1.Effect of Different Fermentation Conditions on Fungal Community and Chemical Composition of Aurantii Fructus
Zhihong YAN ; Xiumei LIU ; Qiuyan GUAN ; Yonggui SONG ; Zhifu AI ; Genhua ZHU ; Yuhui PING ; Ming YANG ; Qin ZHENG ; Huanhua XU ; Dan SU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):254-262
ObjectiveTo investigate the effects of different fermentation methods and times on the fungal flora and chemical composition of Aurantii Fructus, in order to obtain the optimal fermentation conditions and flora structure, and to ensure the stability and controllability of the fermented varieties. MethodsScanning electron microscopy was used to observe and analyze the colony characteristics on the surface of Aurantii Fructus under different fermentation conditions. Internal transcribed spacer 2(ITS2) high-throughput sequencing, combined with fungal community diversity analysis and fungal community structure analysis, were used to obtain the fungal flora microbial categories of Aurantii Fructus under the conditions of traditional pressure-shelf fermentation and non-pressure-shelf natural fermentation for 7, 14, 21 d(numbered Y1-Y3 for the former, and numbered F1-F3 for the latter), respectively. At the same time, the chemical components in the fermentation process were detected by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS), combined with principal component analysis(PCA), partial least squares-discriminant analysis(PLS-DA) and compound retention time, parent ions, characteristic fragment ions and other information, the differential compounds between the different fermentation samples were screened and identified. ResultsThe analysis of fungal community diversity showed that the dominant flora did not change at different fermentation time points in the traditional pressure-shelf fermentation method, while in the non-pressure-shelf natural fermentation method, there was a significant difference with the fermentation process, and at the genus level, the dominant genus of samples Y1, Y2, Y3 and F2 was Aspergillus, while the dominant genera of samples F1 and F3 were both Rhizopus. This indicated that the microbial growth environment provided by the traditional fermentation method was more stable, and the microbial community structure was more stable, which was more conducive to the stable and controllable fermentation process and fermented products. A total of 155 compounds were identified by compositional analysis, including 70 flavonoids, 38 coumarins, 10 alkaloids, 34 organic acids and 3 other compounds. After fermentation, two new components of ribalinine and pranferin were produced. Different fermentation conditions also brought about differences in chemical composition, multivariate statistical analysis obtained 26 differential compounds under two different fermentation methods, mainly including flavonoids, organic acids and coumarins. Comprehensively, the microbial community structure of samples fermented by the traditional pressure-shelf method of Aurantii Fructus for 14 d was stable, the species richness was high and the overall content of differential compounds was high, which was the optimal processing condition. ConclusionCompared with non-pressure-shelf natural fermentation, the traditional method has obvious advantages in terms of the stability of the microbial community structure and the content of chemical compounds, and the optimal condition is 14 days of fermentation. This study is helpful to promote the quality stability and fermentation bioavailability of fermented products of Aurantii Fructus, as well as to provide an experimental basis for the further improvement of the quality control methods of this variety.
2.Effect of Different Fermentation Conditions on Fungal Community and Chemical Composition of Aurantii Fructus
Zhihong YAN ; Xiumei LIU ; Qiuyan GUAN ; Yonggui SONG ; Zhifu AI ; Genhua ZHU ; Yuhui PING ; Ming YANG ; Qin ZHENG ; Huanhua XU ; Dan SU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):254-262
ObjectiveTo investigate the effects of different fermentation methods and times on the fungal flora and chemical composition of Aurantii Fructus, in order to obtain the optimal fermentation conditions and flora structure, and to ensure the stability and controllability of the fermented varieties. MethodsScanning electron microscopy was used to observe and analyze the colony characteristics on the surface of Aurantii Fructus under different fermentation conditions. Internal transcribed spacer 2(ITS2) high-throughput sequencing, combined with fungal community diversity analysis and fungal community structure analysis, were used to obtain the fungal flora microbial categories of Aurantii Fructus under the conditions of