Objective To investigate the effects of cryopreservation on the growth and osteogenesis capa- bility of human bone marrow stromal cells(BMSCs)on demineralized bene matrix(DBM).Methods Bone marrow aspirates were obtained from the lilac crests of three donors.The BMSCs were isolated from the bone marrow by density gradient centrifugation.Cells of passage 3 were cryopreserved in liquid nitrogen for 24 hours,and then re- covered.The non-cryopreserved BMSCs were used as the control,The cryopreserved and control BMSCs were cul- tured in osteogenic media,collected and labeled with Dil to be seeded onto the DBM when cells were confluent.The percentage of BMSCs adhered to the DMB was detected.The cell morphology and matrices secreted by BMSCs on the DBM were observed by the inverted phase-contrasted microscope,fluorescence microscope and scanning electron microscope(SEM).The growth and viability of BMSCs on the DBM were determined using the modified MTT ashy. The osteogenesis ability of BMSCs on the DBM was determined by assessment of the alkaline phosphatase(ALP) activity and osteocalcin(OCN)content.Results The percentages of the cryopreserved and control cells adhered to DBM were(97.25?1.17)% and(97.00?1.09)% respectively.The cells adhered well to the DBM and grew rapidly.Large amounts of matrices on the DBM were observed by the light microscope and SEM.The cells embedded in the matrices could be observed by fluorescence microscope.There were no significant differences in the assay values of MTT,ALP and OCN between the cryopreserved and control BMSCs on the DBM.Conclusion Since cryopreservation does not affect the growth and osteogenesis capability of BMSCs on DBM,the cryopreserved BMSCs can be used as a cell source in bone tissue engineering.

Objective: To comprehensively analyze the characteristics of posts released on WeChat official accounts of tertiary hospitals in Wuhan, and compare the two types of communication platforms varying in penetration power in terms of posts features and text contents. The aim is to identify post service strategies for enhancing influence of their WeChat accounts and to provide applied support for their operations regarding posts. Methods: A total of 2 668 full-text messages were collected from the WeChat accounts of 22 tertiary hospitals in Wuhan in 2017. The 22 hospitals were categorized into a high WCI group and an ordinary group via Pareto′s principle. Then content analysis and data statistical analysis among others were used to analyze and compare the contents of the two groups in such five dimensions as push time, push content, title feature, media type and originality based on the WCI index. Results: The push timing of WeChat articles focuses on the two time periods from 10: 00 to 12: 00 and from 16: 00 to 18: 00. These articles mostly aim at disseminating health knowledge (accounting for 37% of total articles) and describing hospital information (accounting for 29% of total articles). There were significant difference between the WeChat accounts of the two groups of hospitals in terms of title characteristics, media forms, and originality (P<0.01). Conclusions Hospitals can enhance their WeChat accounts′ penetration power and hospital influence by means of optimized push time, featured titles, enriched media forms, and customized articles.

To investigate the influence of bear bile on rat hepatocarcinoma induced by diethylnitrosamine (DEN), a total of 40 rats were randomly divided into 4 groups: normal control group, model group, and two bear bile treatment groups. The rat liver cancer model was induced by breeding with water containing 100 mg x L(-1) DEN for 14 weeks. The rats of the bear bile groups received bear bile powder (200 or 400 mg x kg(-1)) orally 5 times per week for 18 weeks. The general condition and the body weight of rats were examined every day. After 18 weeks the activities of serum alanine transaminase (ALT), aspartate transaminase (AST) and total bilirubin (TBIL) were detected. Meanwhile, the pathological changes of liver tissues were observed after H&E staining. The expression of proliferative cell nuclear antigen (PCNA) and a-smooth muscle actin (alpha-SMA) in liver tissue were detected by immunohistochemical method. After 4 weeks the body weights of rats in normal group were significantly more than that in other groups (P < 0.05); and that in the two bile groups was significantly more than that in the model group. Compared with normal group, the level of serum glutamic-pyruvic transaminase and total bilirubin increased significantly in other groups; compared with model group, these two indexes decreased significantly in two bile groups. Hepatocellular carcinoma occurred in all rats except for normal group; there were classic cirrhosis and cancer in model group while there were mild cirrhosis and high differentiation in two bile groups. There were almost no expressions of PCNA and alpha-SMA in normal group while there were high expressions in model group; the two bile groups had some expressions but were inferior to the model group, and alpha-SMA reduced markedly. It indicated that bear bile restrained the development of liver cancer during DEN inducing rat hepatocarcinoma, which may be related to its depressing hepatic stellate cell activation and relieving hepatic lesion and cirrhosis.

