 2 of 15 children were diagnosed as chromosomal abnormalities by routine kar-yotype analysis. Conclusions The whole genome high resolution CMA can significantly improve the rate of diagnosis in children with abnormal development at endocrinology clinic,and is worthy of recommendation.

OBJECTIVE:To investigate the effects of Qigong pills decoction combined with chemical drugs on hormone levels of polycystic ovarian syndrome(PCOS)patients complicated with infertility. METHODS:A total of 98 PCOS patients complicated with infertility selected from gynaecology department of our hospital during Jul. 2015-Jun. 2016 were divided into observation group and control group according to random number table,with 49 cases in each group. Control group was given Clomiphene ci-trate tablet 50 mg orally,qd,for consecutive 5 d;B-ultrasonography for follicle development was monitored at the 11th day of menstruation,and then they were given Chorionic Gonadotropin for injection 2000 U intramuscularly,qod,if there was not domi-nant follicle >10 mm till the diameter of follicle reached 18 mm. Observation group was additionally given one dose of Qigong pills decoction,every day 400 mL after decocted with water,each 200 mL for morning and night,on the basis of control group. After 3 months of treatment,the levels of FSH,LH,PRL,E2,T,DHEA-S and SHBG,the endometrial thickness,the number of mature follicle and pregnancy rate were compared between 2 groups. RESULTS:Before treatment,there was no statistical signifi-cance in the levels of FSH,LH,PRL,E2,T,DHEA-S or SHBG between 2 groups(P>0.05). 3 months after treatment,the lev-els of PRL,T and DHEA-S in 2 groups were decreased and E2,SHBG were increased significantly;the levels of FSH,PRL,T and DHEA-S in observation group were significantly lower than control group,while the levels of E2 and SHBG were significantly higher than control group,with statistical significance (P<0.05). Endometrial thickness of observation group were significantly higher than that of control group;pregnancy rate (18.37%) was significantly higher than control group (4.08%),with statistical significance (P<0.05). CONCLUSIONS:Qigong pills decoction combined with chemical drugs can help to improve pregnancy rate and regulate serum hormone level in the treatment of PCOS complicated with infertility.

To explore the effects and mechanism of dexamethasone and ambroxol on expression of surfactant protein (SP)-B mRNA and thyroid transcription factor (TTF)-1 in premature rat lung. Methods Sixteen pregnant Sprague-Dawley rats were randomly divided into four equal groups: two doses of dexamethasone (0.2 mg/kg injected intramuscularly on Day 17 and 18 of pregnancy respectively);single dose of dexamethasone (0.2 mg/kg injected intramuscularly on Day 18 of pregnancy);ambroxol group (100 mg/kg injected intraperitoneally on Day 16, 17 and 18 of pregnancy respectively); and control group (normal saline injected intraperitoneally on Day 16, 17 and 18 of pregnancy respectively). There were four pregnant rats in each group. All of the fetal rats were taken out on Day 19 of pregnancy as the preterm birth model, and 20 fetal rats from each group were randomly selected. The ratio of body weight to fetal lung weight of newborn rats was calculated. Changes in lung morphology were observed under light microscopy and the ratio of alveoli surface area to alveolar septae surface area was calculated. Expression of TTF-1 protein was determined by immunohistochemistry. Expression of SP-B mRNA was detected by reverse transcriptase-polymerase chain reaction. One-way analysis of variance, Student-Newman-Keuls method and Pearson correlation analysis were applied as statistical methods. Results (1) The ratio of body weight to fetal lung weight was (6.5±0.6), (7.9±0.8), (9.5±0.8) and (9.5±0.9) mg/g in two doses of dexamethasone group, one dose of dexamethasone group, ambroxol group and control group respectively (F=67.69,P<0.01). The ratio of two doses and one dose of dexamethasone group was lower than that of control group (q=17.143 and 9.143, all P<0.01) and ambroxol group (q=17.143 and 9.143, all P<0.01). The ratio of two doses dexamethasone group was lower than that of one dose dexamethasone group (q=8.000, P<0.01). (2) The ratio of alveoli surface area to alveolar septae surface area was 2.19±0.15, 1.70±0.18, 1.67±0.13 and 1.08±0.12 in two doses of dexamethasone group, one dose of dexamethasone group, ambroxol group and control group respectively (F=190.85, P<0.01). The ratio of two doses of dexamethasone group, one dose of dexamethasone group and ambroxol group were higher than that of the control group (q=33.639, 18.788 and 17.879, all P<0.01). The ratio of two doses dexamethasone group was higher than that of one dose dexamethasone group (q=14.848, P<0.01). (3) Expression of TTF-1 protein was 0.311±0.018, 0.224±0.019, 0.196±0.013 and 0.191±0.018 in two doses of dexamethasone group, one dose of dexamethasone group, ambroxol group and control group respectively (F=211.69,P<0.01). TTF-1 protein expression of two doses and one dose of dexamethasone group were higher than that of control group (q=30.000 and 8.250, all P<0.01) and ambroxol group (q=28.750 and 7.000, all P<0.01). TTF-1 protein expression of two doses dexamethasone group was higher than that of one dose dexamethasone group (q=21.750, P<0.01). (4) Expression of SP-B mRNA was 1.25±0.13, 1.15±0.12, 1.10±0.10 and 1.01±0.12 in two doses of dexamethasone group, one dose of dexamethasone group, ambroxol group and control group respectively (F=14.48, P<0.01). SP-B mRNA expression of two doses of dexamethasone group, one dose of dexamethasone group and ambroxol group were higher than that of control group (q=9.231, 5.385 and 3.462, all P<0.01). SP-B mRNA expression of two doses of dexamethasone group was higher than that of ambroxol group (q=5.769, P<0.01) and one dose of dexamethasone group (q=3.846, P<0.01). (5)TTF-1 expression in two doses of dexamethasone group, one dose of dexamethasone group and control groups was positively correlated with SP-B mRNA expression (r=0.512, 0.597 and 0.449, respectively, all P<0.05). Conclusions Ambroxol can accelerate the maturation of fetal lung with minimal adverse effects on fetal lung weight. Ambroxol might be an alternative to dexamethasone to prevent neonatal respiratory distress syndrome.