traditional pressure-shelf fermentation and non-pressure-shelf natural fermentation for 7, 14, 21 d(numbered Y1-Y3 for the former, and numbered F1-F3 for the latter), respectively. At the same time, the chemical components in the fermentation process were detected by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS), combined with principal component analysis(PCA), partial least squares-discriminant analysis(PLS-DA) and compound retention time, parent ions, characteristic fragment ions and other information, the differential compounds between the different fermentation samples were screened and identified. ResultsThe analysis of fungal community diversity showed that the dominant flora did not change at different fermentation time points in the traditional pressure-shelf fermentation method, while in the non-pressure-shelf natural fermentation method, there was a significant difference with the fermentation process, and at the genus level, the dominant genus of samples Y1, Y2, Y3 and F2 was Aspergillus, while the dominant genera of samples F1 and F3 were both Rhizopus. This indicated that the microbial growth environment provided by the traditional fermentation method was more stable, and the microbial community structure was more stable, which was more conducive to the stable and controllable fermentation process and fermented products. A total of 155 compounds were identified by compositional analysis, including 70 flavonoids, 38 coumarins, 10 alkaloids, 34 organic acids and 3 other compounds. After fermentation, two new components of ribalinine and pranferin were produced. Different fermentation conditions also brought about differences in chemical composition, multivariate statistical analysis obtained 26 differential compounds under two different fermentation methods, mainly including flavonoids, organic acids and coumarins. Comprehensively, the microbial community structure of samples fermented by the traditional pressure-shelf method of Aurantii Fructus for 14 d was stable, the species richness was high and the overall content of differential compounds was high, which was the optimal processing condition. ConclusionCompared with non-pressure-shelf natural fermentation, the traditional method has obvious advantages in terms of the stability of the microbial community structure and the content of chemical compounds, and the optimal condition is 14 days of fermentation. This study is helpful to promote the quality stability and fermentation bioavailability of fermented products of Aurantii Fructus, as well as to provide an experimental basis for the further improvement of the quality control methods of this variety.
3.Promotive effect of high expression of nerve growth factor in Schwan-like cells induced by adipose-derived stem cells on growth of rat dorsal root ganglion cell protrusion
Qinghua ZHU ; Bo YUAN ; Yilun WANG ; Miao REN ; Xiaofei LI ; Simiao WANG ; Zixuan ZHEN ; Xiumei FU
Journal of Jilin University(Medicine Edition) 2025;51(4):984-995
Objective:To discuss the promotive effect of nerve growth factor(NGF),which is highly expressed in the adipose-derived stem cell(ADSC)-induced Schwann-like cells(SCLCs),on the growth of dorsal root ganglion(DRG)cell processes in the rats,and to clarify its mechanism.Methods:The ADSCs were extracted from the epididymal adipose tissue of the SD rats,and their multidirectional differentiation potential was identified through osteogenic,adipogenic,and chondrogenic induction.The ADSCs were induced to differentiate into the SCLCs,and the expression levels of glial fibrillary acidic protein(GFAP)and S100 calcium-binding protein β(S100β)protein in the ADSCs and SCLCs were detected by immunofluorescence staining and Western blotting methods.The DRG cells were isolated and cultured,and immunofluorescence staining was used to detect the βⅢ-tubulin expression in the DRG cells for identification.The SCLCs were co-cultured with the DRG cells(co-culture group),the single-culture DRG cells were regared as DRG group and toluidine blue staining was used to observe and measure the length of DRG cell processes under the optical microscope in co-culture group and DRG group.Small interfering RNA(siRNA)transfection was used to knock down NGF,and plasmid transfection was used to over-express NGF.Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the NGF mRNA expression levels in the cells in various groups;enzyme-linked immunosorbent assay(ELISA)method was used to detect the NGF protein levels in the cell supernatants.The transfected SCLCs were co-cultured with DRG cells and divided into control group,siNC/vector group,NGF knockdown group(si-NGF group),and NGF over-expression group(oe-NGF group).The lengths of DRG cell processes in various groups were observed.Results:The primary ADSCs adhered within 24 h after seeding,with a small number of lipid droplets remaining.After 3 d of culture,the cells were mostly short spindle-shaped,fusiform,or polygonal,growing rapidly in a vortex pattern.After passaging,the cells exhibited a uniform morphology,appearing as long spindles arranged in a fish-school pattern.After 14 d of adipogenic induction,the cell morphology changed from spindle-shaped to flat-round,with translucent lipid droplets forming in the cytoplasm,which were stained red by Oil Red O.After 28 d of osteogenic induction,the cells appeared sand-like with blurred morphology,and calcified nodules were observed,which were stained red by Alizarin Red and deposited in the extracellular matrix.After 28 d of chondrogenic induction in a 3D culture system,millet-sized chondrogenic spheres formed.Frozen sections of the spheres were stained with Alcian Blue,and acidic mucopolysaccharides in the cartilage tissue were stained blue under the microscope.Under the fluorescence microscope,the third-passage purified ADSCs showed positive expression of CD29[fluorescein isothiocy anate(FITC)-labeled green fluorescence]and CD44(Cy3-labeled red fluorescence).The immunofluorescence staining results showed that GFAP was labeled with FITC(green fluorescence),and S100β was labeled with Cy3(red fluorescence).The Western blotting results showed that compared with ADSCs,the expression levels of S100β and GFAP proteins in the SCLCs were increased(P<0.05).The primary DRG cells began to adhere 6 h after conventional culture,and after 3 d,the cell bodies appeared round and bright,with two linear processes extending from them.Under fluorescence microscope,the cells positively expressed the neuron-specific marker βⅢ-tubulin,confirming that the isolated cells were DRG cells.Compared with the ADSCs,the NGF protein expression level in the SCLCs was increased(P<0.05).Compared with DRG group,the length of DRG cell processes in co-culture group was the highest when DRG cells and SCLCs were co-cultured at a 1∶2 ratio(P<0.05).The RT-qPCR results showed that compared with si-NC group,the expression levels of NGF mRNA in the cell supernatant in si-NGF-1,si-NGF-2,and si-NGF-3 groups were significantly decreased(P<0.05),with si-NGF-1 showing the highest knockdown efficiency,which was selected for subsequent experiments.The ELISA results showed that compared with si-NC group,the NGF levels in the cell supernatant of si-NGF-1,si-NGF-2,and si-NGF-3 groups were decreased(P<0.05).Compared with Vector group,the expression level of NGF mRNA and NGF protein level in the supernatant in oe-NGF group were increased(P<0.05).Compared with control group and siNC/vector group,the length of DRG cell processes in si-NGF group was decreased(P<0.05),while the length of DRG cell processes in oe-NGF group was increased(P<0.05).Conclusion:ADSCs can be directionally differentiated into SCLCs,and the differentiated cells highly express NGF.Knockdown or overexpression of NGF can affect the growth of DRG cell processes.
4.Protective effect of adipose-derived stem cells combined with acellular scaffolds on dorsal root ganglion in rats with sciatic nerve injury and its mechanism
Xiaomin YU ; Qinghua ZHU ; Yilun WANG ; Miao REN ; Zijia LIU ; Yongyi YU ; Yuanliang DU ; Donghui LIU ; Sen GUO ; Xiumei FU
Journal of Jilin University(Medicine Edition) 2025;51(6):1542-1550
Objective:To observe the effects of adipose-derived stem cells(ADSC)combined with acellular scaffold(AS)on the ultrastructure of dorsal root ganglion and the protein and mRNA expression levels of ciliary neurotrophic factor(CNTF),Janus kinase 2(JAK2),phosphorylated JAK2(p-JAK2),signal transducer and activator of transcription 3(STAT3)and phosphorylated STAT3(p-STAT3)in the rats with sciatic nerve injury(SNI),and to clarify the protective effect of ADSC combined with AS on dorsal root ganglion in the SNI rats and its possible mechanism.Methods:The rat ADSCs were isolated and cultured and their multidirectional differentiation potential was detected.The AS of rats was prepared,and ADSCs were injected into the AS to construct tissue-engineered nerve.A total of 36 rats were randomly divided into control group,model group,AS group,and ADSC+AS group.The rats in control group were routinely fed,and the rats in other groups were used to establish the SNI models by resecting 10 mm of right sciatic nerve.The rats in model group received no further treatment,while the rats in AS group and ADSC+AS group were bridged with AS and the constructed tissue-engineered nerve at the two ends of the injured nerve,respectively.At 6 weeks after surgery,transmission electron microscope was used to observe the ultrastructure