BACKGROUND: A lot of researches have proved that polyglicolide acid (PGA), as a stent material, has been successfully used to construct engineered tissues, such as cartilage, bone and tendons, in nude mice or even big mammal. Whether the incubation of vascular endothelial cells and smooth muscle cells in the polyglicolide acid may subcutaneously construct vessel-like structure in nude mice needs a further study.OBJECTIVE: To verify the feasibility of forming vessel-like structure in the nude mice by subcutaneous implantation of polyglicolide acid cocultured with vascular endothelial cells and smooth muscle cells derived from newborn umbilical vein.DESIGN: Contrast study.SETTING: Tissue Engineering Key Laboratory, Medical College, Shanghai Jiao Tong University.MATERIALS: This study was performed at Tissue Engineering Key Laboratory, Medical College, Shanghai Jiao Tong University from January to June 2002. Belly band was derived from newborn babies in our department of obstetrics & gynecology. The parturien provided the informed consent, and this study was approved by the local research ethics committee. Twenty-six nude mice (3-4 weeks old, clean grade, irrespective of gender) were selected in this study. The animal experiment received confirmed consent from the local ethic committee. Polyglicolide acid was provided by Albany International Research Co.METHODS: Vascular endothelial cells and smooth muscle cells derived from newborn umbilical vein were incubated on a piece of polyglicolide acid to produce cell-material compound. In addition, the compound covered around the silicone tube to form a tube-like structural substance. Subsequently, the tube-like structural substance was subcutaneously implanted in 20 nude mice, which were regarded as an experimental group. And then, polyglicolide acid alone was subcutaneously transplanted in the rest 6 nude mice, which was regarded as a control group.MAIN OUTCOME MEASURES: Gross observations of cell-material compound by 2 and 6 weeks after transplantation;HE staining and immunohistochemical staining.detection; expression of factor Ⅷ and α-smooth muscle actin.RESULTS: Twenty-six nude mice were included in the final analysis. ① Gross observation: At 2 weeks after implantation,both the experimental and control groups formed tubular structures, however, at 6 weeks after implantation, the tubular structure still remained in experimental group but not in the controls. ② Histological observation and immunohistochemical detection: The histological examination of the engineered vessel showed that at 2 weeks, the vessels in both group contained mainly undegraded PGA fibers, while at 6 weeks, the PGA fibers were almost completely degraded in both groups and in the control group only fibrous-like tissue formed. Contrastly, in experimental group a typical vascular structure formed, Masson's trichrome stain, which stains collagen green, smooth muscle fibers red and cells purple, showed significant amounts of stainable collagen and smooth muscle fibers in the wall of the engineered vessel, furthermore,immunohistochemistry examination revealed that there were an endothelial cell layer formed in the inner surface of the engineered vessel which was confirmed by positive staining of yon Willebrand factor, meanwhile, the smooth muscle cells in the wall of the engineered vessel were confirmed by the positive staining of smooth muscle α-actin.CONCLUSION: The subcutaneous implantation of polyglicolide acid cocultured with vascular endothelial cells and smooth muscle cells may form vessels, which are similar to normal vascular histological structure.

OBJECTIVEElectrolytic detachable coils (EDC) have been the main embolic materials for intracranial aneurysms. Liquid aneurysmal embolic materials represented by cellulose acetate polymer (CAP) are still in controversy. In this research, the embolization results and pathological reactions after embolization of canine aneurysmal models with EDC or CAP were observed and compared.

METHODSThe canine aneurysmal models constructed by anastomosis of venous pouches were randomly grouped. The aneurysms were respectively occluded with CAP and electrolytic detachable coils that was named by Wu electrolytic detachable coil (WEDC) and made by us. Angiogram follow-ups were performed at 24-hour, 2-week, and 2-month after embolization. The occluded aneurysms were dissected in each stage for light microscopic, electron microscopic, and histochemical research.

RESULTSThe effect of embolization was significantly better with WEDC than that with CAP (chi2 = 5. 56, P < 0.05). Post-embolized complications such as aneurysm rupture and stenosis of parent arteries could only be found in CAP group. Pathological research showed that CAP mass could packed the aneurysms more densely than coils. Acute chemical damage of aneurysmal wall and inflammatory cell infiltration was prominently found in early stage after CAP-embolization. Organization of thrombus inside aneurysms and formation of endothelial tissue over the orifices of aneurysmal necks could be found in both groups 2 months after embolization. But parts of coils might be exposed outside endothelial layer.

CONCLUSIONSEDC are still the most safe, efficient, and reliable instruments to embolize aneurysm. CAP should be improved further to solve the problem of strong chemical corrosion and difficulty in control before it is widely used.

