OBJECTIVES: To investigate the molecular epidemiology of children with phenylketonuria (PKU) in Gansu, China, providing foundational data for intervention strategies. METHODS: A retrospective analysis was conducted on 1 159 PKU families who attended Gansu Provincial Maternity and Child Care Hospital from January 2012 to December 2024. Sanger sequencing, multiplex ligation-dependent probe amplification, whole exome sequencing, and deep intronic variant analysis were used to analyze the PAH gene. RESULTS: For the 1 159 children with PKU, 2 295 variants were identified in 2 318 alleles, resulting in a detection rate of 99.01%. The detection rates were 100% (914/914) in 457 classic PKU families, 99.45% (907/912) in 456 mild PKU families, and 96.34% (474/492) in 246 mild hyperphenylalaninemia families. The 2 295 variants detected comprised 208 distinct mutation types, among which c.728G>A (14.95%, 343/2 295) had the highest frequency, followed by c.611A>G (4.88%, 112/2 295) and c.721C>T (4.79%, 110/2 295). The cumulative frequency of the top 23 hotspot variants reached 70.28% (1 613/2 295), and most variant alleles were detected in exon 7 (29.19%, 670/2 295). CONCLUSIONS Deep intronic variant analysis of the PAH gene can improve the genetic diagnostic rate of PKU. The development of targeted detection kits for PAH hotspot variants may enable precision screening programs and enhance preventive strategies for PKU.
Humans ; Phenylketonurias/epidemiology* ; Female ; Male ; Retrospective Studies ; Phenylalanine Hydroxylase/genetics* ; Mutation ; Child, Preschool ; China/epidemiology* ; Child ; Infant

Objective: To investigate the differences and diversity changes in gut microbiota between children and adolescents with constipation and diarrhea, and healthy individuals, and to explore the correlation between changes in stool consistency and gut microbiota, in order to provide a scientific reference for the research on intestinal microecology among children and adolescents. Methods: From October 2021 to March 2022, a total of 42 children and adolescents with constipation and 37 with diarrhea from a tertiary hospital in Hangzhou City, and 43 healthy individuals from 3 primary and secondary schools were included in this study. Fecal samples of children and adolescents were collected and then stool genomic DNA was extracted for 16S rRNA gene high throughput sequencing, and the sequencing results were analyzed. In the analysis of alpha diversity, the Kruskal-Wallis rank sum test was used to compare the differences between the three groups, and the FDR multiple testing correction was used for pairwise comparisons. In the analysis of beta diversity, the Adonis test was used to compare the overall differences between the three groups, and the ANOSIM test was used for pairwise comparisons. In the LEfSe analysis, the LDA scores obtained through LDA analysis (linear regression analysis). Results: Alpha diversity analysis showed that there were statistically significant differences in the Shannon index (4.01, 3.81, 4.19) and Simpson index (0.05, 0.06, 0.04) between the diarrhea group, constipation group, and healthy group ( H=6.05, 6.35, P <0.1). Further pairwise comparison showed that the Shannon index and Simpson index of the healthy group were higher than those of the constipation group ( P <0.1). Beta diversity analysis showed that the impact of grouping factors on inter group differences was statistically significant ( R 2=0.045, P <0.1). Community composition analysis showed that there were 234 species in total among the three groups, and 36 unique species in the healthy group, 36 species in the diarrhea group, and 48 species in the constipation group. Species difference analysis showed significant differences in species composition at the genus level among the three groups ( H=0.000 05, 0.000 16, 0.000 20, 0.000 21, 0.000 53, 0.001 39, P <0.1), including Lachnospiraceae of Firmicutes phylum, Eubacterium hallii, Veillonellaceae, Qscillospiraceae, Butyricicoccaceae and Staphylococcaceae, respectively. KEGG abundance statistics and COG functional analysis showed that there were no significant differences in gene expression abundance of the same function among the three groups ( P >0.1). Conclusions The different stool consistency of children and adolescents is related to changes in gut microbiota composition. Compared to the healthy group, children with constipation or diarrhea have disrupted gut microbiota balance, with a shift in dominant bacteria and a higher abundance of opportunistic pathogens.

