BACKGROUND:Effects of different oxygen partial pressures on cytokine secretion of human adipose-derived stem cells have been differently reported. These differences may be caused by varying oxygen partial pressures.
OBJECTIVE:To investigate the influence of different oxygen partial pressures on cytokines secreted from human adipose-derived stem cells.
METHODS:Human adipose-derived stem cells were cultured in vitro and identified by its immunophenotype. Human adipose-derived stem cells were divided into five groups and cultured under different oxygen partial pressure conditions (1%, 3%, 5%, 10%, 21%) for 24 hours, respectively. With quantitative real-time PCR and enzyme linked immunosorbent assay, the secretion of cytokines, vascular endothelial growth factor, hepatocyte growth factor, nerve growth factor, keratinocyte growth factor, from human adipose-derived stem cells were analyzed on the gene and protein levels.
RESULTS AND CONCLUSION:Human adipose-derived stem cells were positive for CD71, CD73, CD90, CD105 and negative for CD34, CD45, CD54, HLA-DR. From the aspect of gene level, hypoxia (1%, 3%O 2 ) promoted the expression of vascular endothelial growth factor and nerve growth factor from human adipose-derived stem cells (P<0.01), and significantly elevated the expression of hepatocyte growth factor (P<0.05);however, there was no significant influence on keratinocyte growth factor under hypoxia (P>0.05). Based on the protein level, protein secretion of hepatocyte growth factor and vascular endothelial growth factor from human adipose-derived stem cells was increased under hypoxia (P<0.01), but no changes occurred in nerve growth factor and keratinocyte growth factor. After cultured under hypoxic environment, human adipose-derived stem cells were promoted to express gene vascular endothelial growth factor, hepatocyte growth factor and nerve growth factor, as wel as to secrete protein keratinocyte growth factor and vascular endothelial growth factor.

Objective This study aimed at observing expression and clinical significance of metadherin in gastric adenocarcinoma and exploring the potentially regulating mechanism of metadherin in invation and migration of gastric cancer.Methods Expressions of metadherin and E-cadherin in primay lesion of gastric cancer were detected by immunohistochemistry and their correlation to clinicopathologic characteristics and prognosis were analyzed by Chi-square tests.Transwell assay and wound healing assay were applied for the ability of invasion and metastasis in gastric cancer cells.Then,the down-regulatied metadherin expression in MKN45 cells by RNA interference (siRNA) was carried out and furthermore,the regulation role of metadherin in epithelial-mesenchymal transition was analyzed also in invasion and migration of gastric cancer cells.Results The positive expression of metadherin was correlated to invading depth (P =0.029),lymph node metastasis (P =0.001),TNM stage (P =0.014) and inhibiting E-cadherin expression (P =0.001).The patients with positive metabherin shared poorer prognosis.Furthermore,the down-regulated metabherin in MKN45 cells would result in the increasing expression of E-cadherin,as well as decreasing expression of N-eadherin,Slug and Snail.At the same time,the abilities of invasion (P =0.027) and migration (P =0.008) of MKN45 cells was decreased.Conclusion metabherin induces EMT in metastasis of gastric adenocarcinoma via activating either Slug or Snail but not twist,which would result in the poorer prognosis.

AIM: To investigate the effect of sitagliptin on the autopaghy and the expression of extracellular matrix in mesangial cells induced by advanced glycation end products (AGEs).METHODS: The cells were divided into 5 groups: control group, AGE group, and sitagliptin (5, 10 and 20 μmol/L) groups.After 48 h, the cell viability was measured by MTT assay, and the content of collagen (Col) Ⅳ in the supernatant of the cell culture was detected by ELISA.The protein levels of beclin-1, adenosine monophosphate-activated protein kinase (AMPK), p-AMPK, p70S6K and p-p70S6K were determined by Western blot.RESULTS: Compared with control group, the viability and the expression of Col IV induced by AGEs in the cultured mesangial cells were significantly increased (P<0.01).Sitagliptin decreased the viability and the expression of Col IV induced by AGEs in the mesangial cells in a dose-dependent manner.AGEs significantly inhibited the protein levels of beclin-1 and p-AMPK, but significantly increased the protein level of p-p70S6K.Compared with AGE group, sitagliptin significantly reversed the above results in a dose-dependent manner.CONCLUSION: Autophagy may mediate the protective effect of sitagliptin on mesangial cells induced by AGEs.

