| Part I The blood biochemical results of children with Crohn’s disease predicted the positive activity of the disease under endoscopeObjective: To explore the correlation between blood biochemical results and disease activity of children with CD under endoscopy,with a view to guiding clinical diagnosis and treatment.Methods: Retrospective analysis of children with Crohn’s disease diagnosed in our hospital from 2012.01 to 2019.04,according to the PCDAI clinical severity score,divided into two groups of light active period and moderate/severe active period.Clinical manifestations,blood biochemical indicators and treatment were analyzed.Results: Through general situation analysis,it was found that there were no statistical differences in gender,age and initial weight of children with mild and moderate to severe activity,but children with moderate to severe activity were significantly longer than those with mild activity in disease course(P=0.03);We analyzed the major clinical manifestations and found that abdominal pain,diarrhea,and weight loss are the major clinical manifestations of celiac disease in children.In addition,the children in the moderately and severely active group had a statistically significant difference in weight loss compared with those in the mildly active group(P<0.01);analysis of common clinical blood biochemical indicators revealed that the albumin in the moderately and severely active group was(37.34±5.65 vs.33.00±6.23,P=0.022),hemoglobin(107.00±14.89 vs.92.04±17.94,P=0.007),RBC(4.38±0.53 vs.3.94±0.73,P=0.041)levels decreased significantly compared with the lightly active group However,platelets(387.59±130.13 vs.492.75±132.77,P=0.016)and erythrocyte sedimentation rate(44.47±24.52 vs.75.50±36.36,P=0.005)were significantly higher than those in the mild group.Further,the analysis of endoscopic lesions,mucosa and pathological manifestations found that compared with children with mild activity,moderate to severe children are more likely to have total colon involvement,rectal involvement is also more common,and pathological manifestations are more It is easy to find manifestations such as ulcers,cobblestone signs,segmental lesions,granuloma,and eosinophil infiltration;there is no significant difference between the two groups of children in the treatment plan;and PCDAI,SES-CD and clinical blood biochemical indicators It was found that PCDAI had high accuracy in predicting endoscopic activity in children with CD.SES-CD was moderately correlated with PCDAI and Hb,and lowly correlated with RBC,ALB and ESR.Conclusion: Abdominal pain,diarrhea,and weight loss are the main clinical manifestations in children with Crohn’s disease.Weight loss,ALB,Hb,RBC,PLT,and ESR levels have important significance in the classification of disease activity.A sensitive indicator of lower activity.Part II Effect and mechanism of h UC-MSCs on nerve repair in HIBD rats under different neural differentiation induction statusObjective: Transplantation of human umbilical cord mesenchymal stem cells(h UC-MSCs)can promote hypoxic-ischemic brain damage(HIBD)nerve repair,but finding suitable seed cell to optimize transplantation and improve treatment efficiency is an urgent problem to be solved.Methods:(1)h UC-MSCs was induced to neural differentiation,dedifferentiation,and redifferentiation,observe the cell morphology under an inverted microscope,and analyze its gene expression;the HIBD rat model was established,and on the fifth day after the successful modeling,the cells of the transplanted group(h UC-MSCs,Dif,De-Dif,Re-Dif)were injected into the lateral ventricle of the brain with a stereotype,and the four groups of rats were tested by water maze and object-in-place behavioral experiments to test their learning and memory functions and their ability to explore new things.h UC-MSCs and De-Dif were subjected to GO analysis and KEGG pathway analysis;the differential genes and RARs genes of the two groups of cells were analyzed.(2)Based on previous studies,the cell lines of h SDF-1α,both of which were underexpressed(sih SDF-1)and overexpressed(adh SDF-1),were successfully established and detected by Western Blot.Results:(1)Normal h UC-MSCs,Dif,and De-Dif adherent cells showed flat spindle shape or slender needle tip shape,and the cells grew rapidly in a vortex shape.Re-Dif showed obvious neuron-like cell morphology.(2)Through the clustering analysis of the whole gene expression,the h UC-MSCs and De-Dif had the same high expression and low expression,and the gene expression pattern was the most similar.(3)In Morris water maze experiment,Sham(n=10),PBS(n=9),h UC-MSCs(n=9)and De-Dif(n=9)were transplanted into the HIBD model rats to test their effects on the learning and memory function of the HIBD rats.On the first day,there was no significant difference in the path length or escape latency of the four groups of HIBD rats,P>0.05.Hidden platform training on days 2-5,the De-Dif group took the shortest time to find the platform,and the difference was statistically significant(P=0.000,De-Dif vs.h UC-MSCs).On the 6th day of the platform exploration test,the frequency of the discovery platform in the De-Dif group was significantly higher than that in the other groups(P<0.0001,PBS vs.h UC-MSCs,De-Dif.P=0.002,h UC-MSCs vs.De-Dif),suggesting that De-Dif significantly improved the learning ability of HIBD rats compared with h UC-MSCs.(4)Through the Object-in-place experiment,the h UC-MSCs,Dif,De-Dif,Re-Dif transplantation group has significantly improved the ability to explore new things than the PBS group,and the difference is statistically significant(P = 0.002,PBS vs.h UC-MSCs,Dif,Re-Dif;P=0.000,PBS vs.De-Dif),and De-Dif has more obvious exploration ability(P=0.0017,De-Dif vs.h UC-MSCs;P=0.005,De-Dif vs.Dif;P=0.025,De-Dif vs.Re-Dif),indicating that the transplantation of De-Dif cells has stronger nerve repair ability in HIBD rats;(5)The whole gene GO analysis and KEGG pathway analysis were performed on h UC-MSCs and De-Dif.The results showed that after dedifferentiation induction of h UC-MSCs,the differentially expressed genes were mainly involved in the hypoxia regulation process,and a variety of cytokines including receptor serine/threonine kinases and CXCR chemokines were involved.(6)Based on the analysis of the two groups of differential genes,it was found that CXCL12 was the gene with the greatest difference between h UC-MSCs and De-Dif.RT-PCR verified this result,suggesting that the process of dedifferentiation mediated differential expression of SDF-1α,which may be involved in the biological function of De-Dif.(7)The RARs gene of the two groups was analyzed.Compared with h UC-MSCs,the expression of RARα was lower in the De-Dif group(P=0.030),and the expression of RARβ was significantly higher in the h UC-MSCs group(P=0.000).The difference was statistically significant.the RARβ level of the two groups of cells was verified by Western Blot,and it was found that the level of RARβ protein in the De-Dif group was significantly higher than that in the h UC-MSCs group(P=0.000),and the difference was statistically significant.It is suggested that RARβ may participate in the biological function of De-Dif;(8)According to the previous stage,the cell lines of h SDF-1α,both of which were underexpressed(sih SDF-1)and overexpressed(adh SDF-1),were successfully established.Western Blot was used to detect the expression of RARβ,and it was found that there was no statistical difference in the expression of RARβ between the two groups.Conclusion:(1)h UC-MSCs can successfully undergo neural differentiation,dedifferentiation and redifferentiation induction;dedifferentiated h UC-MSCs have stronger effect on HIBD nerve repair;(2)Dedifferentiated h UC-MSCs reprogram differentially express SDF-1α,and RARs genes,which may play a role in regulating the functional mechanism of HIBD nerve repair. |
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