Objective:To explore the clinical and genetic characteristics of fetuses with Kabuki syndrome (KS) and their genotype-phenotype correlation.Methods:A retrospective analysis was carried out on the prenatal manifestations and results of genetic testing of six KS fetuses diagnosed by whole-exome sequencing (WES). The findings were compared with 28 prenatally diagnosed KS cases reported in the literature to summarize the prenatal features of KS. This study has been approved by the Ethics Committee of Maternal and Child Health Care Hospital of Hubei Province (Ethics No.: 2025-141-01).Results:Prenatal ultrasound findings in KS fetuses showed high heterogeneity. The most common abnormalities were cardiac (23/35, 65.7%) and renal (20/35, 57.1%), which are often accompanied by amniotic fluid abnormalities (5/35, 14.3%), single umbilical artery (5/35, 14.3%), and fetal hydrops (4/35, 11.4%). Among the six fetuses from our center, all were identified by WES to harbor pathogenic variants of the KMT2D gene, and all of which were de novo. These included 3 frameshift variants, 2 nonsense variant, and 1 missense variant, among which 4 were unreported previously. Conclusion:This study has expanded the mutational spectrum of the KMT2D gene. Prenatal ultrasound findings of KS lack specificity, though multi-system anomalies or specific soft markers may indicate KS. WES is an effective tool for the diagnosis, and KS should be included in the differential diagnosis list for prenatal cardiac and renal abnormalities.

OBJECTIVE: To explore the clinical and genetic characteristics of fetuses with Kabuki syndrome (KS) and their genotype-phenotype correlation. METHODS: A retrospective analysis was carried out on the prenatal manifestations and results of genetic testing of six KS fetuses diagnosed by whole-exome sequencing (WES). The findings were compared with 28 prenatally diagnosed KS cases reported in the literature to summarize the prenatal features of KS. This study has been approved by the Ethics Committee of Maternal and Child Health Care Hospital of Hubei Province (Ethics No.: 2025-141-01). RESULTS: Prenatal ultrasound findings in KS fetuses showed high heterogeneity. The most common abnormalities were cardiac (23/35, 65.7%) and renal (20/35, 57.1%), which are often accompanied by amniotic fluid abnormalities (5/35, 14.3%), single umbilical artery (5/35, 14.3%), and fetal hydrops (4/35, 11.4%). Among the six fetuses from our center, all were identified by WES to harbor pathogenic variants of the KMT2D gene, and all of which were de novo. These included 3 frameshift variants, 2 nonsense variant, and 1 missense variant, among which 4 were unreported previously. CONCLUSION This study has expanded the mutational spectrum of the KMT2D gene. Prenatal ultrasound findings of KS lack specificity, though multi-system anomalies or specific soft markers may indicate KS. WES is an effective tool for the diagnosis, and KS should be included in the differential diagnosis list for prenatal cardiac and renal abnormalities.
Humans ; Hematologic Diseases/diagnostic imaging* ; Face/diagnostic imaging* ; Female ; Vestibular Diseases/diagnostic imaging* ; Abnormalities, Multiple/diagnostic imaging* ; Pregnancy ; Ultrasonography, Prenatal ; Adult ; Neoplasm Proteins/genetics* ; Retrospective Studies ; DNA-Binding Proteins/genetics* ; Male ; Exome Sequencing ; Fetus/diagnostic imaging* ; Genetic Association Studies ; Mutation

Objective:To explore the clinical and genetic characteristics of fetuses with Kabuki syndrome (KS) and their genotype-phenotype correlation.Methods:A retrospective analysis was carried out on the prenatal manifestations and results of genetic testing of six KS fetuses diagnosed by whole-exome sequencing (WES). The findings were compared with 28 prenatally diagnosed KS cases reported in the literature to summarize the prenatal features of KS. This study has been approved by the Ethics Committee of Maternal and Child Health Care Hospital of Hubei Province (Ethics No.: 2025-141-01).Results:Prenatal ultrasound findings in KS fetuses showed high heterogeneity. The most common abnormalities were cardiac (23/35, 65.7%) and renal (20/35, 57.1%), which are often accompanied by amniotic fluid abnormalities (5/35, 14.3%), single umbilical artery (5/35, 14.3%), and fetal hydrops (4/35, 11.4%). Among the six fetuses from our center, all were identified by WES to harbor pathogenic variants of the KMT2D gene, and all of which were de novo. These included 3 frameshift variants, 2 nonsense variant, and 1 missense variant, among which 4 were unreported previously. Conclusion:This study has expanded the mutational spectrum of the KMT2D gene. Prenatal ultrasound findings of KS lack specificity, though multi-system anomalies or specific soft markers may indicate KS. WES is an effective tool for the diagnosis, and KS should be included in the differential diagnosis list for prenatal cardiac and renal abnormalities.

