Objective To investigate the results of graft repair of peripheral nerve defects using small intestinal submucosa(SIS) with crude Schwann cells. Methods Thirty-six male SD rats were randomly divided into three groups,with 12 in each group.Firstly,12-mm gaps of sciatic nerve were made in rats and then treated respectively with porcine SIS(group A),SIS compounded with crude Schwann cells(group B) and auto-nerve grafting(group C) in the three groups.The grafts were harvested 16 weeks postoperatively to evaluate the results of nerve regeneration through histological examination, triceps surae weight,computerized imaging analysis,Trueblue retrograde tracing and transmission electron microscopy. Results Nerve regeneration was confirmed to have extended through the gaps according to the results of histological examination,Trueblue retrograde tracing and transmission electron microscopy.Furthermore,in the respect of triceps surae weight,quantity of axis cylinder per area and percentage of neural tissue,the results of group B and group C were superior to those of group A with a significantly statistical difference(P0.05). Conclusion SIS compounded crude Schwann cells was able to achieve the outcome of auto-nerve grafting and was a promising replacement graft.

Objective To study the characteristics of dynamic susceptibility-contrast(DSC)MR perfusion curves,color images and perfusion values in pre-operative brain metastasis.Methods Twenty- eight brain metastases underwent DSC MR perfusion imaging by using a first-pass T_2~* echo-planar sequence. The patients' data were transferred to on-line workstation.Time-signal intensity curves,color perfusion maps and rCBV,rMTT values in both tumor parenchyma and peri-tumor edema were analyzed,and independent t- test was used and P0.05).Conclusion Different originated brain metastases have nearly same characteristics in DSC MR perfusion imaging.

Objective To observe adhesion and growth of Sehwann cells(SCs) on small intestinal submueosa(SIS) and study the bioeompatibility of SIS with SCs.Methods The SCs of SD neonate rat were isolated and cultured in vitro,then were seeded on prepared SIS.At different times,the adhesion,growth and proliferation of SCs on SIS were observed by phase contrast microscope,histological examination,scanning e- lectron microscope and transmission electron microscope.Results By phase contrast microscope,SCs grew well on the edge of SIS after 3 and 5 days.SCs adhered tightly on the surface of SIS after 5 days through histo- logical examination.By scanning electron microscope,on the surface of SIS,SCs grew and adhered actively, prominence of cells body were obvious.They connected end-to-end with each other or arranged in clusters. Protein granules were secreted on cells surface.By transmission electron microscope,SCs grew in good condi- tion and adhered tightly on the surface of SIS.At the interface of SCs and SIS,prominence was seen to contact with SIS in the bottom of cell body.Conclusion SCs are able to adhere and grow well on the surface of SIS.As a scaffold,SIS has good biocompatibility with SCs.

Cell cycle progression is tightly regulated in hematopoietic stem cells. The cycle state decides cells' fates, which includes self-renewal, proliferation and differentiation. Proper cell cycle regulation is a pivotal element for the maintenance of hematopoiesis homeostasis. HTm4 is a newly identified specific cell cycle regulator of the hematopoietic cell. Through interacting with KAP-CDK2 complex, it arrests cells in G(0)/G(1) phase. K562 is a human chronic myelogenous leukemia cell; it could be induced to megakaryoblast by phorbol 12-myristate 13-acetate (PMA). Such differentiation must be associated with cell cycle change. To further clarify HTm4's function in hematopoietic cell cycle regulation, K562 cells were treated with PMA. Cell cycle change was analysed using flow cytometric system. And during the induction process gene expression of HTm4 as well as CycleE and CDK2, which are responsible for G(1) to S transition, were analysed using semi-quantitative RT-PCR. The C-terminal domain of HTm4 protein has been shown to be important for HTm4's binding with KAP-CDK2 complex. To determine its impact on HTm4's function, HTm4 and C-terminal truncated HTm4 (HTm4-ct) were transfected into K562 cells using Tet-Off regulation expression system. Their influence on cell cycle was observed. The results showed that PMA induced both expansion and differentiation of K562 cells as measured by cell number count and NBT staining respectively. During PMA treatment, G(0)/G(1) cell proportion and HTm4 expression displayed coordinated change, which suggested that HTm4 might drive K562 cells out of cell cycle but was not involved in the quiescence maintenance. Additionally, transfection of HTm4 caused G(0)/G(1) arrest in K562 cells, while transfection of HTm4-ct did not. It is therefore suggested that the C-terminal domain is important for the function of HTm4 in cell cycle regulation.
Cell Cycle ; physiology ; Cell Cycle Proteins ; genetics ; physiology ; Cells, Cultured ; Gene Expression Regulation ; Hematopoiesis ; physiology ; Hematopoietic Stem Cells ; cytology ; physiology ; Humans ; K562 Cells ; Membrane Proteins ; genetics ; physiology ; Tetradecanoylphorbol Acetate ; pharmacology ; Transfection

