Objective To study the effect of Lishukang capsule on learning and memory impairment in mice with high altitude hypoxia based on Keap1/Nrf2/HO-1 signal pathway. Methods Sixty male Balb/C mice were randomly divided into normal control group, hypoxia model group, Rhodiola capsule group (400 mg/kg), low, medium and high dose groups of Lishukang capsule (400、 600、 800 mg/kg), with 10 mice in each group. The normal control group was fed at the local altitude (1 500 m) after 7 days of intragastric administration in each group, and the rest groups were fed at the low pressure and hypoxia animal experimental cabin to simulate the altitude of 7500 m for hypoxia for 3 days. During this period, the normal control group and the hypoxia model group were given normal saline once a day, and 1 hour after the last administration, the eight arm maze was used to test the spatial memory ability of mice under simulated high altitude hypoxia; HE staining was used to observe the morphological changes of hippocampus in mice; Western blot was used to detect the changes of protein content of Keap1/Nrf2/HO-1 signal pathway and apoptosis related protein in hippocampus of mice. Results Compared with the normal control group, the spatial memory ability of mice in the hypoxia model group was significantly impaired (P<0.01); HE staining showed that hippocampal neurons in mice were seriously injured; the content of brain tissue Keap1 protein and apoptosis related protein Bax and Caspase-3 increased (P<0.01); the content of Nrf2, HO-1 and apoptosis related protein Bcl-2 decreased （P<0.01）. Compared with the hypoxia model group, the error rate of mice in the high dose group of Lishukang capsule in the eight arm maze behavior experiment was significantly reduced (P<0.05, P<0.01); HE staining showed that the neurons were arranged orderly and the cell morphology was good; the content of Keap1 protein and apoptosis related protein Bax and Caspase-3 decreased (P<0.01); the content of Nrf2, HO-1 and apoptosis related protein Bcl-2 increased（P<0.01）. Conclusion High altitude hypoxia can lead to oxidative stress injury in mice and induce the expression of apoptosis related genes, thus aggravating the cognitive dysfunction of mice; Lishukang capsule can effectively improve the learning and memory impairment in mice caused by hypoxia, and its mechanism may be related to regulating the Keap1/Nrf2/HO-1 signal pathway and reducing apoptosis.

Objective:To investigate the clinical application value of whole brain computed tomography perfusion(CTP)imaging combined with computed tomography angiography(CTA)on head and neck in treating chronic ischemic cerebrovascular disease(ICVD)by superficial temporal artery-middle cerebral artery(STA-MCA)bypass.Methods:Fifty patients with suspected chronic ICVD who admitted to Chengdu Second People's Hospital and Deyang People's Hospital(29 cases from Chengdu Second People's Hospital and 21 cases from Deyang People's Hospital)from May 2020 to October 2024 were selected.All patients underwent whole-brain CTP examination and CTA examination on head and neck before the surgery,and then,the preoperative assessment efficacy of the combination of the two examination for patients with STA-MCA surgical indication was analyzed.The changes of the CTP parameters[cerebral blood volume(CBV),cerebral blood flow(CBF),mean transit time(MTT),time to drain(TTD),and time to maximum(Tmax)]between the affected side and the healthy side before surgery,and between them after surgery,and these parameter of affected side between before and after surgery were compared respectively.The course and the anastomosis conditions of the bypass vessels were analyzed through CTA images.The CTP parameters of patients with postoperative cerebral hyperperfusion syndrome(CHS)were statistically analyzed.Then,the relationship between the changes of cerebral perfusion values on the affected side after surgery and CHS,as well as the follow-up situation,were analyzed.Results:In 50 patients with suspected chronic ICVD,34 patients with surgical indication of STA-MCA bypass were screened out by CTA combined with CTP,who existed severely narrowed or occluded blood vessels accompanied by reduced CTP,and other 16 cases who did not meet surgical indication of STA-MCA bypass were excluded.In 34 patients with chronic ICVD,the preoperative CBF value of affected side was lower than that of healthy side,and the preoperative MTT,TTD and Tmax values of affected side were all higher than those of the healthy side,and the differences were statistically