of dorsal root ganglion of the rats in various groups;immunofluorescence method was used to detect the protein expression levels of CNTF,p-JAK2,and p-STAT3 in dorsal root ganglion of the rats;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the mRNA expression levels of CNTF,JAK2,and STAT3 in dorsal root ganglion of the rats in various groups.Results:After 7 d of primary ADSC culture,a large number of large and long spindle-shaped cells were observed under the inverted microscope,arranged in clusters or whirlpools;red lipid droplets were observed with oil red O staining under microscope,and calcified nodules were observed with Alizarin red staining under microscope,indicating that the isolated and cultured cells had multidirectional differentiation ability.Compared with normal nerve tissue,the level of DNA in AS of rats was significantly decreased(P<0.05).Compared with control group,the nuclear membrane of dorsal root ganglion cells in model group was uneven and serrated,the number of organelles in the cytoplasm was decreased,mitochondria were swollen with broken or missing cristae and unclear structure;the CNTF protein and mRNA expression levels were significantly decreased(P<0.01),the p-JAK2 and p-STAT3 protein expression levels were significantly increased(P<0.01),and the JAK2 and STAT3 mRNA expression levels were significantly increased(P<0.01).Compared with model group,the serrated change of nuclear membrane of the dorsal root ganglion cells in AS group was significantly alleviated,the number of organelles in the cytoplasm was increased,and mitochondrial swelling was reduced;in ADSC+AS group,the nuclear membrane of dorsal root ganglion cells tended to be intact,the number of organelles was increased,and mitochondrial swelling and vacuolization were significantly reduced;the CNTF protein and mRNA expression levels in the dorsal root ganglion in AS group and ADSC+AS group were significantly increased(P<0.01),the p-JAK2 and p-STAT3 protein expression levels were significantly decreased(P<0.01),and the JAK2 and STAT3 mRNA expression levels were significantly decreased(P<0.01).Compared with AS group,the CNTF protein and mRNA expression levels in ADSC+AS group were significantly increased(P<0.05 or P<0.01),the p-JAK2 and p-STAT3 protein expression levels were significantly decreased(P<0.01),and the JAK2 and STAT3 mRNA expression levels were significantly decreased(P<0.01).Conclusion:The application of ADSC combined with AS can improve the ultrastructure of dorsal root ganglion in the SNI rats,and the mechanism may be related to the increased CNTF expression and decreased activation of the JAK2/STAT3 signaling pathway in the dorsal root ganglion by ADSC combined with AS application.
5.Evaluation of the Applicability of the Dampness Syndrome Scale of Chinese Medicine(DSSCM)in Patients with Atopic Dermatitis
Li'an ZHU ; Liu ZHONG ; Yu ZHANG ; Siqi YE ; Xiumei MO
Journal of Guangzhou University of Traditional Chinese Medicine 2025;42(10):2367-2373
Objective To evaluate the applicability of the Dampness Syndrome Scale of Chinese Medicine(DSSCM)in patients with atopic dermatitis(AD).Methods A cross-sectional study was conducted 204 AD patients.Demographic data and scores of the DSSCM,Patient-Oriented Eczema Measure(POEM),Atopic Dermatitis Control Tool(ADCT),Scoring Atopic Dermatitis(SCORAD),Eczema Area and Severity Index(EASI),and Investigator's Global Assessment(IGA)were collected.SPSS and AMOS were used to analyze the reliability and validity of the DSSCM and then the correlation between dampness syndrome and clinical AD indicators was explored.Results The overall Cronbach's α coefficient of DSSCM was 0.90,and the Spearman-Brown split-half reliability coefficient was 0.81.Content validity results indicated that,except for items DSSCM 25,DSSCM 26,DSSCM 27,DSSCM 28,and DSSCM 29(with correlation coefficients<0.30),all other items showed correlation coefficients>0.40 with their respective dimensions and the total scale score.The success rates of convergent and discriminant validity tests exceeded 80%,and confirmatory factor analysis demonstrated good fit in the indices of x2/df and RMSEA.Correlation analysis revealed that DSSCM scores were significantly correlated with body mass index(BMI),Dermatology Life Quality Index(DLQI)scores,POEM scores,ADCT scores,pruritus scores,sleep scores,or SCORAD scores(P<0.05 or P<0.01).However,no significant correlations were observed between DSSCM scores and disease duration,age,IGA scores,or EASI scores(P>0.05).Conclusion The DSSCM demonstrates good reliability and validity in AD patients and its scores are correlated with clinical AD indicators,making it a suitable tool for assessing dampness syndrome in this population.
