BACKGROUND:The metastatic potential of hepatocelular carcinoma cels is key factor influencing patient’s prognosis. To observe the effect of human bone marrow mesenchymal stem cels on metastasis of hepatocelular carcinoma is of great significance for improving the lifetime of hepatocelular carcinoma patients. OBJECTIVE:To explore the biological effect of human bone marrow mesenchymal stem cels on hepatocelular carcinoma cels with different metastatic potentials. METHODS:Human bone marrow mesenchymal stem cels and hepatocelular carcinoma cel suspension with high and low metastatic potentials were respectively injected into the Transwel chamber, and after 36 hours of co-culture, ELISA method was used to detect the absorbance value as wel as cel counting method was used to observe the changes in the invasion ability of hepatocelular carcinoma cels. The effects of human bone marrow mesenchymal stem cels on the proliferation of hepatocelular carcinoma cel suspension with high and low metastatic potentials were determined using cel counting kit-8. PCR method was adopted to measure the expression of osteopontin, bone specific sialoproteins, integration (alpha V), transforming growth factor beta 1 and programmed cel death protein 5. RESULTS AND CONCLUSION:(1) The number of migrated hepatocelular carcinoma cels was significantly lower in the co-culture group than the single culture group, and based on the semi-quantitative detection of invasion ability, the absorbance value of the co-culture group was significantly lower than that in the single culture group (P < 0.05). (2) The expression of osteopontin and bone specific sialoproteins was significantly decreased in the co-culture group with high metastatic potential (P < 0.05), but there was no change in the expression of integration (alpha V) (P> 0.05). In the co-culture group with low metastatic potential, the expression of osteopontin, bone specific sialoproteins, and integration (alpha V) were declined remarkably (P < 0.05). (3) Results from the semi-quantitative detection of proliferation ability showed that the absorbance value of the co-culture group was significantly higher than that of the single culture group (P < 0.05). (4) In the co-culture group with high metastatic potential, the expression of transforming growth factor beta 1 was up-regulated significantly (P< 0.05), but the expression of programmed cel death protein 5 showed no changes (P > 0.05). However, in the co-culture group with low metastatic potential, the expression of transforming growth factor beta 1 and programmed cel death protein 5 was both increased dramaticaly (P < 0.05). These findings suggest that the human bone marrow mesenchymal stem cels reduce the invasion ability of hepatocelular carcinoma cels, and enhance their ability of proliferation.

Objective To investigate the influence of in vitro artificial CO 2 cavity on matrix metallopro-teinase 2(MMP-2), adhesion molecule vascular cell adhesion molecule 1(VCAM-1), and intercellular adhesion molecule 1(ICAM-1)expression in MDA-MB-231 cell.Methods An in vitro artificial CO2 cavity model was es-tablished.MDA-MB-231 cells were exposed to CO2 under the pressure of 7 mmHg for 1, 2 and 4 hours, respective-ly.MMP-2 concentration was measured by enzyme linked immunosorbent assay (ELISA).VCAM-1 and ICAM-1 ex-pression were measured by flow cytometry 0, 24, 48 and 72 hours after CO2-insufflation.Hypoxia group was ex-posed to 0 mmHg helium for 1 h, and the control group was exposed to 37℃incubator only .Results Compared with that in the control group , MMP-2 expression in the 1, 2 and 4 hours treatment group was significantly elevat-ed at 0 hour(F=15.045, P<0.05), and the MMP-2 expression in the 2 hours CO2 treatment group was signifi-cantly elevated after 24 hours compared with that in the control group and 1, and 4 hours CO2 treatment group (F=5.976, P<0.05).The VCAM-1 expression was significantly elevated at 0 hour and after 24 hours in the 1, 2 and 4 hours CO2 treatment group compared with that in the control group ( F1 =18.321, F2 =20.443, P<0.05), and significantly declined after 72 hours in the 4 hours CO2 treatment group compared with that in 1, and 2 hours CO2 treatment group(F=15.045,P<0.05).ICAM-1 expression was significantly elevated at 0 hour in hypoxia group and 1, 2, 4 hours CO2 treatment group compared with that in the control group , Meanwhile it was higher in 2 hours CO2 treatment group than in 1 hours and 4 hours CO2 treatment group(F=73.765, P<0.05). ICAM-1 expression was significantly elevated after 24 hours in 2 and 4 hours CO2 treatment group compared with that in the control group and 1 hour CO2 treatment group(F=46.322, P<0.05), and it was significantly elevat-ed after 48 hours in 2 hours CO2 treatment group compared with that in the control group and 1, and 4 hours CO2 treatment group(F=22.315, P<0.05).Conclusion The expression of MMP-2, adhesion molecule VCAM-1, and ICAM-1 in MDA-MB-231 cells is elevated after exposure to artificial 7 mmHg CO2 cavity, and CO2 cavity of mastoscopy may modulate the metastasis capacity of breast tumor cells .