significant(t=6.648,6.030,10.638,14.063,P<0.05),while there was not significant difference in CBV value between two sides(P>0.05).In the affected side,the postoperative CBF value was higher than preoperative CBF value,and the postoperative MTT,TTD and Tmax values were lower than preoperative these indicators,and the differences were statistically significant(t=8.578,7.166,11.321,11.912,P<0.05).There was no significant difference in the CBV value between before and after surgery(P>0.05).In 34 patients with chronic ICVD,there was not significant difference in CTP parameters between the affected side and the healthy side in 28 patients with chronic ICVD after 6 patients with CHS were excluded(P>0.05).The results of CTA showed that the diameter size and course of superficial temporal artery of all patients were clearly displayed,and the bypass vessels of them were unblocked,and the anastomoses of them were well developed.All 34 patients with chronic ICVD successfully completed unilateral STA-MCA bypass surgery.After the surgery,based on the clinical symptoms and the CTP reexamination results of the patients,it was determined that 6 patients(17.7%)were CHS,whose appearances were the CBF value of affected side was(60.81±5.79)ml/100(g·min),which was significantly higher than that[(53.30±5.21)ml/100(g·min)]of the healthy side.The MTT value of affected side was(2.9±0.82)s,which was significantly lower than that of the healthy side(4.2±1.1)s,and the differences were statistically significant(t=2.358,2.341,P<0.05).In the postoperative follow-up of 2 to 17 months for patients,there was not dead case,and the clinical symptoms of the most of patients were improved significantly,and there was no serious adverse events.Conclusion:Whole-brain CTP combined with CTA examination on head and neck is conducive to accurately determine the degree of vascular stenosis and cerebral perfusion before surgery,and screen patients with chronic ICVD who occurred indications of STA-MCA surgery,and can also assess the patency of bypass vessels,the recovery degree of cerebral perfusion,and the possibility of occurring CHS after surgery,which has important clinical guiding value.

Objective:The greater tuberosity angle(GTA)and critical shoulder angle(CSA)are commonly referred to as radiographic markers which were used to described morphology of the greater tu-berosity and acromion respectively.At present,most international studies focus on the correlation be-tween the above two parameters and rotator cuff tears(RCTs),and their diagnostic value and risk assess-ment.This study attempts to find out the trend of GTA and CSA changes and risk threshold of RCTs,as well as the protective factors and risk factors.Methods:In this study,130 individuals from May 2019 to December 2020 were recruited.According to Southern California Orthopedic Institute(SCOI)classifica-tion,the individuals were divided into four groups retrospectively:Group A,negative control group;Group B,partial tears(articular side);Group C,partial tears(bursal side);Group D,full-thickness tears.GTA and CSA were measured respectively on true anteroposterior position X-ray of shoulder with arm in neutral rotation and performed by the same trained technician team in single-blind.The correla-tions between RCTs and relevant factors were analyzed.Results:According to the area under the receiver operating characteristic curve(AUC),GTA and CSA of RCTs(Groups B,C and D)were 0.736 and 0.673 with 95％confidence interval(CI),the cut-off value of GTA and CSA of RCTs were 70.5° and 39.5° respectively.Comparing with the control group,RCTs groups had significant statistical differences in age and body mass index(BMI)(P＜0.05),especially the full-thickness RCTs(Group D),which was older than Groups A,B and C(P＜0.05,cut-off value:56.5 years old)and shorter than Groups A and B(P＜0.05,cut-off value:1.58 m).Analyzed from scatter plot and regression analysis,there was no linear correlation between GTA and CSA.There were no significant differences in gender,dominant shoulders and smoking between the RCTs groups and the control group(P＞0.05).Conclusion:Larger GTA(＞70.5°)and CSA(＞39.5°)would be highly predictive in diagnosing RCTs without linear cor-relation,and GTA has a higher diagnostic value in contrast.Subacromial impingement and shoulder de-generation occurred before RCTs.Patients with age＞56.5 years and height＜1.58 m were more likely to develop disease of full-thickness RCTs and no statistic differences in weight and BMI.Gender,domi-nant shoulder and smoking were neither risk factors nor protective factors.

