Objective Type A lamins are encoded by LMNA and a major component of the nuclear lamina, which have been suggested to play important roles in chromatin organization, transcription, DNA replication, and cell apoptosis. The aim of this study was to analyze the bioinformation of zebrafish lamins. Methods A phylogeny analysis was figured out with protein sequences of different species by Clustal X and MEGA 4?0 software. Then we compared the lamin protein sequences of different species with that of zebrafish by BLAST tool from NCBI. A figure of synteny analysis results was done with lamin sequence information of humans, murine and zebrafish cited from UCSC, Vega and Ensemble. Results The a?nalysis results showed that lmna, lmnb1, and lmnb2 genes of zebrafish are highly conservative and they may be homology of human LMNA, LMNB1 and LMNB2 genes. Conclusions Zebrafish lamins and human lamins have homologous sequence similarity, indicating that these two genes are orthologous genes.

BACKGROUND:Polyhydroxybutyrate-co-volerate (PHBV) is a noticeable tissue engineering material of polyhydroxyalkanoates family. It has the properties of low immune rejection response and good biocompatibility, and its degradation products are non-toxic.
OBJECTIVE:To investigate the biocompatibility of PHBV membrane material and human bone marrow mesenchymal stem cel s in vitro.
METHODS:Human bone marrow mesenchymal stem cel s at passage 3 were seeded upon PHBV membrane as experimental group and upon conventional culture plates as control group. Then we calculated the adherent cel number of two groups at 1, 2 and 4 hours and got the cel adherent rate. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide assay was used at days 2, 4, 6, 8 to observe the cel proliferation of two groups. Fluorimetric method with the fluorescent dye Hoechst 33258 was used to detect the DNA content of cel s at days 3, 6, 9 and 12 in both groups. After cel s were seeded upon PHBV membrane for 5 days, the cel growth upon the material was examined under a scanning electron microscope.
RESULTS AND CONCLUSION:When the cel s were cultured for 1 hour, the adherent rate in the experimental group was lower than that in the control group;but there were no significant differences between two groups at the other two periods. No difference was found in the cel proliferation and the DNA content between the two groups. Human bone marrow mesenchymal stem cel s seeded upon PHBV membrane for 5 days grew wel with spindle morphology and the intercel ular connections were tight and more extracel ular matrices were observed by scanning electron microscopy. Taken together, PHBV membrane material shows a good biocompatibility with human bone marrow mesenchymal stem cel s.

Objective To explore the ability for constructing tissue engineering bone in vivo in complex scaffolds with PHB‐HHx as the scaffolds material and human umbilical cord mensenchymal stem cells (hUCMCs) as the seed cells .Methods hUCM‐SCs were inoculated into PHBHHx scaffolds to induce osteogenesis culture in vivo for two weeks ,the the induced group was the experimental group and those without instilling hUCMCs served as the control group ,the nude mouse subcutaneous implantation was performed .Then taking material at 1 ,3 ,5 months after implantation in vivo was performed for conducting HE ,collagenⅠim‐munohistochemical ,alkaline phosphatase staining and RT‐PCR .Results hUCMSCs showed good cellular adsorbability .The size and form in the experimental group basically maintained the original status ,and the osteogenesis specific indicators were positive ;but the control group did not keek the original status ,its volume was gradually shrunk until complete degradation ,and the osteogen‐esis specific indicators were negative .Conclusion The PHBHHx scaffolds combined with hUCMSCs has the capability of in vivo heterotopic constructing tissue engineering bone in nude mouse by in vitro osteogenic induction .

BACKGROUND:As a noticeable tissue engineering material of polyhy droxyalka noates family, poly (3-hydroxybutyrate-co-4-hydroxybutyrate)(P3HB4HB) exhibitsgood biocompatibility, adhesion and mechanicalproperties, presenting aextensive application future in tissue-engineered research.
OBJECTIVE:To investigate the biocompat ibilityin vitroand ectopic osteogenic differentiationin vivoof P3HB4HB and human bone marrow mesenchymal stem cels.
METHODS:Passage 5human bone marrow mesenchymal stem cels transplanted ontothe three-dimensional P3HB4HB scaffoldwereincubated with osteogenic induction medium (test group)or with no osteogenic induction(control group), respectively. After 5-day incubation, thecelgrowth was assessed by acridine orange staining and scanning electron microscopy; after14-day incubation, both kinds of cel-scaffold composites were subcutaneously implanted into the nude mice. At 16 weeks after implantation, the cel-scaffold composites were removed to observeectopic osteogenic differentiationin vivousing hematoxylin-eosin staining, von Kossa staining and colagen type I immunohistochemical staining.
RESULTS AND CONCLUSION:Acridine orange staining showed that cels adhered wel on the surface of the scaffold;under thescanning electron microscope, induced celsgrew wel on the P3HB4HB scaffold and produced abundant extracelular matrixes. In addition, at 16 weeks after implantation, there were osteoidtissues in the test group, positive for von Kossa staining as wel as colagen type I immunohistochemical staining;furthermore, hematoxylin-eosin staining showednumerous osteoblasts and bone lacunas. In contrast, no bone tissues appeared in the control group. To conclude, P3HB4HB is a suitable material for bone tissue engineering.

