Objective To establish quality evaluation method of Andrographis paniculata standard decoction by UPLC. Methods 21 batches of Andrographis paniculata standard decoctions were prepared according to the standardization method of TCM decoction pieces. The UPLC characteristic chromatograms analysis method was established. With andrographolide as a reference, quantitative analysis of multi-components by single marker （QAMS） was established for new neoandrographolide, 14 deoxyandrographolide and dehydrated andrographolide. The results were compared with the external standard method （ESM） to determine the accuracy of the method. Results Similarity Evaluation System for Chromatographic Fingerprint of TCM （2012 edition） was used to analyze and compare the characteristic chromatograms, and seven common peaks were determined and five were identified including luteolin-7-O-β-D-glucuronide, andrographolide, neoandrographolide, 14-deoxyandrographolide and dehydroandrographolide. The RSDs of content results of each component by QAMS and ESM were all within 3%. Conclusion The determination method was reliable and accurate, which could be used to reflect the intrinsic quality of Andrographis paniculata standard decoction comprehensively and provide the basis for the quality evaluation of Andrographis paniculata formula granules and other preparations.

Objective:To explore the effect and mechanism of Chlorfortunone A(ChlA) in the treatment of diabetic nephropathy(DN) in mice.Methods:The DN model mice were assigned to DN, low-dose ChlA(ChlAL) and high-dose ChlA(ChlAH), and the normal control groups(Ctrl). Kidney tissue was analyzed via HE and Masson staining, and urine albumin, fasting blood glucose and kidney weight were measured. Collagen1 and α-SMA proteins were detected in renal tissues. The level of GSH-px, SOD, CAT, and TGF-β were detected. The TGF-β/Smad2 pathway in kidney tissue was detected. The mechanism was verified by setting the high glucose+ ChlA+ TGF-β group in MPC-5 cells. The proliferation of the cells and DCFDA staining were detected.Results:Compared to the Ctrl group, the DN group had significantly higher UACR and kidney weight( P<0.001). High-dose ChlA reduced UACR and kidney weight( P<0.05), with no effect on blood glucose( P>0.05). Masson staining showed reduced fibrosis with ChlA treatment. Collagen I and α-SMA expressions were significantly higher in DN( P<0.001) and decreased with ChlA treatment( P<0.05). GSH-px, SOD, and CAT levels were lower in DN( P<0.001), while TGF-β was elevated( P<0.001); ChlA increased antioxidant enzymes and decreased TGF-β( P<0.05). The TGF-β/Smad2 pathway was upregulated in DN( P<0.001) and inhibited by ChlA( P<0.001). In vitro, ChlA reduced cell proliferation( P<0.05) and increased ROS levels( P<0.001). Conclusions:ChlA alleviates kidney injury and fibrosis in DN mice, reduces oxidative stress, which may be related to the inhibition of the TGF-β/Smad2 pathway.

Objective:To observe the clinical effect of comprehensive reconstruction of grades Ⅰ-Ⅱ thumb and finger defects with hallux osteo-onychocutaneous flaps of great toe.Methods:This is a retrospective study. From June 2020 to December 2023, comprehensive reconstruction surgery for Grade Ⅰ-Ⅱ digital defect were performed with hallux osteo-onychocutaneous flaps of great toe for 3 thumbs and 7 fingers in 9 patients in the Department of Hand and Microsurgery of Baoji Third Hospital. Causes of digital injury were: 4 of machine crush, 3 of electric saw cut, 1 of door crush, and 1 of electrical burn. There were 6 grade I digital defects (beyond the nail root) and 4 grade Ⅱ defects (last segment of digit). The defects of the digits were reconstructed by taking references of the shape and structure of the contralateral normal thumbs and fingers. Then the hallux osteo-onychocutaneous flaps of great toe were designed and harvested accordingly from the left and right great toes. Donor sites were covered by skin grafting or local dressing change. One patient was treated in emergency surgery, 6 in sub-emergency surgery and 2 in elective surgery. Integrated perioperative patient management was provided to all of the patients. Postoperative follow-ups were conducted through the visit of outpatient clinic, telephone calls or WeChat interviews. Flap survival, appearance and sensation recovery were evaluated according to the Evaluation Standard of Upper Limb Functional of Hand Surgery of Chinese Medical Association.Results:Vascular insufficiency of 1 digit occurred in surgery, and relieved by local treatment with lidocaine and warm saline. All 10 digits successfully survived, and all donor sites healed spontaneously. The postoperative follow-up period was 10 to 30 months, with an average of 18 months. One transferred nail was found in poor appearance (not flat), the rest of the reconstructed digits were good in appearance and function. The nail, finger print and fine sensation (TPD 5~8 mm), as well as digital flexion, extension, grasping and opposition of the reconstructed digits were all good. According to the Evaluation Standard of Upper Limb Functional of Hand Surgery of Chinese Medical Association, at the last follow-up visit, 5 digits were in excellent, 4 in good and 1 in fair.Conclusion:The comprehensive reconstruction of grades Ⅰ-Ⅱ digital defects with hallux osteo-onychocutaneous flap of great toe is an ideal surgical technique that can reconstruct the nail, finger print and sensation of a digit, with good postoperative function as well as an aesthetic and realistic appearance.