Objective:To explore the genetic testing outcomes of a fetal family with Thyroid secretion disorder type 5 (TDH5) and familial Neurofibromatosis type 1 (NF1), and to clarify the association between clinical manifestations and genetic variations.Methods:One case of a TDH5 combined with familiar NF1 fetus treated at Gansu Maternal and Child Health Hospital in January 2024 was selected as the research subject. The clinical and family history data of the fetus were collected by retrospective research method. 10-15 mL of fetal amniotic fluid, and 2-3 mL of peripheral blood from the parents, sister, and grandfather of the fetus were collected, and genomic DNA was extracted for trio whole-exome sequencing (trio-WES). The Sanger sequencing was utilized to validate candidate variants for family verification. According to the Standards and Guidelines for the Interpretation and Reporting of Sequence Variants of the American Society of Medical Genetics and Genomics (ACMG) (hereafter referred to as the ACMG guidelines), the pathogenicity of the detected variants was classified. This study has been approved by the Medical Ethics Committee of Gansu Maternal and Child Health Hospital [Ethics No.(2021)GSFY(65)].Results:The fetal ultrasound indicated the nuchal translucency (NT) thickening, and the thyroid function test results of the sister showed an increase in thyroid stimulating hormone and a decrease in free thyroid hormone. Simultaneously, there were cafe-au-lait macules of various sizes in multiple parts of the body of the sister, and the mother had a similar cafe-au-lait macules phenotype. The trio-WES results revealed that there was a c. 413dupA(p.Tyr138*) frameshift mutation in exon 4 and c. 573G>A(p.Trp191*) nonsense mutation in exon 5 of the fetal DUOXA2, which were inherited from the mother and father, respectively. In accordance with the ACMG guidelines, they were classified as pathogenic variant (PVS1+ PM2_Supporting+ PM3) and likely pathogenic variant (PVS1+ PM2_Supporting), respectively. And the nonsense mutation c. 6972C>A(p.Tyr2264*) was detected in exon 46 of the NF1 in the fetus, inherited from the mother. The genetic testing results of the first sister and proband in this case were consistent, and the DUOXA2 and NF1 of the second sister were both wild-type. According to the ACMG guidelines, c.6972C>A(p.Tyr2264*) was classified as pathogenic variant (PVS1+ PS4_Supporting+ PP4+ PM2_Supporting). Conclusion:The mutations in the DUOXA2 gene c. 413dupA(p.Tyr138*) and c. 573G>A(p.Trp191*), and the NF1 gene c. 6972C>A(p.Tyr2264*) might be the genetic causes of TDH5 combined with familiar NF1 in proband. The discovery of the DUOXA2 gene c. 573G>A(p.Trp191*) enriches the spectrum of pathogenic gene variations.

Objective To investigate the expression of forkhead box P3(Foxp3)in peripheral blood of children with bronchial asthma and its relationship with airway hyper-responsiveness and body allergy.Methods A total of 80 children with bronchial asthma treated in the Affiliated Hospital of Yangzhou University from July 2024 to June 2025 were selected as experimental group,and 47 health-y children with physical examinations in the same period were selected as control group.Airway hy-per-responsiveness indicators(respiratory resistance,bronchial stimulant reaction threshold,and re-sistance increase slope),body allergy indicators[serum immunoglobulin E(IgE),eosinophil cationic protein(ECP),and eosinophil count(EOS)],pulmonary function indicators[peak expiratory flow(PEF),forced expiratory volume in the first second(FEV,),forced vital capacity(FVC),and the ratio of forced expiratory volume in the first second to forced vital capacity(FEV1/FVC)],and the expression level of Foxp3 in peripheral blood were compared between the two groups.Pearson corre-lation analysis was used to explore the correlations of Foxp3 with airway hyper-responsiveness and body allergy.Results The respiratory resistance and resistance increase slope in the control group were significantly lower than those in the experimental group,while the reaction threshold was signif-icantly higher than that in the experimental group(P＜0.05).The levels of IgE,ECP and EOS in the experimental group were significantly higher than those in the control group(P＜0.05).The FEV1,FVC,PEF,and FEV1/FVC in the experimental group were significantly lower than those in the control group(P＜0.05).The expression level of Foxp3 in the experimental group was(3.04±0.22),which was significantly lower than(4.22±0.41)in the control group(P＜0.05).Foxp3 was significantly correlated with respiratory resistance(r=-0.700,P＜0.001),reaction thresh-old(r=0.704,P＜0.001),resistance increase slope(r=-0.842,P＜0.001),IgE(r=0.864,P＜0.001),ECP(r=-0.684,P＜0.001),and EOS(r=-0.854,P＜0.001).Con-clusion The decreased Foxp3 in peripheral blood in children with bronchial asthma is associated with aggravated airway hyper-responsiveness and body allergy,suggesting that Foxp3 may be in-volved in the pathogenesis of bronchial asthma.