6.Mechanism of protection of motor neurons in spinal cord anterior horn of SNI rats by acellular nerve allografts via the Bcl-2/Cyt-C/Apaf-1 signalling pathway
Mengyuan Zheng ; Zitong Hao ; Qinghua Zhu ; Zhuangzhuang Tian ; Xingda Guo ; Yuhe Zheng ; Cheng Li ; Xiumei Fu
Acta Universitatis Medicinalis Anhui 2025;60(11):2035-2042
Objective:
To investigate the protective effects and mechanisms of acellular nerve allografts (ANA) on motor neurons in the spinal cord anterior horn of sciatic nerve injury ( SNI) rats .
Methods:
SPF grade male SD rats were randomly divided into normal , model , ANA-bridged (bridge group) , and autologous nerve transplantation groups (autograft group) , with 6 rats in each group . The SNI rat model was established using the right sciatic nerve clamp method for 10 mm . In the bridge group , the ANA was bridged to the two severed ends of the injured sciatic nerve , and in the autograft group , the autologous nerves were flipped head to tail and then bridged to the two se- vered ends . A spectrophotometer was applied to determine the DNA content in normal nerves and ANA . The foot- print test was used to determine the sciatic nerve function index (SFI) of the rats in each group , the wet weight ra- tio of the anterior tibialis muscle was calculated . The morphology and structure of the anterior horn motor neurons of the spinal cord of each group were observed by HE staining. The immunofluorescence and Western blot were used to detect Apaf-1 , Caspase-3 , Bcl-2 , Bax , and Cyt-C proteins expression in the L4-6 segment of the spinal cord .
Results:
The DNA content in the ANA prepared in this study was significantly lower than that in normal nerves (P < 0. 05) . Compared with the normal group , the SFI and wet weight ratio of the anterior tibialis muscle were re- duced in the model group (P < 0. 001) ; compared with the model group , both SFI and wet weight ratio of the ante- rior tibialis muscle significantly increased in the bridge group and the autografts group ( P < 0. 05 , P < 0. 001) , and the SFI and wet weight ratio of the anterior tibialis muscle in the autograft group were higher than those in the bridge group (P < 0. 001 , P < 0. 01) . The results of HE staining showed that the motor neurons in the anterior horn of the spinal cord of the normal group were structurally intact and had clear cytosolic boundaries; the neurons in the model group were lysed and necrotic , with blurred cytosolic boundaries; the neurons in the bridge group were less lysed and necrotic , but the nuclear translocation phenomenon could still be seen; the neurons in the autograft group were morphologically and structurally intact with clear cytosolic boundaries . Compared with the normal group , the expression of Apaf-1 , Caspase-3 , Bax and Cyt-C proteins significantly increased in the model group (P < 0. 001 , P < 0. 01 , P < 0. 01 , P < 0. 05) . Compared with the model group , the expression of Apaf-1 , Caspase- 3 , Bax , and Cyt-C proteins significantly decreased (P < 0. 001 , P < 0. 05 , P < 0. 05 , P < 0. 05) ; but the expres- sion of Bcl-2 protein significantly increased in the bridge group and the autograft group (P < 0. 05) . The expression of Apaf-1 , Caspase-3 , Bax and Cyt-C proteins in the autografts group was lower than that in the bridge group (P < 0. 001 , P < 0. 05 , P < 0. 05 , P < 0. 05) .
Conclusion
ANA can exert a protective effect on motor neurons in the anterior horn of the spinal cord of SNI rats by improving the morphology and structure of neurons , increasing the ex- pression of Bcl-2 protein , but decreasing the expression of Cyt-C , Bax , Caspase-3 , and Apaf-1 proteins in the spi- nal cord . The mechanism of ANA may be related to the Bcl-2/Cyt-C/Apaf-1-mediated mitochondrial apoptosis sig- naling pathway .