Objective To investigate the value of interleukin-6(IL-6),procalcitonin(PCT)and immuno-globulin G(IgG)in evaluating the susceptibility and recurrence of bacterial pneumonia in children with hor-mone-sensitive primary nephrotic syndrome(PNS).Methods A total of 150 children with hormone-sensitive PNS admitted to the hospital from January 2019 to March 2023 were selected as the study objects.The pa-tients were divided into concurrent group and non-concurrent group according to whether bacterial pneumonia was complicated or not within 3 months.The baseline IL-6,PCT and IgG levels of the two groups were com-pared,and the factors affecting susceptibility to bacterial pneumonia were analyzed by Logistic regression.Re-ceiver operating characteristic curve(ROC)was drawn to analyze the value of IL-6,PCT and IgG in assessing susceptibility to PNS bacterial pneumonia and the risk of PNS recurrence.Results IL-6[(7.84±2.49)pg/mL]and PCT[(0.39±0.11)ng/mL]in concurrent group were higher than those in non-concurrent group[(4.75±1.56)pg/mL,(0.29±0.09)ng/mL].IgG[(3.82±1.25)g/L]was lower than that of non-concurrent group[(5.92±1.97)g/L],and the differences were statistically significant(P＜0.05).Logistic regression e-quation showed that with the increase of IL-6 and PCT and the decrease of IgG,the susceptibility to bacterial pneumonia gradually increased,and the difference was statistically significant(P＜0.05).The AUC of IL-6,PCT and IgG for PNS bacterial pneumonia susceptibility was 0.792,0.744 and 0.677,respectively.The AUC of IL-6 and PCT combined IgG for PNS bacterial pneumonia susceptibility was 0.929,which was higher than that of IL-6,PCT and IgG alone(P＜0.05).The risk of PNS recurrence in patients with high IL-6 level was 7.054 times that of low IL-6 level,the risk of PNS recurrence in patients with high PCT level was 6.750 times that of low PCT level,and the risk of PNS recurrence in patients with high IgG level was 0.079 times that of low level,the differences were statistically significant(P＜0.05).Conclusion IL-6,PCT and IgG are the influencing factors of hormone-sensitive PNS bacterial pneumonia in children,and they are related to the recurrence of PNS.Clinically,IL-6,PCT and IgG levels can be used to evaluate the susceptibility and recur-rence risk of bacterial pneumonia in children,so as to provide guidance for preventive clinical intervention.

Objective To investigate the application effect of case-based teaching versus traditional teaching in the optional course of maternal and child nutrition for medical undergraduates.Methods The students who completed the course through the traditional teaching method in 2016 were enrolled as control group,and those who completed the course through the case-based teaching method in 2017 were enrolled as experimental group.In the process of teaching,the teachers in the experimental group needed to write cases,prepare lessons collectively,give guidance and inspiration,and summarize after class.During the learning process,the students in the experimental group needed to collect information before class and perform role play,group discussion,and intergroup communication about the case during class.After the course ended,a questionnaire for nutrition during pregnancy preparation,pregnancy,and lactation,breastfeeding,and complementary feeding was developed for assessment.With sex and grade as related factors,SPSS 18.0 was used for the chi-square test of two sample rates according to knowledge points,and the correct rate of each item was analyzed.Results A total of 107 valid questionnaires were collected from the control group,and 109 valid questionnaires were collected from the experimental group.For the questions on the knowledge of maternal and child nutrition,the experimental group had slightly higher correct rates than the control group (all P＞0.05).For the questions on the comprehension and practical application of the knowledge of maternal and child nutrition,the experimental group had significantly higher correct rates than the control group (all P＜0.05).Further stratified analysis based on sex and grade showed that the experimental group had higher correct rates than the control group.Conclusion Compared with traditional teaching,case-based teaching may not have advantages in enhancing students' memorizing ability,but it can make it easier for students to understand the theoretical knowledge of matemal and child nutrition and master the practical application of such knowledge.

