Objective To evaluate the character of the collagen-chitosan-chondroitin sulfate scaffold seeded with rat adipose tissue-derived stromal cells. Methods A dipose tissue were harvested from 6 weeks old Wistar rats and the stromal cells were harvested by type Ⅰ collagenase and then cultured in vitro. Type Ⅰ collagen was fully mixed with chitosan, freeze-dried and cross-linked with chondroitin sulfate, then freeze-dried again and sterilized by ethylene oxide. The pore diameter, water content, porosity of the scaffold were tested. The adipose tissue-derived stromal cells were digested, seeded into the plates, scaffold, and cen-trifuged into pellet, and then induced into cartilage. MTT detection for cell proliferation was done. After 3 weeks, the cell morphology, and cell proliferation and adhesion were observed, and chondrngenic differenti-ation was also analyzed. Results The pore diameter, water content, porosity tested for the scaffold showed an appropriate form. Cell proliferation showed faster in the scaffold and pellet culture system after 5 day, there was still cell proliferation in the scaffold system after 14 days but no obvious changes in the pellet cul-ture system; ceils on the scaffold proliferated densely showed by histological staining, but there was a scaf-fold structure residues in the inner layer. The finding of type Ⅱ immunohistochemistry stain showed that cells express strong positive for type Ⅱ collagen in the scaffold and pellet culture system whereas it was weakly positive in the plate culture system; the specific mRNA for cartilage, type Ⅱ collagen, aggrecan and SOX-9 were expressed in all three systems showed by RT-PCR, but type X collagen was expressed continu-ously in the plate culture system and expressed after 21 days in the pellet culture system, whereas it was not detected in the collagen-chitosan-chondroitin sulfate scaffold system. Conclusion The parameters of the collagen-chitosan-chondroitin sulfate scaffold were suitable in our study. The results suggested that it can promote the adipose tissue-derived stromal cells proliferation and chondrogenic differentiation better than the plate and pellet culture systems and maintain the phenotype of chondrocytes well; it is the optimal choice for cartilage tissue engineering in the future.

BACKGROUND: The implanted cartilage calls can synthesize cartilage matrix as cartilage in cartilage tissue enginsedng, and the density of implanted cells is the key point.OBJECTIVE: To evaluate the effect of cell seeding concentration on the chondrogenic differentiation of the adipose dadved sromal cells (ADSCs).DESIGN, TIME AND SETTING: The in vitro cellular-scaffold observation was performed at the cytobiological laboratory of China Medical University from November 2007 to July 2008.MATERIALS: Six male SD rata with clean grade were supplied by the Experimental Animal Center of China Medical University.METHODS: Totally 5 g/L type ; collagen solution and 20 g/L chitosan was mixed in a mould with volume ratio of 7:3, after lyophillization, it was cut into pieces with 5 mm ～ 5 mm x 2 mm, followed by crosslinking with ethanol contained of 2% chondroitic acid at room temperature. After washing with double distilled water and freeze drying, the chitosan-collagen-chondroitin sulfate copolymar matrices scaffolds were harvested. ADSCs isolated from rat inguinal fat pads were digested with collagenase and trypsase. The prepared scaffolds were randomly divided into 3 groups, and the third passage cells with density of 2×10 9/L,2×10 109/L, and 2×10 11/L were seeded into chitosan-coflagen-chondroitin sulfate scaffolds, and cultured in chondrogenic medium for 3 weeks.MAIN OUTCOME MEASURES: The expression of cartilage specificity gene was detected by hematoxylin-eosin staining, type Ⅱ collagen immunohistochemical staining and RT-PCR.RESULTS: Hematoxylin-eosin staining showed that after 3 weeks of culture, the cell proliferated and differentiated well, especially in 2x101～/L group, more extrocelluer matrices were produced and cartilage lacuna-structure could be seen. The type Ⅱ collagen was positive expressed in each group, which showed a gradually increasing tendency with the cell seeding concentration increasing. RT-PCR showed that the expression of proteoglycen and type Ⅱ collagen mRNA were slowly increased. However the expression of Ⅹ collagen mRNA was decreased with increasing cell seeding concentration.CONCLUSION: The chitosan-collagen-chondroitin sulfate copolymer matrices can provide an appropdate environment for the generation of cartilage-like tissues and high call seeding concentration of 2×1010/L facilitate ADSCs to differentiate into cartilage.