Objective By making models of premature animal,explores the effects of dexamethasone on the brain development of premature rats and its mechanisms.Methods Eighteen SD rats were randomly divided into high-dexamethasone(H-Dex) group,low-dexamethasone (L-Dex) group and normal saline(NS) control group,with 6 rats in each group.The pregnant rats in L-Dex group were injected with dexamethasone [0.1 mg/(kg·d)] from 16 to 18 days of pregnancy,while the pregnant rats in H-Dex group were injected with dexamethasone [0.5 mg/(kg· d)] ; the pregnant rats in NS control group were injected with 0.9％ NaCl of the same volume.All of the fetal rats were received after administrating caesarean operation on the day 19 of pregnancy.Rats were sacrificed at the directed time and brain tissue was prepared.Histological feature and the water content of the brains were observed.Level of apoptosis was measured by flow cytometry.The expressions of myelin basic protein (MBP) and interleukin(IL)-1β in brain tissue homogenate were detected by ELISA.Results (1) The brain water contents of rats in H-Dex group,L-Dex group and NS control group were (85.94 ± 0.54) ％,(86.08 ± 1.01) ％,(86.94 ± 0.82) ％.Compared with NS control group,the water contents of Dex group were lower (P ＜ 0.05).(2) Glial cells of brain cortex in L-Dex group and H-Dex group were more mature than in NS control group,and the changes in H-Dex group was more significant.(3) The expressions of MBP in brain tissue of H-Dex group,L-Dex group and NS control group were (5.73 ± 1.06) μg/mg,(5.46 ±0.77) μg/mg and (2.42 ±0.52) μg/mg.Compared with NS control group,Dex group was higher(P ＜0.05).While the expressions of IL-1β in brain tissue of H-Dex group,L-Dex group and NS control group were (249.05 ± 11.29) pg/g,(257.47 ± 9.33) and (292.66 ± 21.51) pg/g.Compared with NS control group,Dex group was lower(P ＜ 0.05).There was no significant difference between H-Dex group and L-Dex group(P ＞ 0.05).(4) The level of apoptosis in H-Dex group,L-Dex group and NS control group were (18.07 ± 1.63) ％,(6.88 ± 0.47) ％ and (2.00 ± 0.32) ％.Compared with NS control group,the level of apoptosis in Dex group was higher(P ＜0.05),and H-Dex group was higher than that in L-Dex group.Conclusion (1) Using dexamethasone prophylactic could promote the development of glial cells,reduce the water content,increase the expressions of MBP,and decrease the expressions of IL-1β in brain tissues.It indicates that dexamethasone may play a major role in maturation of fetal brain.(2) Using dexamethasone prophylactic could increase the amounts of the apoptosis cells,and this effect is dose-dependent.It indicates that dexamethasone may have a negative effect on the fetal brain and suggestes that using dexamethasone in premature infant should be cautious,and if it has to,using a lower dose.

Objective To evaluate the efficacy and safety of midodrine hydrochloride in the treatment of intradialysis hypotension (IDH)in maintenance hemodialysis (MHD)patients.Methods One hundred and tburteen MHD patients from 8 dialysis centers with IDH were enrolled in the study.These patients took orMly midodfine for 4～6 weeks.Midodrine(2.5～10 mg)was given 15～30 minutes after the beginning of hemodialysis,and another 2.5～10 mg was used during hemodialysis if systolic blood pressure(SBP)increased less than 20 mm Hg.The total usage of each dialysis session was not more than 20 mg.The pre-,intra-,post-hemodialysis blood pressure and heart rate,the pre-and post-hemodialysis body weight,the uhrafiitrated volume of each dialysis,the pre-and post-treatment liver and renal function and electrocardiogram were measured and recorded.The symptoms of IDH were observed. Results Compared to those before treatment with midodrine hydrochloride,the minimum intradialysis SBP and heart rate at that time,the post-dialysis SBP and heart rate,and total uhrafiitrated volume changed significanlly (P＜0.01).The total effective rate was 84.2%.And the symptoms of IDH were improved significantly (P＜0.01).The side effects were observed in only 2 patients.Conclusion Midodrine iS safe and effective for the treatment of IDH.