ObjectiveThis study aims to investigate the effects of different sizes of seedlings and melatonin treatment on physiological and biochemical indicators and bolting-related gene expression in Angelica sinensis， find substances related to early bolting， and elucidate the inhibitory mechanism of melatonin on bolting. MethodsSpectrophotometry was used to detect the related enzyme activities of A. sinensis leaves. The contents of endogenous hormones and polyamines were detected using ultra-high performance liquid chromatography-tandem mass spectrometry. Real-time polymerase chain reaction （Real-time PCR） was used to detect the expression levels of bolting-related genes. Inter-group differential indicator analysis， orthogonal partial least squares discriminant analysis， and principal component analysis were comprehensively applied to identify factors related to early bolting. ResultsEndogenous jasmonic acid and melatonin were identified as the most important factors affecting early bolting. Secondly， the activity of antioxidant enzymes， abscisic acid content， gibberellin content， and the expression levels of CO3， HD3A， and FD genes had important effects on the bolting process. Compared with small seedlings， exogenous melatonin treatment mainly inhibited early bolting by increasing endogenous melatonin content， reducing gibberellin content， and decreasing the expression levels of SOC1 and FD genes. ConclusionExogenous melatonin can inhibit early bolting in A. sinensis by regulating its physiological， biochemical， and gene expression levels.

ObjectiveThis study aims to investigate the effects of different sizes of seedlings and melatonin treatment on physiological and biochemical indicators and bolting-related gene expression in Angelica sinensis， find substances related to early bolting， and elucidate the inhibitory mechanism of melatonin on bolting. MethodsSpectrophotometry was used to detect the related enzyme activities of A. sinensis leaves. The contents of endogenous hormones and polyamines were detected using ultra-high performance liquid chromatography-tandem mass spectrometry. Real-time polymerase chain reaction （Real-time PCR） was used to detect the expression levels of bolting-related genes. Inter-group differential indicator analysis， orthogonal partial least squares discriminant analysis， and principal component analysis were comprehensively applied to identify factors related to early bolting. ResultsEndogenous jasmonic acid and melatonin were identified as the most important factors affecting early bolting. Secondly， the activity of antioxidant enzymes， abscisic acid content， gibberellin content， and the expression levels of CO3， HD3A， and FD genes had important effects on the bolting process. Compared with small seedlings， exogenous melatonin treatment mainly inhibited early bolting by increasing endogenous melatonin content， reducing gibberellin content， and decreasing the expression levels of SOC1 and FD genes. ConclusionExogenous melatonin can inhibit early bolting in A. sinensis by regulating its physiological， biochemical， and gene expression levels.

Objective To detect diatoms in tissues and water samples from drowning cases by gene array,and explore the application value of DNA microarray.Methods Diatoms in the lung,liver,kidney samples,and on-site water samples from 19 drowned corpses that appeared in the rivers of Zhongshan City between July 2021 and April 2022,were detected by gene arrays.Moreover,diatom detection was also performed on 28 samples from 7 corpses by microscope.Then the testing results were compared and analyzed.Results For those 28 samples,diatom types in both tissue and water samples by gene detection were more than those detected by microscopy,the detection was statistically significant(P<0.05).However,there was no statistically significant difference(P>0.05)in the detection of diatom types between tissue samples and water samples both using gene arrays.In terms of tissue samples,diatoms were easier to be detected in lung than liver or kidney,and there were also more types of diatoms detected in lung.Diatom detection by gene arrays showed that the diatom types showed a decreasing trend as the tissue was further away from the entrance of body,and the difference of diatom types increased among tissues.Conclusion The gene array can effectively detect the diatom types and distribution characteristics in drowned corpses,and has great potential in the research of the traceability analysis of corpses found in water and the causes of death.

Objective To investigate the effect of peptidyl arginine deiminase 4(PAD4)on the differentiation of mesenchymal stem cells isolated from oral bones(OMSC)and craniomaxillofacial development.Methods Immunofluorescence was used to detect the ex-pression of PAD4 in the mandibular of mice E13.5 embryo.A peptidyl arginine deiminase 4 knockout(PAD4-KO)mouse model was constructed.Craniomaxillofacial development was investigated by micro-CT.CCK-8 assay and Transwell assay were used to detect the OMSC proliferation ability and migration ability of PAD4-KO and wild type(WT)mouse.ALP staining was used to detect the changes in OMSC osteogenic differentiation ability.The expression of osteogenesis-related genes was detected by immunofluorescence and PCR assay.Results PAD4 was highly expressed in the mandibular tissue of mouse embryos at E13.5.On the cellular level,PAD4 was ex-pressed in the nucleus and mitochondria of OMSC.Compared to the WT mice,micro-CT showed that PAD4-KO mice had retrusive jaw and decreased mineralization.The proliferation and migration ability of OMSC in PAD4-KO mice were decreased.OMSCs lacking PAD4 had significantly decreased ALP staining level,and the expression levels of osteogenesis-related genes were decreased.In addition,it was found that PAD4 might affect OMSC mineralization by regulating Runx2 transcription.Conclusion PAD4 is expressed in the jaw during embryonic development.It might affect the embryonic development by regulating the proliferation and differentiation of OMSC,leading to craniomaxillofacial abnormalities