Animals ; Biocompatible Materials ; Cellulose ; analogs & derivatives ; Dogs ; Embolization, Therapeutic ; instrumentation ; methods ; Female ; Follow-Up Studies ; Intracranial Aneurysm ; therapy ; Male ; Random Allocation ; Tungsten

Objective To investigate the mechanism of matrix metalloproteinase(MMP)-9 involved in epithelial mesenchymal transformation(EMT)in chronic sinusitis(CRS).Methods The expression of MMP-9 from polypoid middle turbinate tissue was detected by immunohistochemical staining qPCR and Western blot assay in 42 patients with CRS and 8 patients underwent septoplasty.Primary human nasal epithelial cells HNEpc were cultured in vitro and divided into the control group,the TGF-β1 group(5 μg/L TGF-β1 intervention)and the TGF-β1+si-MMP-9 group(transfected with si-MMP-9 and 5 μg/L TGF-β1 intervention).The expression of MMP-9 was detected by cell immunofluorescence staining.Expression levels of TGF-β1,MMP-9 and EMT-related proteins E-cadherin,vimentin and α-SMA were detected by Western blot assay.Results(1)The positive expression rate of MMP-9 was significantly higher in the nasal mucosa of CRS with nasal polyps(CRSwNP)group(54.5%,12/22)than that of the CRS without polyps(25.0%,5/20)group and the control group(12.8%,1/8).The relative expression levels of MMP-9 mRNA and protein in nasal mucosa were higher in the CRSwNP group than those in the CRSsNP group and the control group(P＜0.05).(2)Compared with the control group,the expressions levels of TGF-β1,MMP-9,vimentin and α-SMA were increased in the TGF-β1 group,while the expression of E-cadherin was decreased(P＜0.05).Compared with the TGF-β1 group,expression levels of TGF-β1,MMP-9,vimentin and α-SMA were decreased in the TGF-β1+si-MMP-9 group,and the expression of E-cadherin was increased(P＜0.05).Conclusion The expression of MMP-9 is increased in CRS patients,which may be involved in the development of CRS through the regulation of EMT.

Objective To study the effect of magnetic rewarming on the morphology and biomechanical properties of vitrified umbilical artery. Methods The vitrified umbilical artery was rewarmed by magnetothermal method and traditional water bath. The temperature distribution and stress in the solution system were analyzed, and the rewarming effect was evaluated by tissue staining and mechanical test. Results Compared with water bath rewarming, the temperature gradient and thermal stress generated by magnetic rewarming were smaller, which could effectively reduce the thermal stress damage during the rewarming stage and achieve rapid and uniform rewarming. Magnetic rewarming could effectively avoid umbilical artery fractures and micro-cracks. After rewarming, the extracellular matrix, collagen fibers, elastic fibers and muscle fibers of the umbilical artery were evenly distributed, which preserved the macro and micro structures of the umbilical artery. The umbilical artery showed different degrees of hardening after water bath and magnetic rewarming, but the elastic modulus and limit stress of the latter were not significantly different from those of fresh umbilical artery, and the latter had unidirectional stretching characteristics similar to that of fresh umbilical artery, showing good elasticity and toughness. Conclusions Compared with water bath rewarming, magnetothermal method can effectively reduce the damage of rewarming stage, ensure the macroscopic, microscopic structure integrity of umbilical artery and better biomechanical properties. The research findings provide important references for cryopreservation of large tissues or organs such as umbilical artery.

Objective: To study the epidemiological characteristics of tsutsugamushi disease, and to confirm the existence of the disease's epidemic foci in Taizhou. Methods: From 2013 to 2014, Dongxing town hospital and Xingqiao town hospital were selected as specimen collection sites in Jingjiang city. Blood samples (5 ml) were collected from 40 patients with acute tsutsugamushi disease. A total of 59 rodents were captured with cage night method in the survey sites at 5, 7, 9, 10, and 11 months in 2013, from which, the spleen, liver, and kidney specimens were selected. Chigger mites were captured by small blackboard method and from the ears of the captured rodents. A total of 226 small blackboards were laid, 27 mites were captured, and the samples were grounded into suspension. Nested-polymerase chain reaction and cell and tissue culture techniques were used to test the specimen from the probable patients, host animals and chigger mites. Results: Among the 40 acute tsutsugamushi disease blood samples, 29 were found to meet the test requirements, 17 were positive for orientia tsutsugamushi nucleic acid with 59% of the positive rate, and 1 stran orientia tsutsugamushi was isolated. 59 rats were captured and the density of mice was 5.5%. Among them, there were 26 Mus musculus (2.4%), 18 Rattus flavipectus (1.7%) and 15 Smelly shrew (density 1.4%). 1 Smelly shrew was tested positive for orientia tsutsugamushi nucleic acid, and the negative results were found in the other rodent specimens. 27 Chigge mites were collected by small blackboard method and the density of mites was 0.12 for each blackboard, among which 3 larvae and 24 nymphs were found. 33 Chigger mites were collected from the ears of 3 Smelly shrew, and the density of the mite was 11 per mouse. All the captured Chigger mites were identified as Leptotrombidium scutellare and 1 group of specimens of Chigger mites from the external environment were positive for orientia tsutsugamushi nucleic acid. Conclusion There was a high density of mice in the epidemic area from May to November and the species of the chigger mites were Chigger mites in Taizhou. The nucleic acid of the oriental tsutsugamushi was detected in the patients with acute scrub typhus, rodents and vectors. According to the above-mentioned results, it was considered that the scrub typhus epidemic area of Taizhou city has the natural foci of scrub typhus.