Background and purpose: The prognosis of completely resected stage ⅢA(N2) non-small cell lung cancer (NSCLC) remains a significant concern. The 5-year overall survival (OS) rates range from 10% to 30%. This study aimed to analyze the patterns of first failure in completely resected stage ⅢA(N2) NSCLC and to assess the actuarial risk of developing metastasis at different sites and to guild standard clinical practice. Methods: Patients withⅢA(N2) NSCLC who had undergone radical surgery in our hospital from Jan. 2005 to Jul. 2012 were retrospectively reviewed. The progression-free survival (PFS), the OS, patterns of first failure, the actuarial risk were analyzed. The cumulative incidence of first failure was determined using the Kaplan-Meier analysis. Results: Among 357 patients who met the eligibility criteria with completely resected stage ⅢA(N2) NSCLC, 5-year OS was 36.9%. There were 284 (77.6%) patients experiencing disease failure: 61 with local failure, 197 with local and distant failures, and 26 patients with local recurrence as the first failure. Brain, bone and lung were the main sites of distant failure as the first failure, while brain was the most common site. There were 67 patients developing brain metastases (BM) as the first site of failure. The median time of local failure as the first site of failure was 13.6 months, and the time to develop distant recurrence was 15.1 months. 92.5% BM developed in 3 years after the complete resection. Conclusion: As the first failure, the rate of distant failure was much higher than that of local failure in completely resected stage ⅢA(N2) NSCLC. Brain was the most common site of distant failure as the first failure. These results can be helpful in guiding standard clinical practice and evaluating the outcome of comprehensive treatment.

Purpose To investigate the clinicopathologic features of cervical adenoid basal cell carcinoma. Methods Clinical and pathological data were analyzed in 12 cases of cervical adenoid basal cell carcinoma and the relevant literatures were reviewed. Results The age of 12 cases of with cervical adenoid basal cell cancer patients ranged from 36~70 years ( mean:53. 3 years) , with a medi-an age of 51. 5 years. Amongst 12 patients who contained no gross definite lesion, 5 patients (41. 7%) had the symptoms of vaginal bleeding. 8 cases of patients underwent hysterectomy, while the other four received cervical conization. Cervical intraepithelial neopla-sia ( CIN) lesions were observed in all 12 cases. The tumor cells were small and uniform, with dark oval nuclei without conspicuous nucleoli and scanty cytoplasm. Tumor infiltrated into the stroma in nests and cords. Glandular differentiation within tumor nests were seen in 12 cases, including two cases of squamous cell differentiation accompanied by partial and minor interstitial edema. Peripheral palisading cells around tumor nests were also found. The infiltrating depth of tumor ranged from 0. 5~10 mm with the average 4. 12 mm. Cancer embolis were available in 2 cases with no lymph node metastasis. 10 cases followed up for 3~78 months with no recur-rence and metastasis, but 2 cases were lost. Conclusion Adenoid basal cell carcinoma is a rare uterine cervical tumor found in post-menopausal women, which are often accompanied with CIN. This tumor has a favorable prognosis and should be clearly separated from adenoid cystic carcinoma and other tumors which also have infiltrative growth pattern.

Objective To explore the relationship between the protective effect of hydrogen sulfide (H2 S ) postconditioning against myocardial hypoxia-reoxygenation and the dynamics of actin. Methods Adult rat cardiomyocytes were isolated and the same hatch cells were divided into 4 groups (n =3):normal group (group N),hypoxia-reoxygenation group (group HR),ischemia postconditioning group (group IPTC)and H2 S postconditioning group (group S).The four groups were divided into two sub-groups with or without cytochalasin D (CyD).At the end of reoxygenation,F-actin/G-actin,intracellular calcium ion concentration (Ca2+ )and pH value were detected with laser scanning confocal microscopy, meanwhile the level of p-p38MAPK was detected with Western blot.Results The fluorescence intensity of F-actin/G-actin of group HR,group IPTC and group S were significantly higher than that of group N (P <0.05).The fluorescence intensity of Ca2+ of group HR was higher than that of group N,group IPTC and group S (P <0.05);The intensity of Ca2+ of all group with CyD treatment was higher than those without (P <0.05);The fluorescence intensity of pHi of group N and group HR with CyD treatment was higher than those without;The fluorescence intensity of pHi of group IPTC and group S was lower than those without;The flu-orescence intensity of pHi of group HR was lower than that of group N,group IPTC and group S (P <0.05), respectively;the pHi of group N and group HR sub-group with CyD treatment was higher than those without correspondingly (P <0.05),however,the pHi of IPTC and S sub-groups were lower than their corresponding groups (P <0.05).The level of p-p38MAPK in group HR was significantly higher than those of other groups (P <0.05);there was no difference among groups N,IPTC and S.Conclusion Hydrogen sulfide postcondi-tioning could promote F-actin to remodel and stabilize the cellular environment.Hypoxia-reoxygenation pro-motes the phosphorylation level of p38MAPK,which could be surpressed by H2 S postconditioning.

OBJECTIVETo study the effect of electro-acupuncture (EA) at Neiguan (PC6) and Lieque (LU7) on the expression of protein kinases in cardiomyocytes of myocardial ischemia (MI) rats.