7.Rehabilitation effects of psychomotor therapy on young and middle-aged schizophrenic inpatients
Haiyun LI ; Liang MING ; Daojin WANG ; Wenli ZHU ; Xiumei WU ; Yuan PAN ; Tingting JIANG
Sichuan Mental Health 2024;37(2):114-119
BackgroundRehabilitation for schizophrenia typically relies on pharmacological interventions, yet their efficacy in improving social function and quality of life remains limited. In recent years, non-pharmacological approaches have shown promise in enhancing rehabilitation outcomes. However, research on the effectiveness of psychomotor therapy specifically for young and middle-aged schizophrenic inpatients is limited. ObjectiveTo explore the effects of psychomotor therapy on the rehabilitation of young and middle-aged schizophrenic inpatients, and to provide a reference for treatment strategies. MethodsA total of 104 young and middle-aged schizophrenic inpatients who met the International Classification of Diseases,tenth edition (ICD-10) diagnostic criteria and hospitalized in the Fourth People's Hospital of Wuhu from June 2021 to June 2022 were selected. Patients were randomly divided into two groups of 52 each using random number table method. Both groups received treatment with risperidone tablets (2~4 mg/d) , along with routine nursing care. Additionally, the research group received an extra 45~55 minutes of psychomotor therapy 2~3 times per week for 12 weeks. The control group received the same psychomotor therapy after the study. Positive and Negative Symptom Scale (PANSS), Scales of Social-skills for Psychiatric Inpatient (SSPI) and Insight and Treatment Attitude Questionnaire (ITAQ) were used to assess the patients before the intervention and at 4th, 8th and 12th week after the intervention. ResultsThe main effects of intervention at different time points for PANSS positive symptoms, negative symptoms and general psychopathology subscale scores, PANSS total score, SSPI score and ITAQ score were all statistically significant (F=33.989, 204.245, 82.817, 279.596, 26.144, 7.463, P<0.01). Furthermore, statistically significant between-group differences were observed in PANSS negative symptoms and general psychopathology subscale scores, PANSS total score, SSPI score and ITAQ score (F=30.053, 5.306, 33.417, 33.013, 18.608, P<0.05 or 0.01). Moreover, the interaction effect of time and group were statistically significant for PANSS positive symptoms, negative symptoms and general psychopathology subscale score, PANSS total score and SSPI score (F=3.472, 9.798, 3.843, 14.390, 20.661, P<0.05 or 0.01). After 12 weeks of intervention, the research group exhibited statistically significantly lower PANSS total score and subscale scores compared with baseline (P<0.01), while their SSPI total score was significantly higher than that of control group(P<0.01). Additionally, compared with the control group, the research group had statistically significantly lower PANSS total score and subscale scores , while their SSPI score was statistically significantly higher than those of control group(P<0.01). ConclusionPsychomotor therapy may contribute to the improvement of the psychiatric symptoms and social function in young and middle-aged inpatients with schizophrenia, enhancing their rehabilitation outcomes. [Funded by Health Commission Scientific Research Project of Wuhu (number, WHWJ2021y073)]
8.Effect of adipose-derived stem cell-derived exosomes on migration ability of macrophages in vitro
Bo YUAN ; Jiayi XIE ; Siyu JIANG ; Yajun MENG ; Qinghua ZHU ; Xiaofei LI ; Xiumei FU ; Lide XIE
Journal of Jilin University(Medicine Edition) 2024;50(3):718-727
Objective:To discuss the effect of adipose-derived stem cell-derived exosomes(ADSC-Exos)on the migration ability of the macrophages RAW264.7,and to clarify its role in promoting function of the macrophages.Methods:The adipose tissue adjacent to epididymis of the SD rats was isolated to perform primary culture of the adipose-derived stem cells(ADSCs).The adipogenic and osteogenic differentiation induction was conducted,and the multidirectional differentiation potential of the ADSCs was detected by oil Red O and Alizarin red staining.Western blotting and immunofluorescence methods were used to detect the positive expressions of the ADSCs markers CD29 and CD44;the ADSC-Exos were extracted by Exos isolation kit,and the morphology,size,and distribution of particle size of the ADSC-Exos were examined by transmission electron microscope and nanoparticle tracking analyzer;the expression levels of exosome-specific markers CD9 and TSG101 proteins in the ADSC-Exos were detected by Western blotting method;the uptake of ADSC-Exos by the macrophages was observed by tracing