OBJECTIVE: To explore the clinical characteristics and genetic etiology of a child with Neurofibromatosis-Noonan syndrome (NFNS). METHODS: A child with NFNS who was treated at the Department of Endocrinology of Hangzhou Children's Hospital in January 2024 was selected as the study subject. Clinical data of the child was collected by retrospective analysis. Peripheral venous blood samples (2 mL each) were collected from the child and his parents. Genomic DNA was extracted, and trio-whole exome sequencing (Trio-WES) of the family was carried out. Sanger sequencing was used to perform family verification on the candidate variants. The identified variants were classified for pathogenicity according to the Standards and Guidelines for the Interpretation of Sequence Variants established by the American College of Medical Genetics and Genomics (ACMG) (hereafter referred to as the "ACMG guidelines"). This study has been approved by the Medical Ethics Committee of Hangzhou Children's Hospital (Ethics No. 2021-06). RESULTS: The child was a 7-year and 4-month-old male. He has short stature, numerous café-au-lait spots on the neck and trunk, and special facial features such as a full forehead, wide interpupillary distance, a low nasal bridge, and low-set ears. The results of Trio-WES showed that the he had harbored the NF1 gene c.3773G>T (p.W1258L) mutation, which was verified by Sanger sequencing to be de novo in origin. The NF1 gene was associated with NFNS, which has an autosomal dominant inheritance. According to the ACMG guidelines, this variant was judged to be a likely pathogenic variant (PS2+PM2+PP3+PP2). No pathogenic variant in genes associated with Noonan syndrome, such as PTPN11, SOS1, RAF1, RIT1, and KRAS, was found. CONCLUSION The child with NFNS has clinical features such as short stature, special facial features, and café-au-lait spots. The c.3773G>T (p.W1258L) variation in the NF1 gene may be the genetic etiology of the NFNS child in this study. The results of this study has enriched the variation spectrum of the NF1 gene.
Child ; Humans ; Male ; Exome Sequencing ; Mutation ; Neurofibromatosis 1/genetics* ; Neurofibromin 1/genetics* ; Noonan Syndrome/genetics*

Objective:To explore the clinical characteristics and genetic etiology of a child with Neurofibromatosis-Noonan syndrome (NFNS).Methods:A child with NFNS who was treated at the Department of Endocrinology of Hangzhou Children′s Hospital in January 2024 was selected as the study subject. Clinical data of the child was collected by retrospective analysis method. Peripheral venous blood samples (2 mL each) were collected from the child and his parents. Genomic DNA was extracted, and trio whole exome sequencing (Trio-WES) of the family was carried out. Sanger sequencing was used to perform family verification on the candidate variants. The identified variants were classified for pathogenicity according to the Standards and Guidelines for the Interpretation of Sequence Variants established by the American College of Medical Genetics and Genomics (ACMG) (hereafter referred to as the " ACMG guidelines" ). This study has been approved by the Medical Ethics Committee of Hangzhou Children′s Hospital (Ethics No. 2021-06).Results:The child was a 7-year and 4-month-old male. He has short stature, numerous café-au-lait spots on the neck and trunk, and special facial features such as a full forehead, wide interpupillary distance, a low nasal bridge, and low-set ears.The results of Trio-WES showed that he has harbored a NF1 gene c. 3773G>T (p.W1258L) mutation, which was verified by Sanger sequencing to be de novo in origin. The NF1 gene child was associated with NFNS, which was an autosomal dominant inheritance. According to the ACMG guidelines, this variant was judged to be a likely pathogenic variant (PS2+ PM2+ PP3+ PP2). No pathogenic variant in genes associated with Noonan syndrome, such as those in PTPN11, SOS1, RAF1, RIT1, and KRAS, was found. Conclusion:The child with NFNS has clinical features such as short stature, special facial features, and café-au-lait spots. The c. 3773G>T (p.W1258L) variation in the NF1 gene may be the genetic etiology of the NFNS child in this study. The results of this study has enriched the variation spectrum of the NF1 gene.