OBJECTIVE:To provide reference for pharmacists to participate in the management of chronic disease. METHODS:A total of 259 patients with chronic airway disease [included asthma and chronic obstructive pulmonary disease (COPD)] met the inclusion criteria were selected from our hospital and 5 community health care centers of medical consortium. These patients received medication safety assessment management,which was led by clinical pharmacists of our hospital with the participation of community pharmacists,including medication safety comprehensive evaluation and risk classification management, follow-up and medication guidance, integrated prescriptions checking, establishment of shared database. 1 years after the implementation,the effectiveness were evaluated by score the relatived indicators in related groups. RESULTS:After a year of the management mode practice,compared with before intervention,the patients'safety medication cognitive ability score in high-risk and low-risk group increased from(4.49±1.26)and(7.31±1.01)to(5.40±1.56)and(7.44±0.91);medication adherence score increased from(4.96±1.21)and(7.08±1.24)to(6.66±1.08)and(7.38±0.98);ACT score from asthma patients increased from (16.15±2.58)and(21.15±1.03)to(16.80±2.57)and(21.64±1.55);CAT score from COPD patients decreased from(25.51± 4.07) and (14.90 ± 3.95) to (24.20 ± 3.96) and (13.80 ± 4.08);the rate of irrational prescription effective identification and intervention by pharmacists increased from 3.6％ and 1.4％ to 9.4％ and 7.6％,respectively. All the differences above were statistically significant (P<0.05). CONCLUSIONS:The participation of pharmacists in long-term medication safety assessment management for chronic airway disease patients can improve patients'safety medication cognitive ability,medication adherence, disease control and the pharmacists'ability of irrational drug use identification and intervention.

Objective To evaluate the effect for stroke(cerebral infarction)at acute stage intervened by traditional Chinese medicine(TCM) clinical pathway. Methods A retrospective analysis was carried out in the patients before and after intervention by TCM clinical pathway during the year of 2013-2016. The outcomes included the entrance rate,completion percentage,aberration rate,severity,hospitalization time,and hospitalization fee as well as functional prognosis 30 d after discharge. Results A total of 792 cases was included,241 cases admitted in the year of 2013, 191 in the year of 2014, 135 in the year of 2015, and 225 in the year of 2016. In the year of 2014-2016,the entrance rate,completion percentage,hospitalization time,hospitalization fee,critical case percentage,and gender constituent ratio were steadily(P > 0.05). Modified Rankin Scale(mRS)for short-term functional prognosis mRS 0 ~ 2 percentage and aberration rate in the year of 2016 were superior to those in the year of 2014 and 2015(P < 0.05). Functional improvement on discharge day 30 in the year of 2014-2016 was superior to that in the year of 2013(P < 0.05). The hospitalization time and hospitalization fee in the year of 2014-2016 were a liffle higher than those in the year of 2013(P < 0.05), showing no obvious increase. Conclusion The application of TCM clinical pathway can make the diagnosis and treatment procedure of stroke at acute stage standardized,enhance the therapeutic effect,achieve or even exceed the goal of manage scheme.

Objective To investigate the clinical efficacy of sertraline combined with famciclovir in treatment of herpes zoster neuralgia . Methods Ninety -four patients with herpes zoster neuralgia were randomly divided into treatment group and control group , 47 cases in each group. Two groups of patients were performed conventional nutritional and physical therapy . Control group was treated with oral administration of famciclovir 0.25 g, tid, treatment group added sertraline 50 mg? d-1 on the basis of control group . The course of treatment was 4 weeks. The visual analogues scale ( VAS ) , clinical efficacy , quality of life and the adverse drug reactions were compared between the two groups.Results In the treatment group , the VAS in 1 and 4 weeks were ( 5.13 ±1.26 ), (3.27 ±1.02), significantly decreased than before treatment ( P<0.05 ) , and significantly lower than control group, which were(6.17 ±1.57) and (4.87 ±1.13).The total effective rate in treatment group ( 93.62%) was higher than control group ( 74.47%, P <0.05 ) .The quality of life in two groups were significantly increased after treatment ( P<0.05 ) , and treatment group significantly higher control group (P<0.05).No serious adverse drug reaction was found in two groups.Conclusion The clinical efficacy of seculin combined with famciclovir in the treatment of herpes zoster neuralgia is significant.