method.The macrophages RAW264.7 were divided into control group,10 mg·L-1 ADSC-Exos group,20 mg·L-1 ADSC-Exos group,and 40 mg·L-1 ADSC-Exos group.The activities of the macrophages in various groups were detected by 5-ethynyl-2'-deoxyuridine(EdU)staining;the number of migration macrophages in various groups was detected by Transwell chamber assay;the adhesion of macrophages in various groups was observed by fluorescence microscope.Results:After 24 h of primary culture,the ADSCs adhered to the wall and exhibited scattered,elongated shapes;after 7 d of culture,the adherent cells showed a comb-like,vortex-like orderly arrangement,resembling fibroblasts;after 10 passages,the irregular morphology of the ADSCs and decreased proliferation rate were found.The isolated ADSCs showed potential for the osteogenic and adipogenic differentiation,and the expressions of CD29 and CD44 proteins were positive.The transmission electron microscope observation resuls showed that the ADSC-Exos appeared disc-shaped,and the main peak of particle size distribution was around 132 nm.The CD9 and TSG101 proteins were positively expressed in the ADSC-Exos,indicating successful extraction.The fluorescence microscope results showed red fluorescence signals around the nuclei of the RAW264.7 cells,indicating the uptake of ADSC-Exos by the macrophages.Compared with control group,the rates of EdU positive cells in 10,20,and 40 mg·L-1 ADSC-Exos groups were significantly increased(P<0.05);compared with 10 mg·L-1 ADSC-Exos group,the rate of EdU positive cells in 20 mg·L-1 ADSC-Exos group was significantly increased(P<0.05).Compared with control group,the numbers of migration cells in 10,20,and 40 mg·L-1 ADSC-Exos groups were significantly increased(P<0.05);compared with 10 mg·L-1 ADSC-Exos group,the numbers of migration cells in 20 and 40 mg·L-1 ADSC-Exos groups were significantly increased(P<0.05).Compared with control group,the numbers of the adherent macrophages in 10,20,and 40 mg·L-1 ADSC-Exos groups were significantly increased(P<0.05);compared with 10 mg·L-1 ADSC-Exos group,the number of adherent macrophages in 20 mg·L-1 ADSC-Exos group was significantly increased(P<0.05).Conclusion:The ADSC-Exos can be internalized by the macrophages and they can enhance the migration ability of the macrophages by affecting the cell adhesion.
9.Analysis of pathogenic bacteria of urinary tract infection in patients with mental disorders of different genders
Lei YUE ; Xiumei ZHU ; Shijing HE ; Shaojing YAN
Sichuan Mental Health 2022;35(3):245-249
ObjectiveTo analyze the gender differences in the pathogen distribution and drug susceptibility of bacteria causing urinary tract infection among psychiatric inpatients in a hospital in Guangzhou, and to provide a basis for clinical diagnosis and rational use of drugs in treatment. MethodsClinical data of 326 psychiatric patients complicating urinary tract infection in a hospital in Guangzhou from 2019 to 2020 were retrospectively analyzed, including 126 males and 200 females. Data including gender, age, identification results of urinary tract pathogens from urine samples and drug susceptibility results were collected. The differences in bacterial distribution and drug resistance rate of urinary tract infection pathogens in patients of different genders were analyzed. ResultsA total of 326 strains of urinary tract infection bacteria were isolated, including 103 strains (31.60%) of multi-drug resistant bacteria. Male and female urinary tract infection in patients with multi-drug resistant bacteria were detected 52 strains (41.27%) and 51 strains (25.50%), the detection rate of multi-drug resistant bacteria in female patients was significantly higher than that in male patients, with statistical difference (χ2=8.895, P<0.01). In terms of bacterial distribution, the composition ratio of Escherichia coli in female patients was higher than that in male patients (χ2=14.794), while the composition ratio of Acinetobacter baumannii and Pseudomonas aeruginosa was lower than that in male patients (χ2=13.665, 4.054), with statistical difference (P<0.05 or 0.01). The drug susceptibility results showed that Escherichia coli isolated from female patients were less resistant to various antibiotics such as ampicillin/sulbactam, aztreonam, cefepime, ceftazidime, levofloxacin, imipenem and meropenem than those from male patients (χ2=5.028~17.680, P<0.05 or 0.01). ConclusionThe prevalence rate and bacterial distribution of psychiatric patients complicating urinary tract infection differ between patients of different genders, furthermore, the rate of drug resistance for Escherichia coli is lower in female patients than that in male patients.
