BACKGROUND:Due to the aging population and the increase in elderly diseases,research and utilization of epidermal stem cells in the elderly have become increasingly important.However,due to the weak proliferation and drying ability of epidermal stem cells in the elderly,their application in biomedical and clinical research is limited.OBJECTIVE:To investigate the ability of rotating cell culture system to promote the proliferation and maintenance of epidermal stem cells in elderly individuals.METHODS:Epidermal stem cells were obtained from children (7-12 years old) and the elderly (over 60 years old) using traditional planar culture methods.After identification by cell morphology and cell immunofluorescence,the differences in proliferation ability of epidermal stem cells between the children and the elderly were analyzed by flow cytometry,CCK-8 assay,and cell clone formation assay.Elderly epidermal stem cells were cultured using a rotating cell culture system and 3D TableTrix? microcarriers.The cell proliferation and expression of stemness markers were detected by Live/Dead staining,cell doubling efficiency,cell doubling time,and real-time quantitative polymerase chain reaction.RESULTS AND CONCLUSION:(1) Under planar culture conditions,the children group had stronger proliferative ability than the elderly group.(2) Compared with planar culture,the dynamic cultivation of elderly epidermal stem cells using a rotating cell culture system had a higher population doubling (P<0.01) and a shorter population doubling time (P<0.01).(3) The dynamic cultivation of rotating cell culture system promoted the self-assembly of elderly epidermal stem cells to form 3D human epidermal stem cell spheres based on microcarrier scaffolds,which had similar epidermal stem cell proliferation as the children group.(4) After dynamic culture,the expression of stemness marker genes (ITGA6 and ITGB1),basal cell marker genes (K5) and differentiation marker genes (K10 and K1) of elderly epidermal stem cells reached the same level as that of children.The results showed that the dynamic cultivation of rotating cell culture system can not only improve the efficiency of elderly epidermal stem cell culture,promote cell proliferation and self-assembly into 3D cell spheres,but also maintain a certain stemness of elderly epidermal stem cells without complete terminal differentiation.

BACKGROUND:Due to the aging population and the increase in elderly diseases,research and utilization of epidermal stem cells in the elderly have become increasingly important.However,due to the weak proliferation and drying ability of epidermal stem cells in the elderly,their application in biomedical and clinical research is limited.OBJECTIVE:To investigate the ability of rotating cell culture system to promote the proliferation and maintenance of epidermal stem cells in elderly individuals.METHODS:Epidermal stem cells were obtained from children (7-12 years old) and the elderly (over 60 years old) using traditional planar culture methods.After identification by cell morphology and cell immunofluorescence,the differences in proliferation ability of epidermal stem cells between the children and the elderly were analyzed by flow cytometry,CCK-8 assay,and cell clone formation assay.Elderly epidermal stem cells were cultured using a rotating cell culture system and 3D TableTrix? microcarriers.The cell proliferation and expression of stemness markers were detected by Live/Dead staining,cell doubling efficiency,cell doubling time,and real-time quantitative polymerase chain reaction.RESULTS AND CONCLUSION:(1) Under planar culture conditions,the children group had stronger proliferative ability than the elderly group.(2) Compared with planar culture,the dynamic cultivation of elderly epidermal stem cells using a rotating cell culture system had a higher population doubling (P<0.01) and a shorter population doubling time (P<0.01).(3) The dynamic cultivation of rotating cell culture system promoted the self-assembly of elderly epidermal stem cells to form 3D human epidermal stem cell spheres based on microcarrier scaffolds,which had similar epidermal stem cell proliferation as the children group.(4) After dynamic culture,the expression of stemness marker genes (ITGA6 and ITGB1),basal cell marker genes (K5) and differentiation marker genes (K10 and K1) of elderly epidermal stem cells reached the same level as that of children.The results showed that the dynamic cultivation of rotating cell culture system can not only improve the efficiency of elderly epidermal stem cell culture,promote cell proliferation and self-assembly into 3D cell spheres,but also maintain a certain stemness of elderly epidermal stem cells without complete terminal differentiation.