Objective:To study the genetic variation and distribution characteristics of thalassemia in people of childbearing age in Hubei Province, and to provide clinical basis for the local government decision-making departments to formulate and promote appropriate policies for prevention and control of thalassemia.Methods:Venous blood samples were collected from 44 849 people of childbearing age in hospitals in Hubei Province from May 13, 2019 to August 17, 2021. PCR-flow fluorescence hybridization and PCR+diversion hybridization were used to screen thalassemia genes. Spouses of those who tested positive were also tested for thalassemia genes. When both spouses carried the same type of thalassemia gene, the amniotic fluid of pregnant women was extracted for prenatal diagnosis and followed up.Results:Among the 44 849 people of childbearing age, 2 286 cases were diagnosed as thalassemia gene carriers through genetic testing, and the total detection rate was 5.10% (2 286/44 849). Among them, 1 488 cases were diagnosed as α-thalassemia, and the detection rate was 3.32% (1 488/44 849); 767 cases were diagnosed as β-thalassemia, and the detection rate was 1.71% (767/44 849); 31 cases were diagnosed as α-thalassemia combined with β-thalassemia, and the detection rate was 0.07% (31/44 849). The top three genotypes of α-thalassemia were -α 3.7/αα, -- SEA/αα, and -α 4.2/αα, accounting for 58.06% (864/1 488), 26.14% (389/1 488), and8.74% (130/1 488), respectively. The top three genotypes of β-thalassemia were β IVS-Ⅱ-654/β N, β CD41-42/β N, and β CD17/β N, accounting for 41.72% (320/767), 21.25% (163/767), and 16.04% (123/767), respectively. The main genotypes of α-thalassemia combined with β-thalassemia were -α 3.7/αα complex β IVS-Ⅱ-654/β N and -α 3.7/αα complex β CD41-42/β N, accounting for 29.03% (9/31) and 16.13% (5/31), respectively. A total of 59 people of childbearing age were conducted prenatal diagnosis, among fetus, there were 4 cases of severe thalassemia (2 cases of severe α-thalassemia, 2 cases of severe β-thalassemia), 5 cases of intermediate α-thalassemia, 5 cases of intermediate or severe β-thalassemia, 19 cases of mild thalassemia (8 cases of mild α-thalassemia, 11 cases of mild β-thalassemia), 13 cases of stationary α-thalassemia, and 1 case of stationary α-thalassemia combined with mild β-thalassemia, there were 12 cases without α-thalassemia or β-thalassemia genes. After follow-up, 4 cases of severe thalassemia, 2 cases of intermediate α-thalassemia, and 5 cases of intermediate or severe β-thalassemia were terminated pregnancy by the joint decision of both parents. Conclusions:In Hubei Province, the detection rate of thalassemia is high, and α-thalassemia is the main mutation type. The combination of thalassemia gene screening and prenatal diagnosis is of great significance in reducing the birth rate of children with thalassemia.

OBJECTIVE：To establish a method for simultaneous determination of gallic acid ，protocatechuic acid ，mangiferin， homomangiferin，hyperoside and isoquercetin in Mangguo zhike tablets. METHODS ：HPLC was adopted. The determination was performed on Phenomenex Gemini C 18 column with mobile phase consisted of 0.1％ phosphoric acid water solution-acetonitrile （gradient elution ）at the flow rate of 1.0 mL/min. The detection wavelength was set at 258 nm. The column temperature was set at 30 ℃，and sample size was 5 μL. Gallic acid was used as the internal substance，and the correction factors of gallic acid ， protocatechuic acid ，mangiferin，homomangiferin，hyperoside and isoquercetin were calculated ；the results of quantitative analysis of multi-components by single marker method （QAMS） were compared with those of external standard method （ESM）. RESULTS：The linear range of gallic acid ，protocatechuic acid ，mangiferin，homomangiferin，hyperoside and isoquercetin were 45.75-183 0 μg/mL（r＝0.999 9），2.525-101.0 μg/mL（r＝0.999 9），65.33-261 3 μg/mL（r＝0.999 6），9.058-362.3 μg/mL（r＝ 0.999 9），3.885-155.4 μg/mL（r＝0.999 9），1.870-74.8 μg/mL（r＝0.999 9），respectively. The limits of quantification were 0.571， 0.643，1.053，0.854，0.830，1.500 μg/mL，respectively. The limits of detection were 0.171，0.193，0.316，0.256，0.249，0.450 μg/mL， respectively. RSDs of precision ，stability and reproducibility tests were all lower than 3％ . The average recoveries were 98.8％-101.9％（RSD＝1.3％ ，n＝6），94.3％-101.5％（RSD＝3.3％，n＝6），97.9％-100.5％（RSD＝0.9％，n＝6），98.2％- 101.6％ （RSD＝1.2％，n＝6），102.3％-106.1％（RSD＝1.3％，n＝6）， 96.6％ -99.3％（RSD＝1.0％ ，n＝6），respectively. RCFs of 73） protocate-chuic acid ， mangiferin， homomangiferin，hyperoside and isoquercetin were 0.568 3，0.500 3，0.687 6， 0.939 1 and 0.826 3，using gallic acid as internal substance . The content ranges of protocatechuic acid and other 4 com- ponents measured by QAMS were 0.197-0.440，3.262-11.250， 0.201-1.196，0.168-0.381，0.115-0.293 mg/tablet，respectively. The content ranges measured by ESM were 0.198-0.441，3.239-11.570，0.206-1.194，0.171-0.380，0.119-0.298 mg/tablet， respectively. By comparing the content determination results by QAMS and ESM ，the relative errors were －3.80％-0.74％，and there was no statistical significance （P＞0.05）. CONCLUSIONS ：The established QAMS method is accurate ，reliable and repeatable，and can be used for content determination of 6 medicinal components in Mangguo zhike tablets.