Objective:To explore the clinical characteristics and genetic etiology of a child with Neurofibromatosis-Noonan syndrome (NFNS).Methods:A child with NFNS who was treated at the Department of Endocrinology of Hangzhou Children′s Hospital in January 2024 was selected as the study subject. Clinical data of the child was collected by retrospective analysis method. Peripheral venous blood samples (2 mL each) were collected from the child and his parents. Genomic DNA was extracted, and trio whole exome sequencing (Trio-WES) of the family was carried out. Sanger sequencing was used to perform family verification on the candidate variants. The identified variants were classified for pathogenicity according to the Standards and Guidelines for the Interpretation of Sequence Variants established by the American College of Medical Genetics and Genomics (ACMG) (hereafter referred to as the " ACMG guidelines" ). This study has been approved by the Medical Ethics Committee of Hangzhou Children′s Hospital (Ethics No. 2021-06).Results:The child was a 7-year and 4-month-old male. He has short stature, numerous café-au-lait spots on the neck and trunk, and special facial features such as a full forehead, wide interpupillary distance, a low nasal bridge, and low-set ears.The results of Trio-WES showed that he has harbored a NF1 gene c. 3773G>T (p.W1258L) mutation, which was verified by Sanger sequencing to be de novo in origin. The NF1 gene child was associated with NFNS, which was an autosomal dominant inheritance. According to the ACMG guidelines, this variant was judged to be a likely pathogenic variant (PS2+ PM2+ PP3+ PP2). No pathogenic variant in genes associated with Noonan syndrome, such as those in PTPN11, SOS1, RAF1, RIT1, and KRAS, was found. Conclusion:The child with NFNS has clinical features such as short stature, special facial features, and café-au-lait spots. The c. 3773G>T (p.W1258L) variation in the NF1 gene may be the genetic etiology of the NFNS child in this study. The results of this study has enriched the variation spectrum of the NF1 gene.

Objective:To explore the effect and mechanism of Chlorfortunone A(ChlA) in the treatment of diabetic nephropathy(DN) in mice.Methods:The DN model mice were assigned to DN, low-dose ChlA(ChlAL) and high-dose ChlA(ChlAH), and the normal control groups(Ctrl). Kidney tissue was analyzed via HE and Masson staining, and urine albumin, fasting blood glucose and kidney weight were measured. Collagen1 and α-SMA proteins were detected in renal tissues. The level of GSH-px, SOD, CAT, and TGF-β were detected. The TGF-β/Smad2 pathway in kidney tissue was detected. The mechanism was verified by setting the high glucose+ ChlA+ TGF-β group in MPC-5 cells. The proliferation of the cells and DCFDA staining were detected.Results:Compared to the Ctrl group, the DN group had significantly higher UACR and kidney weight( P<0.001). High-dose ChlA reduced UACR and kidney weight( P<0.05), with no effect on blood glucose( P>0.05). Masson staining showed reduced fibrosis with ChlA treatment. Collagen I and α-SMA expressions were significantly higher in DN( P<0.001) and decreased with ChlA treatment( P<0.05). GSH-px, SOD, and CAT levels were lower in DN( P<0.001), while TGF-β was elevated( P<0.001); ChlA increased antioxidant enzymes and decreased TGF-β( P<0.05). The TGF-β/Smad2 pathway was upregulated in DN( P<0.001) and inhibited by ChlA( P<0.001). In vitro, ChlA reduced cell proliferation( P<0.05) and increased ROS levels( P<0.001). Conclusions:ChlA alleviates kidney injury and fibrosis in DN mice, reduces oxidative stress, which may be related to the inhibition of the TGF-β/Smad2 pathway.