Objective To report the outcome of emergency repair traumatized limbs by vascularized free tissue transplantation.Methods From April 1988 to August 2004,86 patients,58 men and 28 women,had undergone emergency vascularized free tissue transplantation to have their injured limbs repaired in 54 cases and the missing thumbs reconstructed in 32.The patients aged from 5 to 55 (mean 27.9) years. The transplants included latissimus dorsi myocutaneons flap,anterolateral femoral skin flap,medial crural skin flaps,dorsal pedal flaps,medial plantar flap,composite tissue mass of the discarded limbs and big toe skin- nail flap.The operations were performed 1 to 5 days after injuries.Results Postoperative vascular crises occurred in 8 cases and were all followed by exploration with successes in 5 cases while failure in 3.The total survival rate of the transplants was 96.5% (83/86).In this series all the patients were followed up for 1 to 16 years with a mean of 7.5 years only to reveal satisfying functional recovery in all the repaired limbs and an ex- cellent and good rate of 87.5% in the reconstructed thumbs.Conclusion Emergency vascularized free tis- sue transplantation is an effective way to repair a traumatized limb and to reconstruct a traumatically missing thumb.

OBJECTIVETo establish the steriod-induced osteoporosis model with the type of Kidney-Yin deficiency.

METHODSTotally 45 female Kunming mice were randomly divided into normal group,model group and Liuwei Dihuang pills(Chinese character: see text)group. The model was established by intramuscular injecting of Dexamethasone. Liuwei Dihuang pills (Chinese character: see text) group was administered orally with Liuwei Dihuang pills (Chinese character: see text). The signs and symptoms of mice were observed dynamically. All the animals were sacrificed at the end of the 6th weeks. The level of ACTH, cAMP, cGMP, TSH and E2 in serum were detected to evaluate deficiency of Kidney-Yin. Morphological changes and bone density were observed to evaluate osteoporosis.

RESULTS(1) Compared with control group, mice in model group appeared obvious Kidney-Yin deficiency symptoms, including hair dry, restlessness, excitability, hard stool, and yellow. (2) Compared with control group,the weight of mice in model group gained slower (P<0.01); the index of adrenal gland,liver and spleen decreased (P<0.01, P<0.01 ,P<0.01); the level of ACTH and TSH increased (P<0.01 ,P<0.01), the level of E2 decreased (P<0.01) and the ratio of cAMP/cGMP increased (P< 0.05). (3)Compared with control group,the bone density of lumbar vertebra and femur in model group were significantly decreased (P<0.01, P<0.05); HE staining revealed osteoporosis in model group mice. (4)However, the Liuwei Dihuang pills (Chinese character: see text) group can partly antagonize the inhibition of the HPA axis, alter the disordered sex hormone and the ratio of cAMP/cGMP, and reverse the osteoporosis partly.

CONCLUSIONthe model of osteoporosis with type of Kidney-Yin deficiency could be established by Dexamethasone intramuscular injection. With less interference, it wight be a stable and reliable modeling method for integration of disease and syndrome in TCM.

Animals ; Bone Density ; Dexamethasone ; toxicity ; Disease Models, Animal ; Female ; Kidney Diseases ; etiology ; Medicine, Chinese Traditional ; Mice ; Osteoporosis ; chemically induced ; Yin Deficiency ; complications

Objective To use CT and 3D reconstruction technique to locating mark of sustentaculum tali ,and measure the an‐gle & length of the specified point of the lateral wall of the calcaneus to sustentaculum tali for providing .To basis for the operation of calcaneus .Methods Thirty adult ankle wet specimens were chosen and scanned with CT ,and the dicom data were converted into the software to 3D‐reconstruction ,the observation and measurement to find new methods of locating mark of sustentaculum tali ;Forty adult calcaneus dried specimens were chosen and scanned with CT .The dicom data were converted into the software to 3D‐re‐construction ,using a workstation software measure the angle and length of the line which from the center of the sustentaculum tail to each point on the calcaneal lateral wall .Results First ,in the function of ankle joint position ,with the foot medial surface as “sag‐ittal reference plane” measurement ,the vertical line passing through the leading edge of medial malleols and the coronal line passing through the leading edge of sustentaculum tali intersect at a point .The vertical distance of the point to leading edge of medial malle‐ols was (34 .95 ± 2 .60) mm ,the length of sustentaculum tali was (21 .44 ± 1 .89) mm ,the height of sustentaculum tali was (10 .17 ± 1 .16) mm ,the anteversion of sustentaculum tali was (35 .31 ± 3 .73)° .Second ,in front of calcaneus ,upward angel of points G ,E were (28 .78 ± 3 .90 )° ,(29 .47 ± 2 .96 )° respectively .Frontward angle of points G ,E were ( - 19 .83 ± 4 .60 )° , ( - 11 .02 ± 4 .74)° respectively .Under the posterior articular surface ,upward angel of points C ,K ,I ,J were(2 .52 ± 2 .46)° ,(2 .92 ± 2 .28)° ,(14 .98 ± 2 .49)° ,(14 .38 ± 1 .90)° respectively .Frontward angle of points C ,K ,I ,J were(7 .32 ± 1 .66)° ,(19 .25 ± 1 .98)° , (10 .30 ± 2 .63)° ,(19 .33 ± 1 .97)° respectively .The length of screw was about 40 - 44 mm .The length of C point to the sustentacu‐lum tail was minimum ,and the length of G was maximum .Conclusion In the function of ankle joint position ,the sustentaculum tail can be located with the leading edge of medial malleols as a surface landmarks .The measurement of the angle and length of which from each point on the calcaneal lateral wall to sustentaculum tail by using CT .