Objective:To summarize the key prenatal ultrasound diagnosis features of Apert syndrome, analyze the causes of missed diagnosis and misdiagnosis, and propose corresponding preventive strategies.Methods:A retrospective analysis was made on the medical records and prenatal ultrasound images of 15 fetuses (including 14 cases referred from other hospitals) who underwent prenatal ultrasound examination in Guangdong Women and Children Hospital from August 2014 to May 2022 and were eventually clinically confirmed as Apert syndrome by induction or after birth. By conducting a comparative analysis, particularly focusing on the initial and final diagnoses of referral cases, the key ultrasound diagnostic points of Apert syndrome and the causes for missed and misdiagnosis were summarized.Results:①Diagnostic accuracy: Among the 15 fetuses, 11 cases (73.3%) were correctly diagnosed by prenatal ultrasound and 4 cases (26.7%) were missed diagnosis and misdiagnosis. For the 14 referral cases, only 2 cases (14.3%) were correctly identified in the initial diagnosis at the referring hospital (14.3%), and 12 cases (85.7%) were missed missed diagnosis and misdiagnosis. ②Detection rate of ultrasound signs: In the 15 fetuses with Apert syndrome, the detection rate of " cloverleaf" skull was 13.3% (2/15), premature coronal suture was 66.7% (10/15), the " brain shadowing sign" and flat occiput were both 93.3% (14/15), prominent forehead, hypertelorism and bilateral syndactyly of hands were all 100% (15/15), and bilateral syndactyly of feet was 73.3% (11/15). ③Analysis of missed diagnosis and misdiagnosis: Among the 4 cases of missed diagnosis and misdiagnosis in our hospital, premature closure of coronal suture, " brain shadowing sign", flat occiput and hypertelorism were all not recognized. Among these, 3 cases also missed the prominent forehead, bilateral syndactyly of hands and feet. Additionally, 1 case of bilateral syndactyly of hands was misdiagnosed as partial absence of metacarpals and phalangess.Conclusions:In the prenatal ultrasound diagnosis of fetal Apert syndrome, the symmetric syndactyly of both hands serves as an extremely important diagnostic clue. The " cloverleaf" skull is not common. The premature closure of coronal suture as a direct diagnostic sign with a high detection rate, highlighting its significance in the diagnostic of Apert syndrome. Furthermore, the high detection rates of characteristic ultrasound features such as prominent forehead, flat occiput, " brain shadowing sign" and hypertelorism could help to improve the accuracy of prenatal ultrasound diagnosis for Apert syndrome and effectively reduce missed and misdiagnosis.

OBJECTIVE: Flavonoids are the bioactive compounds in safflower (Carthamus tinctorius), in which chalcone synthase (CHS) is the first limiting enzyme. However, it is unclear that which chalcone synthase genes (CHSs) are participated in flavonoids biosynthesis in C. tinctorius. In this study, the CHSs in the molecular characterization and enzyme activities were investigated. METHODS: Putative chalcone biosynthase genes were screened by the full-length transcriptome sequences data in C. tinctorius. Chalcone biosynthase genes in C. tinctorius (CtCHSs) were cloned from cDNA of flowers of C. tinctorius. The cloned gene sequences were analyzed by bioinformatics, and their expression patterns were analyzed by real-time PCR (RT-PCR). The protein of CtCHS in the development of flowers was detected by polyclonal antibody Western blot. A recombinant vector of CtCHS was constructed. The CtCHS recombinant protein was induced and purified to detect the enzyme reaction (catalyzing the reaction of p-coumaryl-CoA and malonyl-CoA to produce naringin chalcone). The reaction product was detected by HPLC and LC-MS. RESULTS: Two full-length CtCHS genes were successfully cloned from the flowers of safflower (CtCHS1 and CtCHS3), with gene lengths of 1525 bp and 1358 bp, respectively. RT-PCR analysis showed that both genes were highly expressed in the flowers, but the expression of CtCHS1 was higher than that of CtCHS3 at each developmental stage of the flowers. WB analysis showed that only CtCHS1 protein could be detected at each developmental stage of the flowers. HPLC and LC-MS analyses showed that CtCHS1 could catalyze the conversion of p-coumaryl-CoA and malonyl-CoA substrates to naringin chalcone. CONCLUSION CtCHS1 is involved in the biosynthesis of naringin chalcone in safflower.