Objective To investigate the influence of maternal high-protein diet during pregnancy and lactation on the growth of male rat offspring.Methods Female Wistar rats were mated with male rats and the pregnant ones were randomly assigned into three groups (n=10):Group Ⅰ,Group Ⅱ and Group Ⅲ.Three isocaloric purified diets,which were Diet Ⅰ (protein,14.0％;carbohydrate,69.3％;fat,16.7％),Diet Ⅱ (protein,24.0％;carbohydrate,59.3％;fat,16.7％) and Diet Ⅲ (protein,34.0％;carbohydrate,49.3％;fat,16.7％),were offered ad libitum to the rats in the three groups,respectively.On the 3rd day after birth,only three male rat offspring of each litter were kept.All male rat offspring at the age of 21 days were weaned onto the same normal diets until 77-day-old.Food intake (FI),body weight (BW),body length (BL) and visceral fat mass (VFM) of them were recorded.Blood lipids levels on the 21st,49th and 77th days after birth were detected.Weight gain (WG),food efficiency (FE) and percentage of VFM (VFM ％) were calculated to assess the growth of male rat offspring.One-way ANOVA,LSD or Tamhane T2 test was used for statistical analysis.Results (1) On the 3rd day after birth,male rat offspring in Group Ⅱ and Group Ⅲ had higher fast BW than those in Group Ⅰ [(9.77±0.81),(9.58±0.54) and (9.17±0.68) g;F=3.222,P=0.026].On the 7th,14th and 21st days after birth,male rat offspring in Group Ⅱ [(17.59± 1.24),(39.51 ±2.68) and (67.77±4.22) g] had higher BW than those in Group Ⅰ [(15.96±1.17),(35.35±4.11) and (63.43±3.98) g] and Group Ⅲ [(16.52±1.05),(37.06±3.27) and (64.43±3.81) g] (F=23.684,17.070 and 84.195,all P＜0.001).Male rat offspring's BL of Group Ⅱ were longer than those of Group Ⅰ on the 7th and 14th days after birth[(7.53±0.29) vs (7.53± 0.29) cm,(10.38 ±0.24) vs (9.99± 0.30) em,both P＜0.05].BL of Group Ⅲ was longer than that of Group Ⅰ,but shorter than that of Group Ⅲ on the 14th day after birth[(10.22 ± 0.25) vs (9.99± 0.30) cm,(10.22 ± 0.25) vs (10.38±0.24) cm,both P＜0.05].Male rat offspring's average WG from the 3rd to the 7th and the 7th to the 14th day after birth were higher in Group Ⅱ [(7.70±0.41) and (22.08±1.20) g] and Group Ⅲ [(7.00±0.40) and (20.75± 1.72) g] than in Group Ⅰ [(6.73±0.55) and (19.68± 1.73) g] (F=86.925 and 38.876,both P＜0.001),and the differences between Group Ⅱ and Group Ⅲ were statistically significant (both P＜0.05).(2) No significant differences in WG,FI and FE was observed among the three groups (all P＞0.05).Male rat offspring's BL on the 49th day after birth was longer in Group Ⅱ than in Group Ⅰ and Group Ⅲ [(22.03±0.26),(21.57±0.43) and (21.77±0.33) cm,F=3.222,P=0.026).VFM ％ of Group Ⅱ (3.87±0.32 and 5.13±0.32) and Group Ⅲ (3.90±0.27 and 5.15±0.33) on the 49th and 77th days after birth were higher than those of Group Ⅰ (3.50±0.34 and 4.68±0.38) (F=3.631 and 3.611,both P＜0.05).(3) Triglyceride (TG) level was higher in Group Ⅰ than in Group Ⅱ and Group Ⅲ on the 21st day after birth [(1.12±0.13),(0.89±0.10) and (0.97±0.12) mmol/L,F=7.283,P=0.004].However,Group Ⅱ and Group Ⅲ had a higher level of TG than Group Ⅰ on the 77th day after birth[(2.64±0.37),(2.43±0.32) and (1.90±0.21) mmol/L,F=12.321,P＜0.001].Conclusion Maternal high-protein diet can increase the birth weight of male rat offspring to a certain extent,which is influenced by carbohydrate content.Moreover,male rat offspring of dams fed with high-protein diet during pregnancy and lactation will have increased visceral fat accumulation and serum TG level during adulthood.