Objective To explore the intervention effect and molecular mechanism of Xinkang Granules on inflammatory factors in rats with chronic heart failure based on cGAS/STING signaling pathway.Methods SD rats were randomly divided into normal group and modeling group.The chronic heart failure model was established by intraperitoneal injection of Doxorubicin Hydrochloride.After successfully modeling,the rats were further divided into model group,Valsartan group and Xinkang Granules group.The model group was treated with distilled water every day,the Valsartan group was treated with Valsartan solution every day,and the Xinkang Granules group was treated with Xinkang Granules every day,all given for 4 consecutive weeks.Echocardiography was used to detect cardiac function,the pathological changes of myocardium were detected by hematoxylin-eosin staining(HE),the ultrastructural changes of myocardium in each group were observed by transmission electron microscope,and the contents of interleukin-1β(IL-1β)and interleukin-6(IL-6)in serum were detected by enzyme-linked immunosorbent assay(ELISA).The mRNA expression levels of mitochondrial transcription factor A(TFAM),cyclic guanosine monophosphate-adenylate synthase(cGAS),interferon-stimulated gene(STING)and IL-6 in myocardial tissue of rats in each group were detected by real-time fluorescence quantitative method(qPCR).The protein expressions of cGAS and STING in rat myocardial tissue were detected by immunohistochemical method.Results Compared with the blank group,the rats in the model group had significant inflammatory cell infiltration and inflammatory edema in myocardial tissue,their cardiac function was significantly reduced(P＜0.05,P＜0.01),and serum inflammatory factors were significantly increased(P＜0.01).The mRNA expression of TFAM in myocardial tissue was significantly reduced(P＜0.01),the mRNA expressions of IL-6,cGAS,and STING were significantly increased(P＜0.01),and the protein expressions of cGAS and STING in the myocardial tissue were significantly increased(P＜0.01).Compared with the model group,the cardiac function of the rats in the Xinkang Granules group was significantly improved(P＜0.05,P＜0.01),the inflammatory infiltration of myocardial cells was reduced,the expression of serum inflammatory factors was significantly reduced(P＜0.01),the mRNA expression of TFAM in myocardial tissue was significantly increased(P＜0.05),and the mRNA expressions of IL-6,cGAS,and STING were significantly decreased(P＜0.01),the protein expressions of cGAS and STING in myocardial tissue were significantly decreased(P＜0.01).Conclusion Xinkang Granules can reduce the expression of inflammatory factors and improve cardiac function in rats with chronic heart failure.Its mechanism may be related to inhibiting the cGAS/STING signaling pathway.

The occurrence of benign prostate hyperplasia(BPH)was related to disrupted sex steroid hormones,and metformin(Met)had a clinical response to sex steroid hormone-related gynaecological disease.How-ever,whether Met exerts an antiproliferative effect on BPH via sex steroid hormones remains unclear.Here,our clinical study showed that along with prostatic epithelial cell(PEC)proliferation,sex steroid hormones were dysregulated in the serum and prostate of BPH patients.As the major contributor to dysregulated sex steroid hormones,elevated dihydrotestosterone(DHT)had a significant positive rela-tionship with the clinical characteristics of BPH patients.Activation of adenosine 5'-monophosphate(AMP)-activated protein kinase(AMPK)by Met restored dysregulated sex steroid hormone homeostasis and exerted antiproliferative effects against DHT-induced proliferation by inhibiting the formation of androgen receptor(AR)-mediated Yes-associated protein(YAP1)-TEA domain transcription factor(TEAD4)heterodimers.Met's anti-proliferative effects were blocked by AMPK inhibitor or YAP1 over-expression in DHT-cultured BPH-1 cells.Our findings indicated that Met would be a promising clinical therapeutic approach for BPH by inhibiting dysregulated steroid hormone-induced PEC proliferation.