10.Predictive value of transvaginal ultrasound measurement of cervical length in first and second trimester on spontaneous preterm birth in singleton pregnancies
Qian WU ; Junya CHEN ; Xiaoxiao ZHANG ; Lixin FAN ; Yuchun ZHU ; Baihua JING ; Linlin WANG ; Ruina HUANG ; Chen LI ; Xiumei LI ; Huixia YANG
Chinese Journal of Perinatal Medicine 2022;25(5):326-331
Objective:To explore the predictive value of transvaginal ultrasound measurement of cervical length (CL) in the first and second trimester on spontaneous preterm birth in singleton pregnant women.Methods:This study retrospectively recruited 2 254 singleton pregnancies without severe comorbidities at Peking University First Hospital from January 2019 to June 2019. CL was measured for all subjects using transvaginal ultrasound in the first (11-13 +6 weeks) and second trimester (21-23 +6 weeks). Differences in CL between women with preterm (preterm group) and full-term delivery (full-term group) as well as the CL during the first and second trimester were compared. The independent risk factors for preterm birth and the predictive value of CL in the first and second trimester for spontaneous preterm birth were also explored. Fisher's exact test, t-test, χ2 test, and logistic regression analysis, etc, were adopted for statistical analysis. Results:(1) For the 2 254 subjects, CL measured in the first trimester and second trimester were (36.1±4.2) mm (22.4-52.6 mm) and (36.9±5.3) mm (2.9-59.7 mm), respectively. The incidence of short cervix in the first trimester and second trimester were 0.31% (7/2 254) and 1.46% (33/2 254), respectively. When CL was ≤25.0 mm ( OR=43.92, 95% CI:6.83-282.49) or >25.0-≤30.3 mm ( OR=6.59, 95% CI:1.97-22.0) in the first trimester, the risk of short cervix increased in the second trimester (both P<0.05). (2) The total incidence of preterm delivery was 3.06% (69/2 254). CL and the incidence of short cervix did not differ significantly in the first trimester between the preterm and full-term group [(35.2±4.5) and (36.1±4.1) mm, t=-1.78, P=0.076; 1.5% (1/69) and 0.3% (6/2 185), χ 2=2.98, P=0.084]. Compared with the full-term group, CL was shorter and the incidence of short cervix was higher in the second trimester in the preterm group [(33.6±6.7) vs (37.0±5.2) mm, t=-5.12;8.7% (6/69) vs 1.2% (27/2 185), χ 2=25.80, P<0.001]. (3) Multivariate regression analysis showed that age ≥35 years ( OR=2.05, 95% CI:1.22-3.46), history of spontaneous preterm birth ( OR=25.25, 95% CI:5.01-127.28), conception assisted by reproductive technology ( OR=10.39, 95% CI:2.39-50.33), and short cervix during the second trimester were independent risk factors for premature delivery. (4) There was no significant difference in the risk of preterm delivery when comparing to those with CL≤25.0 mm, >25.0-≤30.3 mm, >30.3-≤33.0 mm, >33.0-≤35.7 mm, >35.7-≤38.7 mm women with CL>38.7 mm during the first trimester (all P>0.05). The risk of premature delivery was relatively increased for those with CL≤25.0 mm,>25.0-≤29.5 mm, >29.5-≤33.6 mm, >33.6~≤36.8 mm, >36.8~≤40.1 mm during the second trimester compared to those with CL>40.1 mm [ OR (95% CI):17.64 (4.99-62.32), 6.89 (2.11-22.55), 3.58 (1.34-9.59), 4.04 (1.58-10.32), 3.34 (1.28-8.67), respectively , all P<0.05]. (5) When CL≤25.0 mm and ≤29.5 mm in the second trimester were used as the cut-off value, the prediction of preterm delivery was with a sensitivity of 8.70% and 17.39%, specificity of 98.80% and 95.29%, positive predictive value of 18.20% and 10.43%, negative predictive value of 97.16% and 97.34%, and the accuracy rate of 96.01% and 92.90%, respectively. Conclusions:There were no significant differences in CL and the incidence of short cervix during the first trimester among women with preterm or full-term delivery. CL in the first trimester is not an independent risk factor for preterm birth, but the risk of short cervix in the second trimester is increased when CL≤30.3 mm in the first trimester. The shorter the cervix during the second trimester, the greater the risk of preterm birth.


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