OBJECTIVE: To explore the genetic testing outcomes of a fetal family with Thyroid dyshormonogenesis type 5 (TDH5) and familial Neurofibromatosis type 1 (NF1), and to clarify the association between clinical manifestations and genetic variations. METHODS: One case of a TDH5 combined with familiar NF1 fetus treated at Gansu Maternal and Child Health Hospital in January 2024 was selected as the research subject. The clinical and family history data of the fetus were collected by retrospective research method. 10-15 mL of fetal amniotic fluid, and 2-3 mL of peripheral blood from the parents, sister, and grandfather of the fetus were collected, and genomic DNA was extracted for trio whole-exome sequencing (trio-WES). The Sanger sequencing was utilized to validate candidate variants for family verification. According to the Standards and Guidelines for the Interpretation and Reporting of Sequence Variants of the American Society of Medical Genetics and Genomics (ACMG) (hereafter referred to as the ACMG guidelines), the pathogenicity of the detected variants was classified. This study has been approved by the Medical Ethics Committee of Gansu Maternal and Child Health Hospital [Ethics No.（2021）GSFY（65）]. RESULTS: The fetal ultrasound indicated the nuchal translucency (NT) thickening, and the thyroid function test results of the sister showed an increase in thyroid stimulating hormone and a decrease in free thyroid hormone. Simultaneously, there were cafe-au-lait macules of various sizes in multiple parts of the body of the sister, and the mother had a similar cafe-au-lait macules phenotype. The trio-WES results revealed that there was a c.413dupA (p.Tyr138*) frameshift mutation in exon4 and c.573G>A (p.Trp191*) nonsense mutation in exon5 of the fetal DUOXA2, which were inherited from the mother and father, respectively. In accordance with the ACMG guidelines, they were classified as pathogenic variant (PVS1+PM2_Supporting+PM3) and likely pathogenic variant (PVS1+PM2_Supporting), respectively. And the nonsense mutation c.6972C>A (p.Tyr2264*) was detected in exon46 of the NF1 in the fetus, inherited from the mother maternal grandfather. The genetic testing results of the first sister and proband in this case were consistent, and the DUOXA2 and NF1 of the second sister were both wild-type. According to the ACMG guidelines, c.6972C>A (p.Tyr2264 *) was classified as pathogenic variant (PVS1+PS4_Supporting+PP4+PM2_Supporting). CONCLUSION The mutations in the DUOXA2 gene c.413dupA (p.Tyr138*) and c.573G>A (p.Trp191*), and the NF1 gene c.6972C>A (p.Tyr2264*) might be the genetic causes of TDH5 combined with familiar NF1 in proband. The discovery of the DUOXA2 gene c.573G>A (p.Trp191*) enriches the spectrum of pathogenic gene variations.