Objective:To observe the clinical effect of comprehensive reconstruction of grades Ⅰ-Ⅱ thumb and finger defects with hallux osteo-onychocutaneous flaps of great toe.Methods:This is a retrospective study. From June 2020 to December 2023, comprehensive reconstruction surgery for Grade Ⅰ-Ⅱ digital defect were performed with hallux osteo-onychocutaneous flaps of great toe for 3 thumbs and 7 fingers in 9 patients in the Department of Hand and Microsurgery of Baoji Third Hospital. Causes of digital injury were: 4 of machine crush, 3 of electric saw cut, 1 of door crush, and 1 of electrical burn. There were 6 grade I digital defects (beyond the nail root) and 4 grade Ⅱ defects (last segment of digit). The defects of the digits were reconstructed by taking references of the shape and structure of the contralateral normal thumbs and fingers. Then the hallux osteo-onychocutaneous flaps of great toe were designed and harvested accordingly from the left and right great toes. Donor sites were covered by skin grafting or local dressing change. One patient was treated in emergency surgery, 6 in sub-emergency surgery and 2 in elective surgery. Integrated perioperative patient management was provided to all of the patients. Postoperative follow-ups were conducted through the visit of outpatient clinic, telephone calls or WeChat interviews. Flap survival, appearance and sensation recovery were evaluated according to the Evaluation Standard of Upper Limb Functional of Hand Surgery of Chinese Medical Association.Results:Vascular insufficiency of 1 digit occurred in surgery, and relieved by local treatment with lidocaine and warm saline. All 10 digits successfully survived, and all donor sites healed spontaneously. The postoperative follow-up period was 10 to 30 months, with an average of 18 months. One transferred nail was found in poor appearance (not flat), the rest of the reconstructed digits were good in appearance and function. The nail, finger print and fine sensation (TPD 5~8 mm), as well as digital flexion, extension, grasping and opposition of the reconstructed digits were all good. According to the Evaluation Standard of Upper Limb Functional of Hand Surgery of Chinese Medical Association, at the last follow-up visit, 5 digits were in excellent, 4 in good and 1 in fair.Conclusion:The comprehensive reconstruction of grades Ⅰ-Ⅱ digital defects with hallux osteo-onychocutaneous flap of great toe is an ideal surgical technique that can reconstruct the nail, finger print and sensation of a digit, with good postoperative function as well as an aesthetic and realistic appearance.

Objective:To study the clinical value of serum microRNA-34a (miR-34a) and aryl hydrocarbon receptor nuclear transcript-like protein 1 (BMAL1) in cervical cancer and their relationship with high-risk human papillomavirus (HR-HPV) infection.Methods:The clinical data of 76 patients with cervical cancer and 50 patients with benign cervical diseases in the Affiliated Hospital of Jining Medical University from January to December 2022 were retrospectively analyzed. The expression levels of serum miR-34a and BMAL1 were detected by real-time fluorescence quantitative polymerase chain reaction, and HR-HPV infection was detected by flow fluorescent hybridization. The patients were followed up until December 2023, and the death and poor prognosis (death, tumor recurrence and progression and severe complications at 1-year of follow-up) were recorded. The receiver operating characteristic (ROC) curve was used to analyze the efficacy of miR-34a, BMAL1 and related indexes in evaluating the poor prognosis in patients with cervical cancer at 1-year. Multivariate Cox regression analysis was used to analyze the independent risk factors for death in patients with cervical cancer. The Kaplan-Meier survival curve was used to analyze the relationship between miR-34a, BMAL1 expression and survival period, and the log-rank test was used for comparison.Results:The expression level of serum miR-34a in patients with cervical cancer was significantly lower than that in patients with benign cervical lesions (0.46 ± 0.08 vs. 0.67 ± 0.11), the expression level of serum BMAL1 was significantly higher than that in patients with benign cervical lesions (0.58 ± 0.07 vs. 0.41 ± 0.07), and there were statistical differences ( t= 12.40 and 13.34, P<0.01). The expression levels of serum miR-34a and BMAL1 in patients with cervical cancer were associated with tumor differentiation, myometrial invasion depth, lymph node metastasis, distant metastasis and International Federation of Gynecology and Obstetrics (FIGO) stage, and there were statistical differences ( P<0.05 or<0.01); they were not associated with age, menopause and pathological type, and there were no statistical differences ( P>0.05). In patients with cervical cancer, the expression level of miR-34a in patients with HR-HPV positive infection (60 cases) was significantly lower than that in patients with HR-HPV negative infection (16 cases): 0.41 ± 0.07 vs. 0.49 ± 0.08, the expression level of BMAL1 was significantly higher than that in patients with HR-HPV negative infection: 0.65 ± 0.09 vs. 0.53 ± 0.06, and there were statistical differences ( t = 3.68 and 4.24, P<0.05 or<0.01). In patients with benign cervical diseases, there were no statistical differences in the expression levels of miR-34a and BMAL1 between patients with HR-HPV positive infection (7 cases) and patients with HR-HPV negative infection (43 cases) ( P>0.05). ROC curve analysis result showed that miR-34a combined with BMAL1 had the highest sensitivity (90.4%), specificity (89.9%) and area under curve (0.911) in assessing the 1-year poor prognosis in patients with cervical cancer ( P<0.01), and the optimal cutoff values of miR-34a and BMAL1 expression level were ≤0.39 and ≥0.64. Multivariate Cox regression analysis result showed that poor differentiation, myometrial invasion depth ≥1/2, lymph node metastasis, distant metastasis, FIGO stage Ⅲ+Ⅳ, miR-34a expression level ≤0.46 and BMAL1 expression level ≥0.58 were independent risk factors for death in patients with cervical cancer ( OR = 1.857, 2.125, 2.337, 2.751, 2.457, 3.885 and 3.666; 95% CI 0.845 to 5.788, 0.726 to 5.924, 0.709 to 5.631, 0.693 to 5.727, 0.801 to 5.936, 1.244 to 6.423 and 1.031 to 5.612; P<0.01). Kaplan-Meier survival curve analysis result showed that the median survival time in cervical cancer patients with miR-34a expression level ≤0.39 and BMAL1 expression level ≥0.64 (21 cases) was significantly lower than that in the other cervical cancer patients (miR-34a expression level>0.39 or BMAL1 expression level<0.64, 55 cases): (26.4 ± 4.2) months vs. (34.2 ± 5.6) months, log-rank χ2 = 17.12, P<0.05. Conclusions:The expression level of serum miR-34a in patients with cervical cancer is significantly reduced and the expression level of BMAL1 is significantly increased, which is related to the condition, prognosis and HR-HPV infection. It can be used as a marker for the assessment of the condition and prognosis of cervical cancer. The combined detection of the two can significantly improve the sensitivity and specificity in predicting poor prognosis of cervical cancer.