OBJECTIVE：To study the spectrum-effect relationship of HPLC finger print of different polar parts of Ampelopsis grossedentata with its in vivo anti-inflammatory effect. METHODS ：A. grossedentata was reflux extracted with 70％ ethanol，then extracted with petroleum ether ，chloroform，ethyl acetate and water saturated n-butanol；or it was directly decocted with water and then concentrated to obtain different polar parts. The xylene-induced mice ear swelling model was established ；using dexamethasone as positive control ，anti-inflammatory activity of different polar parts of A. grossedentata was investigated. Fingerprints of different polar parts of A. grossedentata were established by HPLC. The determination was performed on Poroshell 120 EC-C18 column with mobile phase consisted of acetonitrile- 0.1％ phosphoric acid solution （gradient elution ）at the flow rate of 1 mL/min. The column temperature was 25 ℃. The detection wavelength was set at 365 nm，and sample size was 5 μL. The grey ralational analysis method was used to analyze the spectrum-effect relationship of HPLC fingerprint common peaks of different polar parts of A. grossedentata with its anti-inflammatory effect. The correlation coefficient and correlation degree were calculated and ranked. RESULTS：Anti-inflammatory experiment showed that the anti-inflammatory effects of 70％ ethanol extraction part ，ethyl acetate extraction part and water extraction part were the most significant （inhibitory rates of ear swelling were 54.07％，30.54％， 30.45％）. Five common peaks were determined in HPLC fingerprints of different polar parts from A. grossedentata . The spectrum-effect analysis results showed that the correlation of5 common peaks were higher than 0.6；among them ，peak 3 and peak 2 （dihydromyricetin） had the strongest anti- inflammatory effect ，and their correlation degrees were both mail：123745789@qq.com greater than 0.8. CONCLUSIONS ： The anti-inflammatory effect of A. grossedentata on xylene-induced ear swelling in mice is the result of multi-comp onent synergy ； unknown substance of peak 3 and dihydromyricetin may be the main active components of A. grossedentata .

OBJECTIVE:To establish a method for the simultaneous determination of syringinand and pedunculoside in Zhuang medicine Yuyang powder. METHODS:HPLC was performed on the column of Agilent ODS with mobile phase of acetonitrile-water (gradient elution)at a flow rate of 1 ml/min,the detection wavelength was 210 nm,the column temperature was 30℃,and the in-jection volume was 5μl. RESULTS:The linear range was 0.71-3.55μg for syringin(r=0.999 7)and 1.62-8.10μg for pedunculoside (r=0.999 8), respectively;RSDs of precision, stability and reproducibility tests were lower than 3%;recoveries were 100.8%-104.9%(RSD=1.7%,n=6) and 96.0%-100.8%(RSD=2.2%,n=6). CONCLUSIONS:The method is simple,stable and reproducible,and can be used for the simultaneous determination of syringinand and pedunculoside in Zhuang medicine Yuyang powder.