Humans ; Female ; Pedigree ; Pregnancy ; Neurofibromatosis 1/complications* ; Male ; Genetic Testing ; Adult ; Thyroid Dysgenesis/genetics* ; Fetus ; Exome Sequencing ; Mutation

BACKGROUND:miRNAs expression has been reported to be associated with hepatic and renal fibrosis,and dermal fibrogenesis. Moreover,a targeted regulatory relationship between miR-192-5p and epidermal regulators has been demonstrated in gouty arthritis.OBJECTIVE:To investigate the expression and regulatory role of miR-192-5p in hypertrophic scar and to verify whether there is a targeted regulatory relationship between miR-192-5p and epidermal regulators. METHODS:(1) Six cases of hypertrophic scar tissue and six cases of normal skin tissue were collected from the First Affiliated Hospital of Xinjiang Medical University. And miR-192-5p and epidermal regulator mRNA expression were detected by qRT-PCR. (2) The primary hypertrophic scar fibroblasts were obtained using tissue explant method and cultured to 3-6 generations for subsequent experiments. There were three groups in the experiment:negative control group,miR-192-5p mimic group and miR-192-5p inhibitor group. The latter two groups were transfected with the corresponding sequences. Cell proliferation viability was detected by the cell counting kit-8 assay and EdU kit;and the migration ability was detected by the cell scratch test. Cell apoptosis was detected by flow cytometry. The gene and protein expressions of epidermal regulator,type Ⅰ collagen,type Ⅲ collagen and α-smooth muscle actin were detected by qRT-PCR and western blot,respectively. miR-192-5p targets were predicted by a bioinformatics website,and target binding was validated by dual luciferase assay. RESULTS AND CONCLUSION:(1) Compared with normal skin tissues and their fibroblasts,miR-192-5p and epidermal regulator were highly expressed in hypertrophic scar and hypertrophic scar fibroblasts (P<0.05 or P<0.01). (2) After overexpression of miR-192-5p,cell proliferation was enhanced (P<0.05) and EdU positive cell rate increased (P<0.01) when compared with the negative control group;after inhibition of miR-192-5p,cell viability (P<0.05) and EdU positive rate decreased (P<0.05). (3) At 24 hours after overexpression of miR-192-5p,compared with the negative control group,the area between cell scratches and apoptosis rate decreased in the miR-192-5p mimic group (P<0.05) but increased in the miR-192-5p inhibitor group (P<0.01). (4) At 48 hours after transfection,the mRNA and protein levels of epidermal regulator were significantly decreased in the miR-192-5p mimic group,while the mRNA and protein levels of type Ⅰ collagen,type Ⅲ collagen and α-smooth muscle actin were significantly increased (P<0.05 or P<0.01). The miR-192-5p inhibitor group showed opposite changes in the above four indicators (P<0.05 or P<0.01). (5) The Targetscan website predicted that epidermal regulator had a potential binding site for miR-192-5p. (6) Dual luciferase assays showed that miR-192-5p could bind to epidermal regulator in a targeted manner. To conclude,overexpression of miR-192-5p can decrease the expression of epidermal regulator,and the two may be negatively regulated,suggesting that regulation of epidermal regulator may play a role in inhibiting the proliferation of hypertrophic scar fibroblasts.

Objective To analyze the clinical manifestations and genetic etiology of three families with hereditary spastic paraplegia(HSP).Methods Gene analysis was performed on patients of the three HSP families from the Gansu Provincial Maternity and Child-care Hospital.Results The proband of family 1 was autosomal recessive spastic paraplegia type 35 caused by homozygous variant c.159_176delGGCGGGCCAGGACATCAG(p.Arg53_Ser59delinsSer)in FA2H.The proband in family 2 was autosomal recessive spastic paraplegia type 47 caused by homozygous variant c.1399G>T(p.Glu467Ter)in AP4B1,and the proband in family 3 was autosomal recessive spastic paraplegia type 11 caused by homozygous variation c.7023C>G(p.Tyr2341Ter)in SPG11.Among them,the variant c.1399G>T(p.Glu467Ter)of AP4B1 is a novel variant,that has not been reported before,according to the ACMG guidelines,the pathogenicity of this variant is pathogenic.Conclusion This study has expanded the variant spectrum of AP4B1 which provides basic data to improve clinical understanding and diagnostic capabilities of HSP patients.