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Female ; Hepatitis B virus ; physiology ; Hepatitis B, Chronic ; virology ; Humans ; Male ; Middle Aged ; Virus Replication

Objective To investigate the effects of physical exercises on cardiac function and plasma N-terminal pro-brain natriuretic peptide (NT-proBNP) levels in hypertensive patients combined with diastolic cardiac dysfunction.MethodsA total of 66 essential hypertension patients who had abnormal left ventricular relaxation and normal systolic function were assigned to the intervention group ( n =33 ; doing physical exercises once a day,5 days a week) or control group (n =33 ).All the patients received standard treatment.At 6 months,body weight,blood pressure,heart rate,NT-proBNP,and echocardiography were measured.ResultsAt 6 months,body weight [ (68 ± 7 ) kg vs (72 ± 8 ) kg ],systolic blood pressure [ (135.4 ±5.1) mm Hg (1 mm Hg =0.133 kPa) vs (141.9 ±5.2) mm Hg ],diastolic blood pressure [ (81.1 ±4.0) mm Hg vs (84.7 ±4.6) mm Hg],New York Heart Association class (1.4 ±0.3 vs 1.8 ±0.4),NT-proBNP level [ (526 ± 126 ) ng/L vs (741 ± 189 ) ng/L] were significantly decreased in the intervention group when compared with the control group ( all P ＜ 0.05 ) although left ventricular ejection fraction (LVEF) (62.9 ±6.7 vs 59.0 ±5.6) and E/A ratio ( 1.1 ±0.3 vs 0.9 ±0.3) were significantly increased ( both P ＜ 0.05).ConclusionPhysical exercises could play a role in reduced blood pressure and body weight and improved cardiac function in hypertensive patients with diastolic cardiac dysfunction.

OBJECTIVE: To explore the characteristics of Shwachman-Diamond syndrome (SDS) in Chinese children in order to provide a reference for early diagnosis. METHODS: With Shwachman-Diamond syndrome, SDS, SBDS gene and inherited bone marrow failure as the keywords, the search period was set from January 2002 to October 2022. Relevant literature was retrieved from the Wanfang Database and China National Knowledge Infrastructure (CNKI) database. In addition, by using Shwachman-diamond syndrome as a keyword, the search period was also retrieved from the Web of Science, PubMed, and MEDLINE databases from January 2002 to October 2022. A child with SDS treated at the Tongji Hospital was also included. A total of 44 cases with complete clinical data were analyzed with reference to the International Standard for SDS Diagnosis. Chi-square test and t test were used for statistical analysis. Evidence-based research was carried out in the form of systematic review. The epidemiology, clinical characteristics and key points of early diagnosis of the Chinese SDS children were summarized and compared with the international data. RESULTS: The main characteristics of SDS in Chinese children were summarized as follows: The ratio of males to females was about 1.3 : 1, the median age of onset was 3 months, and the median age of diagnosis was 14 months. The first symptoms were often exocrine pancreatic insufficiency (31.8%) and granulocytopenia with infection (31.8%). According to the international consensus, the incidence rates of the three major diseases of SDS were hemocytopenia (95.4%), pancreatic disease (72.7%), and bone abnormality (40.9%). The common factors underlying SDS disease were variants of the SBDS gene (c.258+2T>C and c.183_184TA>CT), albeit there was no significant correlation between genotype and phenotype (P > 0.05). Compared with international reports, the clinical manifestations and genotypes of Chinese SDS children are different (P < 0.05). CONCLUSION The SDS children have an early age of onset and significant individual difference. It is necessary to analyze the case-related data to facilitate early recognition, diagnosis and clinical intervention.
Female ; Humans ; Male ; Bone Marrow Diseases/therapy* ; China ; East Asian People ; Exocrine Pancreatic Insufficiency/therapy* ; Shwachman-Diamond Syndrome/therapy*