Objective To observe and study the clinical efficacy of laparoscopy combined with hysteroscopy in treatment of obstruction of fallopian tube.Methods 72 cases of patients who were treated for fallopian tubal obstruction were selected as the research object,and they were randomly by simple randomization divided into two groups,the observation group and the control group,with 36 cases in each group.The hysteroscopic treatment was used in the control group,hysteroscopy combined with laparoscopy were treated in the observation group.Then operation effect and 1 years after surgery in pregnancy of the two groups were compared.Results In the control group,70 fallo-pian tube had 56 successful recanalization with the recanalization rate of 80%.At 3 months after operation it blocked 8,restenosis rate was 14.2%.In the observation group,68 fallopian tube had 62 successful recanalization with the recanalization rate of 91.1%,and at 3 months after operation it blocked 10,restenosis rate 16.2%.The recanalization rate in the observation group was significantly higher than that of the hysteroscopy wire group(χ2 =6.231,P <0.05). The restenosis rate was no significant difference between the two groups(P >0.05).The control group of 14 cases with intrauterine pregnancy,the pregnancy rate was 38.9%,1 cases of tubal pregnancy,the pregnancy rate was 2.7%.24 cases of pregnancy with intrauterine in the observation group,the pregnancy rate was 66.7%,2 cases of tubal pregnancy,the pregnancy rate was 5.4%.The intrauterine pregnancy rate in the observation group was signifi-cantly higher than that of the hysteroscopy group,there was significant difference(χ2 =5.568,P <0.05),fallopian tube pregnancy rate was no significant difference between the two groups(P >0.05 ).Conclusion Hysteroscopy combined with laparoscopy in treatment of fallopian tubal obstruction can make up for the deficiency of hysteroscopic treatment alone,it can greatly improve the fallopian tube recanalization rate and pregnancy rate after operation,it is worthy for clinical application.

Objective To study the effect ofdifferent doses of dexmedeto-midine to pain , stress and immune function in elderly patients with lapa-roscopic surgery Methods One hundred twenty -six patients of laparo-scopic surgery were randomly divided into low -dose group ( 42 cases ) , middle dose group (42 cases), and high-dose group (42 cases).Be-fore anesthesia 15 min, the low dose group , middle dose group and high dose groups were intravenouslygiven 0.2 , 0.4 , 0.6 μg · kg -1 · h-1 of dexmedetomidine ,respectively.Diastolic bllod pressure ( DBP) , systolic blood pressure ( SBP) , blood sugar ( Glu) , heart rate ( HR) to the hori-zontal and CD4, CD8, CD4/CD8 levels of before and after surgery 3, 8, 24 h were compared.Results The pain scores ,DBP, SBP, Glu and HR levels of before and after surgery 24 h between three groups were not sig-nificant differences ( P>0.05 ).After surgery 3 h and 8 h, pain scores of middle high dose groups were significantly higher than that of low -dose group ( P <0.05 ) and their DBP, SBP, Glu, HR were significantly better than that of low-dose group(P<0.05).Before and after surgery, CD4 levels in three groups of patients did not change significantly.Before and 3 h after surgery, CD8, CD4/CD8 levels in the three groups were not significant difference ( P>0.05 ).After surgery 8 h and 24 h, CD8 levels of high-dose group was significantly lower than of the low -dose group , and the CD4/CD8 ratio was significantly better than low -dose group(P<0.05).Conclusion The use of 0.4, 0.6 μg· kg -1 · h-1 dexmedetomidine scheduled for laparo-scopic surgery in elderly patients have a more significant analgesic effect , can reduce stress and improve immune func-tion in patients , it is worth widely used.