This article reported a case of fetal giant hepatic hemangioma with cardiomegaly managed with intrauterine treatment. At 23 weeks of gestation, the patient was referred to Guangdong Women and Children Hospital due to abnormal abdominal echogenicity of the fetus, which was suspected to be a hepatic hemangioma or a hepatic arteriovenous fistula. The prenatal ultrasound at 26 weeks of gestation revealed an enlarged fetal hepatic hemangioma of 45 mm×35 mm×42 mm and an enlarged heart (cardiothoracic area ratio of 0.50). So, with the patient's informed consent, the fetus was treated with intrauterine administration of propranolol and dexamethasone and closely monitored by ultrasound. The volume of the lump still increased at the beginning of the medication, but started to shrink in the 7th week. Besides, the fetal cardiac load was reduced and the condition was controlled. The patient delivered at 37 weeks of gestation. The baby received a CT examination on the fourth day after birth which revealed an abdominal mass of 40 mm×30 mm×44 mm requiring no treatment, and no abnormalities were reported during a one-year follow-up.

OBJECTIVE：To investigate the correlation of storage life and effective composition content with color value of Carthamus tinctorius ，and to provide reference for the quality evaluation of C. tinctorius with different years of storage. METHODS：Using 24 batches of C. tinctorius from same place of production with different years of storage （0，1，2 years，8 batches each type ）as samples ，the contents of hydroxysafflor yellow A （HSYA）and kaempferol were determined by HPLC. Color value [lightness value （L*），red-green value （a*），yellow-blue value （b*）] were determined by spectrophotometer. SPSS 19.0 statistical software was used to analyze the correlation of storage life and effective composition content with color value. RESULTS：Kaempferol content was still high after 1 year or 2 years of storage （0.161％，0.061％，respectively）. However ，the content of HSYA decreased with the prolongation of the storage life （the average content of HSYA were 2.46％，1.58％，and 1.51％ after storage 0，1 and 2 years，respectively），and the color of the drug became darker （a* value decreased ）. Results of correlation analysis showed that the content of HSYA was positively associated with color value L*，a*（r＝0.430，0.781，P＜0.05 or P＜0.01）；the content of HSAY was negatively associated with storage life （r＝－0.777，P＜0.01）. There was no correlation between the remaining variables （P＞0.05）. CONCLUSIONS ：The longer the storage life ，the darker the color and the lower the content of HSYA ，so it is not suitable for over year and multiyear preservation.

Objective:To evaluate the effect and safety of adjustable closed loop(ACL) in the treatment of short bowel syndrome (SBS).Methods:From January 2017 to December 2017, the clinical data of 2 cases with ostomy in continuity after surgery in Children's Hospital Affiliated to Xi'an Jiaotong University were analyzed.The age was 1 and 3 months, both were females, and preservation of intestinal tube length were 65 cm and 60 cm respectively.They were all diagnosed SBS with a lot of stool like water.Self-made ACL was installed to maintain SBS.The blood circulation of stoma, weight, BMI, defecating, abdomen, nutrition status changes, nursing convenience, intestinal infection susceptibility were observed.Results:There was no necrosis in stoma, and the overall condition was improved, and no significant increase in abdominal distension.ACL used in babies with ostomy in continuity could improve the anus stool, weight, nursing convenience.Conclusion:ACL installed in baby with SBS could contribute to intestinal management, and is easy to install.

Objective To study on the factors of countercurrent occured in hysterosalpingography to improve the understanding of countercurrent.Methods 180 patients who underwent hysterosalpingography due to infertility were recruited,and 63 of them who were involved in countercurrent in the process of hysterosalpingography were analyzed statistically.Results The single factor analy-sis demonstrated that such four factors of primary/secondary infertility,menstrual clean days,tubal obstruction or not,and depth of cannula were associated with countercurrent,while Logistic regression analysis indicated that the factors of menstrual clean days, tubal obstruction or not,and depth of cannula during the hysterosalpingography operation were closely related.Conclusion Counter-current are caused by the comprehensive impact of several comprehensive factors like menstrual clean day,tubal obstruction or not, and cannula operation.Therefore adequate preparation should be made before and during the operation,to reduce the occurrences of countercurrent.