Objective To evaluate the expression of CD47 and calreticulin and their relationships with clini-copathologic features,prognosis in infiltrating ductal breast carcinomas.Methods The expression of CD47 and calret-iculin was evaluated in 128 cases with infiltrating ductal breast carcinomas by means of immunohistochemistry,and the relationship between CD47 and other clinicopathologic factors as well as prognosis were analyzed.Results The expression level of CD47 in cancer nest was 81/128,which was lower than 106/128 in its adjacent normal epithelium (χ2 =12.400,P<0.05),but calreticulin expression was 50/128 and was higher in cancer nest(χ2 =21.510,P<0.05 ).In addition,the expression level of calreticulin had certain difference in different molecular subtype (χ2 =21.510,P<0.05),and an increased expression in Basal-like and Her-(2 +)/ER(-)subtypes.The expression level of CD47 was associated with tumor sizes,histological tumor grade and lymph node involvement(χ2 =11.400, 4.732,5.432,all P<0.05),and the expression level of calreticulin was associated with histological tumor grade and distant metastasis(χ2 =10.810,6.770,all P<0.05).The 5 -year survival rate of patients performed following-up was 68.37%.Univariate analysis indicated the survival rate in 5 years of high level CD47,calreticulin expression was higher than that of low level CD47,calreticulin expression(χ2 =5.231,13.069,all P<0.05).Conclusion The expression of CD47,calreticulin is associated with index of dysprognosis and survival rate in 5 years,so combined CD47 and calreticulin detect may be of prognostic value in patients with breast caner.

In order to solve the problesm in biological sealing of load-bearing percutaneous implants for a fairly long time, we investigated titanium with bioactivated anodic oxidized surface(group A) through the animal tests in vivo and the epithelium cell culture in vitro. Smooth Ti (group B) was used as control. The animal tests results showed that there was no evident difference in the inflammory reaction between the group A implant tissues and the group B implant/tissues. The bioactivated Ti surface could keep the implant not only bonding with the bone firmly but also adhering to the soft tissue closely, thus contributing to the formation of calcium phosphate layer and its micropores. The cell culture results also demonstrated that the microporous surface of group A could clasp and fix the skin. So, it can be concluded that the surface modified method of anode oxidization may be one of the most effective methods to resolve the problem of durable biological sealing.
Animals ; Biocompatible Materials ; chemistry ; Cells, Cultured ; Coated Materials, Biocompatible ; Epithelial Cells ; drug effects ; Materials Testing ; Osseointegration ; Prostheses and Implants ; Tibia ; surgery ; Titanium ; chemistry

Objective To construct a collagen matrix-based skin model that can last a long time in vitro for potential application of subsequent tests and studies.Methods The porcine collagen and normal human skin fibroblasts were mixed and seeded into the culture dish,which were cultured for 3-4 days to obtain a dermal structure.Then,the third-to seventh-passage normal human keratinocytes were seeded onto the dermal surface and cultured for 14 days to obtain a double-layer skin model.Hematoxylin and eosin (HE)staining and Masson staining were performed to evaluate the morphology and structure of the skin model,and electron microscopy was conducted to observe the ultrastructure of the skin model.Immunohistochemical study and immunofluorescence staining were conducted to determine the expression of major markers in each layer of the skin model.Results HE and Masson staining showed that the skin model and normal human skin tissues showed very similar dermal and epidermal structures.After harvest of the skin model,it can be cultured in vitro for another 14 days with favorable and stable dermal and epidermal structures.Electron microscopy showed lipids in the stratum corneum,keratohyalin granules in the stratum granulosum,corneodesmosomes,desmosomes and basal membrane in the skin model.Immunohistochemical and immunofluorescence staining showed the consistent expression of transglutaminase,filaggrin,keratin 10 and Ki67 in the epidermis in both the skin model and normal skin.Moreover,the expression of type Ⅳ collagen and laminin-5 in the basal membrane,as well as that of type Ⅰ collagen,type Ⅲ collagen and fibrillin in the dermis were both consistent between the skin model and normal skin.Conclusion The constructed three-dimensional (3D) skin model is highly analogous to normal human skin in the aspect of tissue structure and various protein expression,and the skin model can be stably cultured in vitro for at least 14 days after harvest.