Objective To evaluate the effects of rigorous surveillance and retroperitoneal lymph node dissection (RPLND) in the treatment of low-risk patients with clinical stage Ⅰ nonseminomatous germ cell testicular tumors (NSGCT) after radical orchiectomy.Methods The data of 71 patients with clinical stage Ⅰ NSGCT were analyzed retrospectively in Hunan Provincial Tumor Hospital,Xiangya Third Hospital of Central South University and Hunan Provincial People's Hospital between Feb,2001 and Apr,2012.Excluding lymphatic and vascular invasion,percentage of embryonal carcinoma＞50％ and increasing tumour markers (AFP/β-HCG) following orchiectomy,46 low-risk patients out of 71 patients with clinical stage Ⅰ NSGCT were selected and divided into rigorous surveillance group (30 cases) and RPLND group (16 cases) according to different therapeutic methods after radical orchiectomy.Univariate analysis was used to confirm variables associated with disease progression,and the disease free survival rates (DFSR) were compared by using Kaplan-Meier analysis.Results Five cases were lost,and 41 cases were followed up for an average of 61 months (range,15-147 months),with 58 months in rigorous surveillance group (range,19-147months) and 65 months in RPLND group (range,15-144 months).The survival rate was 100％ in 2 groups.The DFSR was 89％ (24/27) and 86％ (12/14),respectively,and there was no significant difference between the 2 groups (x2 =0.08,P=0.78).The DFSR was 83％ in patients with small amout of embryonal (percentage of embryonal carcinoma ＜ 50％),and 92％ in patients without embryonal carcinoma,and there was no significant difference between the 2 groups (x2=1.07,P=0.30).Also there was no significant difference between the patients less than 15 years and patients more than 15 years (x2=1.59,P =0.21).Conclusions There is no significant difference in recurrence rate and DFSR between rigorous surveillance group and RPLND group.Low-risk patients with clinical stage Ⅰ NSGCT may achieve satisfactory prognosis with surveillance after radical orchiectomy.

Experiment teaching is a most important item of college teaching, and plays a vital and unexchangeable role to train students for their ability to practice and to innovate. To construct a feasible and scientific examination system of experiment teaching, will significantly help deepen the reformation of experiment teaching and improve the teaching quality. So according to the request for cultivating qualified ap- plication person, we preliminarily constitute the valuation system throughout the whole course and in the final, of theoretic examination and practice examination, and combined with students’ self-valuation and valuation from teachers. In fact, the system works well with a perfect effect.

Objective To investigate the MRI features of intracranial solitary fibrous tumors/hemangiopericytomas (SFT/HPC),and to compare these findings with those of intracranial meningiomas.Methods The clinical features and MRI findings in 28 patients of intracranial SFT/HPC (SFT/HPC group)and 68 patients of meningiomas (meningiomas group) confirmed by operation and pathology were retrospectively analyzed.The indicators of two groups were compared.Results Shape of tumor,signal homogeneous,signal voids of vessel in tumor,hypointense signal nodules on T2WI and enhanded,cystic or necrosis in tumor,meningeal tail sign,changes of the nearby bone,sex,Ki-67％ level,blood lose in surgery had significant differences between SFT/HPC group and meningiomas group (all P＜0.05).Conclusion There are some differences between intracranial SFT/HPC and meningiomas.It is helpful in diagnosis and differential diagnosis through the comparative analysis of the imaging signs.

Objective:The purpose of this study is to investigate the expression of tumor stem cell marker aldehyde dehydrogE-nase 1 (ALDH1) in breast cancer and its clinical significance. Methods:The expression of ALDH1 protein was examined by immunohistochemical staining in 92 breast cancer tissues. The correlation analysis and diseasE-free survival analysis of patients was evaluated based on the clinical follow-up data. Results:Expression of ALDH1 protein had a significant correlation with progesterone receptor (PR) and cerb-B2 (P<0.05), but had no significant correlation with age, tumor size, clinical staging, and lymph node metastasis (P>0.05). The 2-year diseasE-free survival rate of AlDH1-positive patients was lower than that of ALDH1-negative patients (P<0.05). ALDH1-positive patients, who received CEF regimen chemotherapy and hormone therapy, had lower 2-year diseasE-free survival rate than that of ALDH1-negative patients (P<0.05). Conclusion:The decreased diseasE-free survival rate of ALDH1-positive patients is related with drug resistance. ALDH1 could be used as an independent factor for predicting the prognosis of breast cancer.