METHODSHealthy male SD rats were randomly divided into the control group, the model group, the Neiguan point group, the Lieque point group, and the non-meridian non-acupoint group, 10 in each group by random digit table. The MI rat model was established by injecting isoprenaline hydrochloride (85 mg/kg). EA at Neiguan (PC6), Lieque (LU7), and non-meridian non-acupoint were respectively performed. Changes of the expression of protein kinases [such as protein kinase A (PKA), protein kinase C (PKC), protein kinase G (PKG)] in rat cardiomyocytes were observed using Western blot.

RESULTSCompared with the control group, expression levels of PKA, PKC, and PKG increased obviously in the model group (P < 0.01). Compared with the model group, expression levels of PKA, PKC, and PKG decreased in the Neiguan point group and the Lieque point group (P < 0.01, P < 0.05). Expression levels of PKA decreased in the non-meridian non-acupoint group (P < 0.01). Compared with the Neiguan point group, expression levels of PKA, PKC, and PKG increased in the non-meridian non-acupoint group and the Lieque point group (P < 0.01, P < 0.05). Compared with the Lieque point group, expression levels of PKA, PKC, and PKG increased in the non-meridian non-acupoint group (P < 0.01, P < 0.05).

CONCLUSIONEA at Neiguan (PC6) and Lieque (LU7) could decrease protein expression levels of PKA, PKC and PKG in rat myocardial cells, and the effect of acupuncture at Neiguan (PC6) was better than that obtained by EA at Lieque (LU7).

Acupuncture Points ; Acupuncture Therapy ; Animals ; Coronary Artery Disease ; Cyclic AMP-Dependent Protein Kinases ; Electroacupuncture ; Male ; Meridians ; Myocardial Ischemia ; metabolism ; therapy ; Myocytes, Cardiac ; metabolism ; Plant Extracts ; Protein Kinases ; metabolism ; Rats ; Rats, Sprague-Dawley

Humans ; Male ; Middle Aged ; Pacemaker, Artificial ; adverse effects ; Tricuspid Valve Stenosis ; etiology

Astragalus membranaceus var. mongholicus and Hedysarum polybotrys belong to different genera, but have similar drug efficacy in traditional Chinese medicine theory, and H. polybotrys was used as the legal A. membranaceus var. mongholicus previously. In this study, similarities and differences between them were analyzed via their ITS/ITS2 fragments information. The ITS (internal transcribed spacer) regions were amplified using polymerase chain reaction and then sequenced in two-way. The alignment lengths of ITS regions were 616 bp, in which 508 loci were consistent, and 103 loci were different, accounting for 82.47% and 16.72% of the total ITS nucleotides in length, respectively. As genotype determines phenotype, 1HNMR-based metabolomic approach was further used to reveal the chemical similarities and differences between them. Thirty-four metabolites were identified in the 1H NMR spectra, and twenty-seven metabolites were the common components. Amino acids, carbohydrates and other primary metabolites were similar, while a large difference existed in the flavonoids and astragalosides. This study suggests that A. membranaceus var. mongholicus and H. polybotrys show similarities and differences from molecular and chemical perspectives, which has laid a foundation for elucidating the effective material basis of drug with similar efficacy and resources utilization.
Astragalus membranaceus ; chemistry ; genetics ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drugs, Chinese Herbal ; chemistry ; Fabaceae ; chemistry ; genetics ; Flavonoids ; chemistry ; Metabolome ; Metabolomics

Heart Neoplasms ; pathology ; Heart Ventricles ; pathology ; Humans ; Infant, Newborn ; Male ; Rhabdomyoma ; pathology

Objective: To investigate the status of electronic cigarette use among adolescents in Ningxia Hui Autonomous Region, and to provide evidence for tobacco control in adolescents. Methods: Based on the 2019 National Youth Tobacco Epidemic Monitoring Program, multistage proportional sampling method was used to select middle school students from Ningxia Hui Autonomous Region. A questionnaire revised by Chinese CDC was used to collect the general information, the cognition and use of electronic cigarettes, and the access to advertising of electronic cigarettes and related products. Results: Totally 9 019 questionnaires were distributed, 8 401 valid ones were recovered, and the response rate was 93.2%. The rates of electronic cigarette use and attempt among students were 4.3% and 13.4%. The rates of electronic cigarette use and attempt in male students were 7.7% and 22.9%, which were higher than that in female students (0.8% and 3.8%, P<0.05) . The rates of electronic cigarette use and attempt varied in different schools ( P<0.05 ), which were higher in vocational high school students ( 11.5% and 26.8% ). Among 246 students who used electronic cigarettes, 30.1% did not thought electronic cigarettes contained nicotine, while 60.2% did not know whether electronic cigarettes contain nicotine. In the past 30 days, 27.0% of the students had seen the advertisements of electronic cigarettes and related products, mainly through TV, store, supermarket, convenience store, grocery store, electronic cigarette experience store or retail store. Conclusions The rates of electronic cigarette use and attempt among adolescents in Ningxia Hui Autonomous Region are 4.3% and 13.4%. Boys and vocational high school students have higher rates. Students generally know electronic cigarette and have more access to it.