Objective To investigate changes in coagulation function and inflammation levels during sepsis.Methods A rat model of sepsis was established using the multiple infection sepsis model(MIM)based on cecal ligation and puncture.Forty-eight male Sprague-Dawley rats were assigned randomly to the following groups:control group,sham group,4 h sepsis group,8 h sepsis group,12 h sepsis group,and 16 h sepsis group(n=8 per group).Inflammatory markers and coagulation-related indicators were measured by enzyme-linked immunosorbent assay and coagulation analysis.Results(1)Lipopolysaccharide(LPS)and interleukin-6(IL-6)levels were significantly higher in the model rats at all time points compared with the sham group(P＜0.001).LPS and IL-6 levels increased gradually with disease progression,with no further changes in LPS after 12 hours.(2)Prothrombin time(PT)was significantly prolonged in the middle and late stages of the sepsis model,starting from 8,compared with the sham group(P＜0.01).(3)Partially activated prothrombin time(APTT)time was significantly prolonged in the 8,12,and 16 h groups compared with the sham group(P＜0.05,P＜0.01).APTT gradually lengthened from 8 h,but approached control levels thereafter.(4)Fibrinogen(Fbg)content was significantly higher in all sepsis groups,except for the 8 h group,compared with the sham group(P＜0.01).(5)Fibrin degradation products(FDP)differed significantly between the control and sham groups(P＜0.01),but not between the sham and sepsis groups.(6)Antithrombin-Ⅲ(AT-Ⅲ)levels decreased significantly throughout each stage of sepsis progression compared with the sham group(P＜0.01),and AT-Ⅲ showed a downward trend with the course of disease,with significant differences among the 4,8,and 16 h groups.Conclusions The MIM rat model can reflect the development of inflammatory and blood coagulation disorders and their relationship during the course of sepsis,and may thus provide a good foundation for further research into the disease course of sepsis.

Objective To investigate the epidemiological characteristics of cerebral palsy(CP)in children aged 1-6 years in China,including the incidence,prevalence,type of CP,etiology,prevention and rehabilitation status. Methods The survey was carried out by standard questionnaires,multi-center collaboration,stratified-cluster ran-dom sampling method.The surveyed adopted the following principles:streets in the city and villages in the rural areas, and the number of the urban and rural children was the same,and the proportion of children in each age group was balanced.The investigation areas included provinces and autonomous regions,including Heilongjiang,Beijing,Henan, Shandong,Shanxi,Shaanxi,Anhui,Hunan,Guangxi,Guangdong,Chongqing and Qinghai,and 323 858 children were in-vestigated.Results The incidence of CP was 2.48‰(155/62 591 cases),and the prevalence was 2.46‰(797/323 858 cases)(1-6 years old).The prevalence varied in different regions,in which the highest prevalence was 5. 40‰(54/9 998 cases)in Qinghai province,and the lowest prevalence was 1.04‰(47/45 133 cases)in Shandong province.The prevalence of the males(2.64‰,461/174 391 cases)was higher than that of the females(2.25‰, 336/149 467 cases),and the difference was statistically significant(P<0.05).The types of CP were spastic type (58.85%,469/797 cases),mixed type(13.17%,105/797 cases),dyskinetic(9.79%,78/797 cases),hypotonic (8.28%,66/797 cases),ataxia(6.25%,52/797 cases)and rigid(3.39%,27/797 cases)respectively in 797 CP children.The first three risk factors for CP were long -term exposure to harmful physical factors during pregnancy, whether there were birth defects among the three generations of relatives of the children,such as children's peers, parents or grandparents,whether there were birth defects among the children's peers,parents or grandparents,and neonatal jaundice or persistent jaundice.Among 797 CP children,79.67% of the children with CP were timely detected and treated in the local hospitals,while the other 19.93% of them were not timely treated.The places which could give them timely detection and early diagnosis and treatment were general hospitals(42.97%),Maternity and Infant Hospitals (27.03%)and Children's Hospitals(20.31%). The main rehabilitation methods for 797 children with CP were 34.58% in the hospitals or rehabilitation centers,31.61% in the communities(including at home),33.80% mainly in the medical institution,and in the communities they could also receive partially rehabilitation services. Conclusions The prevalence of CP in China is coincident with international levels.The prevalence rate of CP in males is higher than that in females.The types of CP distribution are accorded with international distribution characteristics.There were still some children with CP who could not receive timely detection and treatment.Rehabilitation at the medical institutions is the chief way and proper rehabilitation guidance should be carried out in the communities.