BACKGROUND:Spleen has the functions of blood storage,hematopoiesis,and immunity.With the increase of age,the structural degeneration and functional decline of spleen lead to the impairment of immune system function,thus accelerating the aging process of the body.The treatment of spleen aging in tree shrews with highly active umbilical cord mesenchymal stem cells has not been reported. OBJECTIVE:To explore the intervention effect and mechanism of highly active umbilical cord mesenchymal stem cells on spleen aging in tree shrews. METHODS:Highly active umbilical cord mesenchymal stem cells were isolated,cultured,and obtained from the umbilical cord tissue of newborn tree shrews by caesarean section.The differentiation abilities of adipogenesis,osteogenesis,and chondrogenesis were detected by three-line differentiation kit.Cell cycle and surface markers were detected by flow cytometry.The second generation of highly active umbilical cord mesenchymal stem cells were transfected with Genechem Green Fluorescent Protein with infection complex values of 100,120,140,160,180,and 200,respectively,to screen the best transfection conditions.After transfection,the fourth generation of highly active umbilical cord mesenchymal stem cells was injected into the tail vein of tree shrews in the elderly treatment group.The young control group and the aged model group were not given special treatment.After 4 months of treatment,the spleen tissue was taken and the structure of the spleen was observed by hematoxylin-eosin staining.β-Galactosidase staining was used to detect the activity of aging-related galactosidase.Immunohistochemical staining was used to detect the expression levels of p21 and p53 proteins.Ki67 and PCNA immunofluorescence staining was used to detect cell proliferation activity.Immunofluorescence staining was used to detect the expression levels of spleen autophagy protein molecules Beclin 1 and APG5L/ATG5.Reactive oxygen species fluorescence staining was used to detect the content of reactive oxygen species in spleen tissue.CD3 immunofluorescence staining was used to detect the change of the proportion of total T lymphocytes.The secretion levels of interleukin 1β and transforming growth factor β1 in spleen were detected by enzyme linked immunosorbent assay.The distribution of highly active umbilical cord mesenchymal stem cells labeled with green fluorescent protein in spleen tissue was observed by DAPI double staining of nucleus. RESULTS AND CONCLUSION:(1)Highly active umbilical cord mesenchymal stem cells grew in a short spindle shape with fish-like growth,with a large proportion of G0/G1 phase,and had the potential to differentiate into adipogenesis,osteogenesis,and chondrogenesis.(2)Multiplicity of infection=140 and transfection for 72 hours were the best conditions for labeling tree shrews highly active umbilical cord mesenchymal stem cells with Genechem Green Fluorescent Protein.(3)Compared with the aged model group,in the aged treatment group,the spleen tissue cells of tree shrews were arranged closely,and the area of white pulp was increased(P<0.01);the boundary between red pulp and white pulp was clear;the proportion of germinal centers did not show statistically significant difference(P>0.05).The activity level of galactosidase related to spleen tissue aging was decreased(P<0.001),and the expression levels of aging protein molecules p21 and p53 were down-regulated(P<0.001).The expression levels of proliferation-related molecules Ki67 and PCNA were up-regulated(P<0.001,P<0.05);expression levels of autophagy-related molecules Beclin 1 and APG5L/ATG5 were up-regulated(P<0.001),and the content of reactive oxygen species decreased(P<0.001),and the proportion of CD3+T cells increased(P<0.05).The secretion level of interleukin 1β in the aging-related secretion phenotype decreased(P<0.001);no significant difference was found in transforming growth factor β1 level(P>0.05).Compared with the young control group,the above indexes were significantly different in the elderly treatment group(P<0.05).(4)Green fluorescent cells labeled with green fluorescent protein were observed in spleen tissue of tree shrews the elderly treatment group by frozen tissue section observation.The results show that intravenous infusion of highly active umbilical cord mesenchymal stem cells can migrate to spleen tissue,inhibit the production of reactive oxygen species,down-regulate the expression of aging-related proteins,induce autophagy,promote cell proliferation,reduce chronic inflammation,and then improve the structure and function of spleen tissue.