Objective:The greater tuberosity angle(GTA)and critical shoulder angle(CSA)are commonly referred to as radiographic markers which were used to described morphology of the greater tu-berosity and acromion respectively.At present,most international studies focus on the correlation be-tween the above two parameters and rotator cuff tears(RCTs),and their diagnostic value and risk assess-ment.This study attempts to find out the trend of GTA and CSA changes and risk threshold of RCTs,as well as the protective factors and risk factors.Methods:In this study,130 individuals from May 2019 to December 2020 were recruited.According to Southern California Orthopedic Institute(SCOI)classifica-tion,the individuals were divided into four groups retrospectively:Group A,negative control group;Group B,partial tears(articular side);Group C,partial tears(bursal side);Group D,full-thickness tears.GTA and CSA were measured respectively on true anteroposterior position X-ray of shoulder with arm in neutral rotation and performed by the same trained technician team in single-blind.The correla-tions between RCTs and relevant factors were analyzed.Results:According to the area under the receiver operating characteristic curve(AUC),GTA and CSA of RCTs(Groups B,C and D)were 0.736 and 0.673 with 95％confidence interval(CI),the cut-off value of GTA and CSA of RCTs were 70.5° and 39.5° respectively.Comparing with the control group,RCTs groups had significant statistical differences in age and body mass index(BMI)(P＜0.05),especially the full-thickness RCTs(Group D),which was older than Groups A,B and C(P＜0.05,cut-off value:56.5 years old)and shorter than Groups A and B(P＜0.05,cut-off value:1.58 m).Analyzed from scatter plot and regression analysis,there was no linear correlation between GTA and CSA.There were no significant differences in gender,dominant shoulders and smoking between the RCTs groups and the control group(P＞0.05).Conclusion:Larger GTA(＞70.5°)and CSA(＞39.5°)would be highly predictive in diagnosing RCTs without linear cor-relation,and GTA has a higher diagnostic value in contrast.Subacromial impingement and shoulder de-generation occurred before RCTs.Patients with age＞56.5 years and height＜1.58 m were more likely to develop disease of full-thickness RCTs and no statistic differences in weight and BMI.Gender,domi-nant shoulder and smoking were neither risk factors nor protective factors.

Objective Through molecular genetics analysis of a calculi family lineage,this study aims to explore its pathogenesis and the association between genotypes and phenotypes.Methods A retrospective analysis was conducted on a calculi family lineage admitted to Tongji Hospital,affiliated with Tongji Medical College,Huazhong University of Science and Technology.Clinical data and peripheral blood samples of the affected children and some family members were collected.Whole exome sequencing was performed,followed by Sanger sequencing to validate the candidate variants.Results Among the 38 family members across four generations,10 members were diagnosed with calculi disease.The second generation,member 2(Ⅱ-2),third generation,member 2(Ⅲ-2),and third generation,member 4(Ⅲ-4)suffered from recurrent multiple kidney stones and gallstones.The second generation,member 6(Ⅱ-6),second generation,member 13(Ⅱ-13),and third generation,member 5(Ⅲ-5)had recurrent multiple kidney stones alone,while first generation,member 2(Ⅰ-2),second generation,member 4(Ⅱ-4),second generation,member 8(Ⅱ-8),and second generation,member 11(Ⅱ-11)only had gallstones.No other family members exhibited any signs of kidney or gallbladder involvement.Ⅱ-2 was diagnosed in 2018 with end-stage renal disease stage 5,grade 3 hypertension and gallstones,urinary amino acid high-performance liquid chromatography analysis indicated elevated urinary cystine.This member had a history of recurrent multiple kidney stones and recurrent urinary tract infections for over 30 years,with multiple histories of ureteroscopic stone removal.Genetic analysis revealed that Ⅱ-2,Ⅲ-2,Ⅲ-4,Ⅲ-5 and Ⅳ-1 all carry a heterozygous mutation in exon 10 of the solute carrier family 3 member 1(SLC3A1)gene,c.1889G＞A(p.Gly630Asp).The third generation,member 1(Ⅲ-1),and fourth generation,member 2(Ⅳ-2),are wild type.This mutation shows a phenomenon of family co-segregation.Conclusions The heterozygous mutation of SLC3A1 gene,c.1889G＞A,may be the genetic cause of calculi disease in multiple members of this family lineage.Recurrent multiple kidney stones and/or gallstones require high attention to genetic etiology.It is recommended to perform genetic analysis on calculi family lineages and patients with early-onset calculi disease.