Objective To detect diatoms in tissues and water samples from drowning cases by gene array,and explore the application value of DNA microarray.Methods Diatoms in the lung,liver,kidney samples,and on-site water samples from 19 drowned corpses that appeared in the rivers of Zhongshan City between July 2021 and April 2022,were detected by gene arrays.Moreover,diatom detection was also performed on 28 samples from 7 corpses by microscope.Then the testing results were compared and analyzed.Results For those 28 samples,diatom types in both tissue and water samples by gene detection were more than those detected by microscopy,the detection was statistically significant(P<0.05).However,there was no statistically significant difference(P>0.05)in the detection of diatom types between tissue samples and water samples both using gene arrays.In terms of tissue samples,diatoms were easier to be detected in lung than liver or kidney,and there were also more types of diatoms detected in lung.Diatom detection by gene arrays showed that the diatom types showed a decreasing trend as the tissue was further away from the entrance of body,and the difference of diatom types increased among tissues.Conclusion The gene array can effectively detect the diatom types and distribution characteristics in drowned corpses,and has great potential in the research of the traceability analysis of corpses found in water and the causes of death.

Objective To investigate the effect of peptidyl arginine deiminase 4(PAD4)on the differentiation of mesenchymal stem cells isolated from oral bones(OMSC)and craniomaxillofacial development.Methods Immunofluorescence was used to detect the ex-pression of PAD4 in the mandibular of mice E13.5 embryo.A peptidyl arginine deiminase 4 knockout(PAD4-KO)mouse model was constructed.Craniomaxillofacial development was investigated by micro-CT.CCK-8 assay and Transwell assay were used to detect the OMSC proliferation ability and migration ability of PAD4-KO and wild type(WT)mouse.ALP staining was used to detect the changes in OMSC osteogenic differentiation ability.The expression of osteogenesis-related genes was detected by immunofluorescence and PCR assay.Results PAD4 was highly expressed in the mandibular tissue of mouse embryos at E13.5.On the cellular level,PAD4 was ex-pressed in the nucleus and mitochondria of OMSC.Compared to the WT mice,micro-CT showed that PAD4-KO mice had retrusive jaw and decreased mineralization.The proliferation and migration ability of OMSC in PAD4-KO mice were decreased.OMSCs lacking PAD4 had significantly decreased ALP staining level,and the expression levels of osteogenesis-related genes were decreased.In addition,it was found that PAD4 might affect OMSC mineralization by regulating Runx2 transcription.Conclusion PAD4 is expressed in the jaw during embryonic development.It might affect the embryonic development by regulating the proliferation and differentiation of OMSC,leading to craniomaxillofacial abnormalities

Objective To explore the inter-rater and within-rater reliability of the 30 categories of the International Classification of Functioning Disability,and Health(ICF)dysfunction set(ICF generic-30 set)in clinical function assessment,to provide a basis for its clinical application.Methods Ninety patients from the Neurorehabilitation Department of the First People's Hospital of Suqian City were enrolled in this study.All patients were evaluated by the same functional evaluator's ICF dysfunction combination at the two time points of admission and discharge for the internal reliability analysis of the evaluator.Thirty patients in the neurorehabilitation department were randomly selected to be evaluated by two ICF function evaluators at the same time to conduct test-retest reliability analysis among evaluators.The internal consistency of the Cronbach's α coefficient test scale,and the intra-group correlation coefficient(ICC)test inter-rater reliability and within-rater reliability.Results The Cronbach's α coefficients of the combination of ICF dysfunction,physical function and activity,and participation in the two subscales were 0.941,0.717,and 0.942,respectively.The inter-rater reliability analysis showed that the ICC of the total score of the scale and the total scores of physical function and activity and participation in the two subscales were 0.998,0.971,0.999(P<0.01),and the inter-rater reliability good;the internal reliability analysis of the evaluator showed that the total score of the scale and the total scores of physical function and activity and participation in the two subscales were 0.988,0.917,0.991(P<0.01),except that b640 sexual function is not applicable,and b130 energy Harmony drive function ICC is 0.558,b280 pain ICC is 0.409,ICC of other combination items is 0.838-0.986(P<0.01),the evaluator's internal reliability is good;90 patients'ICF dysfunction combination scale has 30 categories.The comparison between admission and discharge was statistically significant except that b640 was not applicable for sexual function(P<0.05).Conclusion The combination of ICF dysfunction has good internal consistency and inter-evaluator and intra-evaluator reliability in the functional evaluation of patients in neurorehabilitation department.