Currently, as the key raw material of artificial biological heart valve, bovine pericardium is mainly depend on import and has become a "bottleneck" challenge, greatly limiting the development of domestic biological heart valve. Therefore, the localization of bovine pericardium is extremely urgent. In this study, the pericardium of Sichuan yak was compared with that of Australian cattle in terms of fundamental properties and anti-calcification performance. The results demonstrated that the appearance and thickness of yak pericardium were more advantageous than the Australian one. Sichuan yak pericardium and Australian cattle pericardium had comparable performance in shrinkage temperature, mechanical test and anti-calcification test. This study preliminarily verifies the feasibility of substitution of Australian cattle pericardium by Sichuan yak pericardium and promotes the progression of bovine pericardium localization with data support.
Animals ; Australia ; Bioprosthesis ; Cattle ; Heart Valve Prosthesis ; Pericardium

Objective To evaluate the impact of optimizing the stroke green channel on the efficiency of acute ischemic stroke management in a county hospital. Methods A retrospective analysis of the emergency stroke green channel treatment data from Sixian People’s Hospital from May 2020 to April 2021 (before optimization of the green channel) and from May 2021 to April 2022 (after optimization of the green channel) was conducted. The rates of intravenous thrombolysis (IVT) and mechanical thrombectomy (MT) patients, as well as door-to-needle time (DNT), door-to-puncture time (DPT), and the modified Rankin scale (mRS) scores of patients three months post-treatment before and after the optimization of the stroke green channel were compared. Results Within one year before and after optimization of the green channel, the number of acute visits for ischemic stroke was 3 143 and 2 623, respectively. Before optimization, 84 and 51 underwent IVT and MT, respectively. After optimization of the green channel, the ratios of patients underwent IVT (n＝215) and MT (n＝103) significantly increased, and both DNT and DPT were significantly shortened (P＜0.000 1); the proportion of MT patients with an mRS score of 0-2 at 3 months post-discharge significantly increased (46/99 vs 13/46, P＝0.038). Conclusion After optimizing the green channel at Sixian People’s Hospital, the efficiency of stroke treatment has significantly improved, and the patients’ prognosis improved.

ObjectiveTo investigate the efficacy of Bushen Shengxue prescription and Yiqi Yangxue prescription in the treatment of chronic aplastic anemia and the effect on T cell subsets and the expression of T-box expressed in T cells （T-bet） and GATA binding protein 3 （GATA3）. MethodA total of 585 patients with chronic aplastic anemia who were treated in 19 hospitals in China from May 2018 to June 2021 were enrolled. With the prospective， double-blind and randomized control methods， the patients were randomized into three groups： kidney deficiency group， Qi and blood deficiency group， and control group. The three groups were respectively treated with Bushen Shengxue prescription granule， Yiqi Yangxue prescription granule， and Placebo （half the dose of Bushen Shengxue formula granules）. In addition， all of them were given oral cyclosporin and androgen. The treatment lasted 6 months， with 3 months as a course. The blood routine indexes， T cell subsets， and fusion genes T-bet and GATA3 before and after treatment were analyzed， and the safety indexes were monitored. ResultDuring the observation， a total of 75 cases dropped out and 18 were rejected. Finally， 161 cases in the kidney deficiency group， 164 in the Qi and blood deficiency group， and 167 in the control group were included. After 6 months of treatment， the total effective rate was 98.8% （159/161） in the kidney deficiency group， which was higher than the 79.9% （131/164） in the Qi and blood deficiency group （χ²=30.135， P<0.01） and the 61.7% （103/167） in the control group （χ²=70.126， P<0.01）. The total effective rate was higher in the Qi and blood deficiency group than in the control group （χ²=13.232， P<0.01）. After treatment， the hemoglobin （HGB） content increased significantly in three groups （P<0.05） as compared with that before treatment， particularly the kidney deficiency group （P<0.01）. After treatment， the white blood cell （WBC） count and platelet （PLT） count in the kidney deficiency group and the control group increased compared with those in the Qi and blood deficiency group （P<0.01）. There was no specific difference in neutrophils （ANC） after treatment among the three groups. At the same time point， the level of T helper type 1 （Th1） cells， Th1/Th2 ratio （P<0.05）， level of CD4⁺， and CD4⁺/CD8⁺ ratio （P<0.05） were significantly low in the kidney deficiency group among three groups. There was no significant difference in CD19^-， HLA/DR⁺， and CD25⁺ between the kidney deficiency group and the other two groups， but the T-bet of the kidney deficiency group and the control group was lower than that of the Qi and blood deficiency group （P<0.05）. ConclusionBushen Shengxue prescription exerts therapeutic effect on the aplastic anemia by improving the immunoregulatory mechanism， inhibiting the activity of immune system， modulating T cell subsets， suppressing Th1 and CD4⁺， and promoting bone marrow hematopoiesis. Moreover， it is safe with little side effects， which is worthy of further promotion.