Objective To investigate the ability of T2 mapping in early detection of the change of cartilage collagen by quantitative analysis.Methods 20 porcine patellae were randomly assigned into 4 groups,3 treated groups and 1 control group.The samples of 3 treated groups were respectively immersed in PBS with 100 mg/100 mL,1 50 mg/100 mL and 200 mg/100 mL typeⅡ collagenase for 4 h,whereas control group samples in PBS for 4 h.T2 relaxation times of superficial,deep and full layers of cartilage and mean densi-ty after Van Gieson stain were measured.The correlation of T2 relaxation times and mean density were statistically analyzed.Results Gross pathology showed all articular cartilages maintained intact after treatment.T2 relaxation times of superficial and full layers and mean density increased significantly in any treated group in comparison with control group(P <0.05).Significant differences in T2 relaxation times of superficial layers and mean density were found between any two treated groups,whereas no difference of T2 re-laxation time of full layers was found between 100 mg/100 mL and 1 50 mg/100 mL treated groups(P >0.05).Pearson correlation analysis showed a close correlation(r=-0.837)between T2 relaxation times of full layers and mean density.Conclusion T2 map-ping can adequately detect the change of cartilage collagen in the early stage of osteoarthritis,and shows a good clinical application prospect.

BACKGROUND: Lung fibroblasts are believed to play an important role in lung tissue repair and regenerative process. The differentiation potential of lung fibroblasts is not known very well.OBJECTIVE: To investigate the multi-differentiation capacity of human lung fibroblasts.DESIGN: An observational comparative experiment.SETTING: Department of Respiratory Medicine, Beijing Shijitan Hospital and Central Laboratory of the First Hospital of Tsinghua University.MATERIALS: This study was performed at the Central Laboratory of the First Hospital of Tsinghua University from March 2006 to October 2006. Human adult lung fibroblasts were isolated as primary cultures from resected lung of patients with lung cancer.The study was approved by hospital's Medical Ethics Committee, and informed consent was signed. Dulbecco's modified Eagle's medium (DMEM), fetal calf serum (FCS), trypsin- ethylenediamine tetraacetic acid (EDTA), naphthol AS-MX phosphate and Fast Red TR were purchased from Sigma, USA. Mouse anti-human osteopondn antibody was from R&D Systems China Co., Ltd,Polyvinyl difluoride (PVDF) membranes were from Bio-Rad Laboratories Inc, Hercules, CA.METHODS: Human adult lung fibroblasts were isolated as primary cultures. Human lung fibroblasts were cultured in an osteogenic medium (containing 109-10-5 mol/L dexamethasone, 50 mg/L vitamin C and 10 mmol/L β-glycerophospate) and adipogenic medium (containing 15% horseurm, 10-8 mol/L dexamethasone and 10 mg/L insulin), or control medium (10% FCS).Ostcoblasts were detected by alkaline phosphatese (ALP) and calcium salt staining. The expression of osteopontin was measured by Western blotting. Oil Red-O staining was used for identification of mature adipocytes.MAIN OUTCOME MEASURES: The differentiation of lung fibroblasts induced by osteogenie medium; The differentiation of lung fibroblasts induced by adipogenic medium.RESULTS: With the induction of ostcogenic medium for 2 weeks, the differentiation of lung fibroblasts was induced by osteogenic and adipogenic medium, respectively. The expression of ALP and osteopontin was increased and the deposition of calcium salt was detected. After lung fibroblasts were cultured in adipogenic medium for 14 days, part of the cells gradually experienced morphological change from original spindle into oval shape. Oil Red -O staining indicated lipid drops accumulation within cytoplasm.CONCLUSION: Under certain condition, human long fibroblasts could differentiate into osteoblasts or adipocytes that were characterized by bone marrow mesenchymal stem cells.