Objective:To evaluate the correlation between vascular hyperintensity of magnetic resonance fluid-attenuated inversion recovery (FLAIR) sequence(FVH) and related parameters of magnetic resonance perfusion weighted imaging (MR-PWI) in patients with middle cerebral artery stenosis cerebral infarction, and to explore the hemodynamic factors related to FVH and the effect of FVH on the short-term clinical prognosis of patients.Methods:A total of 116 patients with middle cerebral artery stenosis cerebral infarction in the Department of Neurology, Qingpu Branch of Zhongshan Hospital Affiliated to Fudan University from January 2020 to December 2022 were collected.According to the diagnostic criteria of FVH, they were divided into FVH (+ ) group (78 cases) and FVH(-) group (38 cases). All patients underwent magnetic resonance(MR) and MR-PWI scans.Demographic and cerebrovascular risk factors were collected, clinical neurological function of patients was assessed by national institate of health stroke scale(NIHSS) upon admission and discharge, and cognitive function of patients was assessed by mini-mental state examination (MMSE). Short-term clinical outcome was assessed using modified Rankin scale(mRS) at the 90th day after discharge.The degree of middle cerebral artery stenosis, positive or negative FVH, FVH score, hypoperfusion volume and MR-PWI related parameters, including peak time (Tmax), mean transit time (MTT), cerebral blood volume (CBV) and cerebral blood flow (CBF), were evaluated in relation to clinical symptoms.SPSS 22.0 statistical software was used for t test, Chi-square test and Pearson correlation analysis. Results:There were significant differences in hypoperfusion volume, Tmax, MTT and CBF between FVH (+ ) group and FVH(-) group( t=1.989, 3.830, 5.223, 3.911, all P<0.05). In terms of short-term clinical outcome, the improvement rate of neurological function ((8.25±6.39)%, (12.22±6.08)%) and MMSE score(25.48±1.59), (26.31±1.26) in FVH (+ ) group were significantly lower than those in FVH(-) group, and the number of patients with progressive stroke during hospitalization in FVH(+ ) group was more than that of FVH(-) group(22(28.21%), 4(10.53%)) (all P<0.05). Pearson correlation analysis showed that FVH score was positively correlated with hypoperfusion volume ( r=0.786, P<0.01) and MTT ( r=0.692, P<0.01), and negatively correlated with CBF ( r=-0.568, P<0.01), but no significant correlation with the degree of arterial stenosis ( r=0.363, P>0.05). Conclusion:FVH is closely related to the Tmax, MTT and CBF values shown in MR-PWI, and the incidence of stroke in progression and short-term adverse prognosis are more likely in FVH(+ ) group, suggesting that FVH can be used as a convenient imaging indicator to reflect the hypoperfusion status of patients with middle cerebral artery stenosis cerebral infarction, and can provide an objective basis for further individualized treatment.

Objective:To observe any effect of using the International Classification of Functioning, Disability and Health (ICF) rehabilitation set in rehabilitating the physical ability of hemiplegic stroke survivors.Methods:Ninety stroke survivors with hemiplegia were selected and randomly divided into an observation group ( n=45) and a control group ( n=45). The observation group was first assessed using the ICF and then given 60 minutes of individualized rehabilitation, 5 or 6 times a week for 4 weeks in addition to a weekly one-hour rehabilitation course. The control group were evaluated and treated traditionally. Before and after the intervention, both groups were evaluated using the Fugl-Meyer exercise assessment (FM) and functional independence assessment (FIM). Results:After the treatment the average upper and lower limb scores as well as the total FM scores had improved significantly. Those of the observation group were then significantly different from the control group′s averages.Conclusion:Basing rehabilitation on the ICF can more effectively improve the physical functioning of hemiplegic stroke survivors.

Objective:To evaluate the efficacy and safety of azacitidine combined with HAG regimen in the treatment of newly diagnosed elderly acute myeloid leukemia (AML) patients ineligible for intensive chemotherapy.Methods:Eighteen newly diagnosed elderly AML patients ineligible for intensive chemotherapy from July 2019 to September 2021 in the Second People's Hospital of Shenzhen were prospectively enrolled in this study. They were non-randomly divided into azacitidine combined with HAG regimen (AZA-HAG) group (9 cases) and decitabine combined with HAG regimen (DEC-HAG) group (9 cases). The primary endpoint of the study was overall response [complete remission (CR)+partial remission], and the secondary endpoints included CR + complete remission with incomplete count recovery (CRi), overall survival (OS) and drug safety. Kaplan-Meier method was used to analyze the OS.Results:The median age of 18 patients was 67 years old (60-77 years old) , and 8 of them were in high-risk group. After one course of treatment, the overall response and CR+CRi were observed in 7 of 9 patients in AZA-HAG group, and they were observed in 8 of 9 patients in DEC-HAG group, and there was no significant difference between the two groups (both P = 1.000). The median duration of CR+CRi was 7 months in both groups, and the median OS time was 12 months in both groups; there was no significant difference in OS between the two groups ( χ2 = 0.02, P = 0.895). In AZA-HAG group, 1 patient with TP53 mutation and 1 patient with ASXL1+RUNX1 mutation acquired CR, and 1 patient with NPM1 wild-type combined with FLT3-ITD and ASXL1 mutation did not respond. There was no significant difference in the incidence of grade 3-4 hematological adverse reactions between the two groups (all P < 0.05). Conclusions:Azacitidine combined with low-dose HAG regimen in the treatment of newly diagnosed elderly AML patients ineligible for intensive chemotherapy has satisfactory efficacy and long-term survival, and the adverse reactions can be tolerated.