Objective To investigate the preventive and therapeutic effects of vesiculation moxibustion on rat hepatic fibrosis (HF).Methods Thirty rats were randomly allocated to treatment, model and control groups, 10 rats each. A rat model of HF was made by intraperitoneal injection of carbon tetrachloride. At three weeks after model making, the treatment group was treated by vesiculation moxibustion 6 hrs twice a week, for a total of 4 weeks. Body weight, liver wet weight, liver index, serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) contents, and serum and liver typeⅢ procollagen (PCⅢ), typeⅣ collagen (Ⅳ-C), hyaluronic acid (HA) and laminin (LN) contents were measured in every group of rats.Results There were statistically significant differences in HF severity grade and HF tissue collagen analysis indices between the model group of rats and the treatment group or the control group (P<0.01,P<0.05). There were statistically significant differences in body weight, liver wet weight and liver index between the model and control groups of rats (P<0.01). There were statistically significant differences in liver wet weight and liver index between the treatment and model groups of rats (P<0.01). There were statistically significant differences in serum ALT and AST contents, and serum and liver PCⅢ,Ⅳ-C, HA and LN contents between the model and control groups of rats (P<0.01,P<0.05). There were statistically significant differences in serum ALT and AST contents, and serum and liverⅣ-C, HA and LN contents between the treatment and model groups of rats (P<0.01,P<0.05). There was a statistically significant difference in serum PCⅢ content between the treatment and model groups of rats (P<0.01).Conclusion Vesiculation moxibustion has some preventive and therapeutic effects on rat hepatic fibrosis.

Physiology experiment can effectively stimulate the enthusiasm of the students to explore the scientific truth by providing the opportunity to observe real physiological phenomena,analyze,discuss and solve practical problems.Lab-based learning (LBL) and theory teaching have complementary advantages,promoting each other and can maximize the use of teaching resources.During the process of physiological LBL,training of students' thinking ability by creating problem situation,inspiration and guidance,carrying out exploring experiments can develop students' intelligence,promote innovation,improve the teaching effect.

BACKGROUND:Bone marrow mesenchymal stem cels have a therapeutic effect on acute lung injury, but the mechanism is unclear. If the mechanism is understood, the majority of patients with acute lung injury can obtain a benefit. OBJECTIVE:To explore the possible mechanism underlying bone marrow mesenchymal stem cels in the treatment of acute lung injury with sepsis in rats. METHODS: (1) Thirty-six adult Wistar rats were randomly divided into three groups, sham operation group (sham group), sepsis group and bone marrow mesenchymal stem cels group (cel treatment group). In the sepsis and cel treatment groups, animal models of sepsis with acute lung injury were established by cecal ligation and puncture, while in the sham group, the cecum was not ligated and punctured. Then, 1 mL normal saline was injected via the femoral vein in the sepsis and sham groups, and 1 mL bone marrow mesenchymal stem cel suspension (1×109/L) was injected into the cel treatment group. After 6 hours, interleukin 10 and macrophage inflammatory protein-2 levels in serum were measured in the three groups. Lung tissues were taken for pathological observation using hematoxylin-eosin staining. (2) Rat alveolar macrophages were obtained by bronchoalveolar lavage, seeded into 24-wel culture plates, and divided into three groups: control group (group A), sepsis model group (group B) and intervention group of bone marrow mesenchymal stem cels (group C). Normal saline, septic plasma, and co-intervention of septic plasma and mesenchymal stem cels were used in the groups A, B, C, respectively. Then, cels in the three groups were cultured in a 5% CO2 incubator at 37℃ for 1 hour. After that, alveolar macrophages were taken to detect whether nuclear factor-κB (P65) protein entered into the nucleus using laser scanning confocal microscopy. RESULTS AND CONCLUSION: (1) The results of animal experiments showed that compared with the sham group, the macrophage inflammatory protein-2 levels in the sepsis group and cel treatment group were significantly increased (P < 0.05), but the macrophage inflammatory protein-2 level in the cel treatment group was significantly lower than that in the sepsis group (P < 0.05); there were no significant differences in serum interleukin 10 levels among the three groups (P > 0.05); inflammatory cel infiltration, interstitial pulmonary edema and pulmonary hemorrhage existed in the sepsis and cel treatment groups, but these symptoms were significantly reduced in the cel treatment group compared with the sepsis group. (2) Results from cel experiments showed that compared with the group A, in group B and group C, the number of nuclear factor-κB (P65) proteins into the nucleus was significantly higher (P < 0.05), but it was lower in the group C than the group B (P < 0.05). These findings indicate that bone marrow mesenchymal stem cels in acute lung injury with sepsis can regulate nuclear factor-κB (P65) protein of alveolar macrophages into the nucleus, reduce expression of macrophage inflammatory protein-2, and thereby play a protective role in the lungvia reducing neutrophil infiltration. Temporarily, this study cannot explain whether bone marrow mesenchymal stem cels have an effect on interleukin 10.