Transforming growth factor-β1 (TGF-β1)-activated phosphoinositide-3-kinase (PI3K)-protein kinase B (PKB/Akt) pathway is intimately related to the development of diabetic nephropathy (DN), which is negatively regulated by phosphatase and tensin homolog deleted on chromosome ten (PTEN). The present study was to investigate the expression of PTEN in the renal tissue of diabetic mellitus (DM) rats and explore its possible effect on development of DN. Sixteen Sprague-Dawley rats were divided into normal control group (n = 8) and diabetic group (n = 8) at random. Streptozotocin injection was used to establish diabetic model. After 12 weeks, the rats were sacrificed to detect relative biochemical parameters and renal index, and to observe the changes of pathomorphology by HE staining as well. In addition, immunohistochemistry staining and Western blotting were employed to detect the protein expression of PTEN, TGF-β1, PI3Kp110α, Akt1, p-Akt1 (Ser(473)), fibronectin (FN) and Collagen IV, respectively. Furthermore, the expression of PTEN mRNA was also examined by RT-PCR. The results indicated that the levels of blood glucose, serum creatinine and urine protein (24 h) were increased remarkably in the diabetic group (P < 0.05) compared with those in the control group. Compared with those in the control group, the protein expressions of TGF-β1, PI3Kp110α, Akt1 in renal tubular epithelium and the expressions of FN and CollagenIV in renal interstitium were increased in the diabetic group (P < 0.05). The expression of PTEN in the diabetic group was significantly reduced than that in the control group (P < 0.05), and the expression of p-Akt1 (Ser(473)) increased remarkably in the diabetic group which had the similar trend to Akt1 (P < 0.05). PTEN mainly located in renal tubular epithelial cells. The expression of PTEN had negative correlation to that of p-Akt1 (Ser(473)). Compared with that in the control group, the expression of PTEN mRNA was decreased remarkably in the diabetic group (P < 0.05). The data suggest that the down-regulation of PTEN in renal tissue of DM rats may promote the PI3K-PKB/Akt pathway over-activated by TGF-β1, which facilitates the initiation and development of DN.
Animals ; Diabetes Mellitus, Experimental ; complications ; Diabetic Nephropathies ; metabolism ; Down-Regulation ; Kidney ; metabolism ; Male ; PTEN Phosphohydrolase ; genetics ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVEThe relationship between latent adenovirus infection and airway inflammation have not been well documented. The aim of this study is to illustrate the roles of adenovirus E1A protein on the level of glutathione (GSH) in response to oxidative stress and the effect of the oxidant/antioxidant imbalance upon the transactivation of NF-kappaB triggered by E1A protein.

METHODSRat alveolar epithelial cell stably expressing adenoviral E1A or control plasmid were developed. For isolation of nuclear extracts, 5 x 10(5) cells were plated and grown overnight in 60 mm dishes. Experiments were repeated three times. The cell model of stably expressing adenoviral E1A was stimulated by H2O2. The level of GSH were measured. E1A positive clone was stimulated by LPS or TNF-alpha and treated with L-Buthionine-sulfoximine (BSO). The expression of NF-kappaB was measured by Western blot. Differences between groups were assessed for significance by Student' t test; multiple comparisons by the one-way ANOVA.

RESULTSThere is no difference of GSH level without stimulation between E1A-positive clones and E1A-negative clones. For E1A-positive clones, the level of GSH did not increase in response to H2O2 as E1A-negative clones. The quantitation by densitometry of the NF-kappaB expression in E1A-positive clones were (79.3 +/- 4.6), (80.3 +/- 3.8) respectively without treatment and were (81.8 +/- 3.9) - (89.9 +/- 1.6) and (94.1 +/- 1.9) - (99.8 +/- 1.6) respectively under LPS or TNF-alpha stimulation, which were significantly higher than that of the control group (68.3 +/- 3.8), (69.4 +/- 4.3) respectively without stimulation and (70.1 +/- 2.8) - (80.8 +/- 3.6), (73.4 +/- 4.9) - (83.2 +/- 6.7) respectively under stimulation. The quantitation by densitometry of the NF-kappaB expression in E1A-negative clones were (1.25 +/- 0.18) and (1.69 +/- 0.19) respectively under LPS and TNF-alpha-stimulation and (1.22 +/- 0.16) and (1.75 +/- 0.13) respectively upon treatment for LPS and TNF-alpha with BSO preincubation. There did not show difference upon treatment with LPS or TNF-alpha with or without BSO in E1A-negative cell clone. The quantitation by densitometry of the NF-kappaB expression in E1A-positive clone were (1.75 +/- 0.10) and (2.26 +/- 0.21) respectively upon treatment for LPS and TNF-alpha with BSO preincubation which were significantly higher than that of LPS or TNF-alpha-stimulation alone (1.35 +/- 0.12), (1.80 +/- 0.14) respectively.

CONCLUSIONThese results indicate that E1A protein decreased GSH levels in oxidant stress and upregulated NF-kappaB transcription activity. The oxidant/antioxidant imbalance in rat alveolar epithelial cells enhances E1A-modulated transcriptional activation of NF-kappaB. The mechanism underlying transactivation of NF-kappaB involved by E1A may be related to oxidative stress.

Adenovirus E1A Proteins ; genetics ; metabolism ; Animals ; Cell Line ; Epithelial Cells ; metabolism ; NF-kappa B ; genetics ; metabolism ; Oxidative Stress ; Pulmonary Alveoli ; cytology ; Rats ; Transcriptional Activation