E2 gene of classical swine fever virus (CSFV) was cloned into secretory pPIC9K Pichia pastoris expression vector. After being linearized by digestion, the vector was transformed into Pichia pastoris by electroporation to integrate with the genome, the transformants with high copies were screened by G418 and were induced to express with methonal. The results of SDS-PAGE and Western blot demonstrated that the supernatant of the induced P. pastoris culture contained protein E2. The results of the study on the immunological activity indicated that the protein E2 expressed in P. pastoris can elicit animal bodies to produce antibodies against protein E2.
Animals ; Antibodies, Viral ; immunology ; Antigens, Viral ; genetics ; immunology ; Classical swine fever virus ; genetics ; immunology ; Cloning, Molecular ; Gene Expression ; Pichia ; Rabbits ; Swine ; Viral Envelope Proteins ; genetics ; immunology

The major antigen region of E2 gene of Hog Cholera Prevalent Strain (Guangxi Yuling Strain) and Chinese Hog Cholera Lapinised Virus (C-strain) derived from hog and rabbit spleen tissue, was amplified by reverse transcription polymerase chain reaction(RT-PCR) and the nested Polymerase Chain Reaction (nPCR). After the amplified fragments were cloned into the expression vector pPROEX-HTb, the recombinant plasmids pPROEX-GXYL and pPROEX-C were obtained. The insert position, the size and the reading frame were right by PCR, restriction digestion and the sequence analysis. SDS-PAGE indicated that both of the reciepient germs transducted and induced by the recombinant plasmids pPROEX-GXYL and pPROEX-C could express the major antigen region of E2 gene. Western-blot indicated that the expressed antigen protein could be recognized by the positive serum of CSFV.
Blotting, Western ; Cloning, Molecular ; Escherichia coli ; genetics ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; Viral Envelope Proteins ; biosynthesis ; genetics ; immunology

OBJECTIVETo explore the mechanism of Flt3 receptor-interacting lectin (FRIL) maintains quiescence of hematopoietic stem cells (HSCs) in vitro.

METHODSCord blood CD34+ cells were cultured in suspension medium supplemented with or without FRIL and FL. Cells were collected at different time points and the expression of some cell cycle regulators, especially those involved in G0/G1 phase regulation were detected on mRNA and protein level.

RESULTSThe expressions of G0/G1 phase related cyclins or CDKs were undetectable in the newly isolated CD34+ cells, expressions of Cyclin D3, CDK6 and P27 were the lowest in FRIL cultured group after 3d's culture (FRIL group: 483 +/- 63, 553 +/- 39, 0.312 +/- 0.030; FL group: 2437 +/- 52, 3209 +/- 98, 0.787 +/- 0.024; BLANK: 914 +/- 105, 1497 +/- 55, 0.616 +/- 0.029, respectively), but the expression of P53 was the highest in FRIL group (FRIL group: 4.476 +/- 0.159; FL group: 0.581 +/- 0.099, BLANK: 2.167 +/- 0.114). The expression of positive regulators of cell cycle in FRIL group were the same as that of FL group and blank group or lower.

CONCLUSIONFRIL preserves HSCs effectively in vitro through the mechanisms of down-regulation of cyclin D3 and CDK6 and activation of P53. P27 is mostly involved in the differentiation of HSCs.

Antigens, CD34 ; Cell Cycle ; drug effects ; Cell Cycle Proteins ; genetics ; metabolism ; Cells, Cultured ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; cytology ; drug effects ; metabolism ; Humans ; Mannose-Binding Lectins ; pharmacology ; Plant Lectins ; pharmacology ; RNA, Messenger ; genetics