Objective: To explore the effects of group sports game intervention on social ability and quality of life of children with austism spectrum disorders (ASD),so as to provide reference for rehabilitation intervention of social and quality of life of children with ASD. Methods: From September 2021 to January 2022, 72 children with ASD aged 4-6 in the children s rehabilitation department of Xiangyang Central Hospital were selected to participate in the study, and were randomly divided into experimental group ( n =36) and control group ( n =36). The control group received routine rehabilitation training (including individual sports game training), and the experimental group replaced individual sports game training with group sports game training on the basis of routine rehabilitation.The course content mainly included three parts: warm up before class, group sports games and relaxation after class. The course combined social skills with sports games, and was carried out in a group form (divided into 12 groups with 3 people in each group), and was trained five times a week for 60 minutes, for a total of 12 weeks. The scores of Childhood Autism Rating Scale (CARS), Autism Treatment Evaluation Checklist (ATEC), Social Responsiveness Scale (SRS) and Pediatric Quality of Life Inventory Version 4.0 Generic Core Scales (PedsQL 4.0) were observed before and after treatment. t-test and χ 2 test were used for statistical analysis. Results: There was no significant difference in SRS scores between the experimental group and the control group before intervention ( t =-0.63, P >0.05). After the intervention, the total response rate in the experimental group was 83.33 %, higher than 41.67% in the control group χ 2=13.33, P <0.05),and the SRS scores decreased in the experimental group and control group ( t =17.75,8.71, P <0.05). The SRS scale score of the experimental group Social perception (17.67±4.12) , social cognition (30.33±4.99) , social communication (50.33±9.39) , social motivation (24.25±6.78) scores and total scores ( 152.67± 25.82) were lower than those of the control group(22.17±5.34，36.00±4.13，62.58±11.07，34.42±7.13，186.33±29.03)( t = -4.88，-2.03，-2.13，-3.58，-3.01， P ＜0.05).After the intervention, the scores of social function (53.33±18.01) and total score (283.83±51.83) on PedsQL 4.0 scale in experimental group were higher than those in control group(23.33±15.13，218.00±39.01) ( t =4.42，3.52， P ＜0.05). After the intervention, Autism Treatment Evaluation Scale (ATEC) scores of experimental groups(44.33±14.72) was lower than that in control group ( 59.33±16.95)( t =-2.32, P ＜0.05). Conclusion The intervention of group sports game has a significant effect on improving social ability and life quality of children with ASD.