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The spider neurotoxin hainantoxin-IV(HNTX-IV), which is isolated from the crude venom of the spider Selenocosia hainana, can specifically inhibit the tetrodotoxin-sensitive(TTX-S) sodium channel, and can selectively inhibit Voltage-gated sodium channel(VGSC) Na
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Aim To explore the effect of salidroside on the learning and memory ability of mice under high altitude hypoxia. Methods Forty-eight C57BL/6J male mice were randomly divided into plain control group, plateau model group and salidroside group according to their body weight, with 16 mice in each group. The animals in each group were given prophylactic doses for three days and then rushed to a plateau with an altitude of 4 010 m. After one day of hypoxia exposure, Morris water maze was performed to test the learning and memory ability of mice; malondialdehyde(MDA), hydrogen peroxide(H
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Hie high altitude hypoxic environment affects the pharmacokinetic process of rlnjgs by changing the body's gastrointestinal emptying rate, organ blood flow, drug plasma protein binding rate, dnjg metabolizing enzymes and transporter expression.Epilepsy is a brain disease that requires long-term medication.Most anti-epileptic drugs have a low therapeutic index and a narrow range of effective blood drug concentrations.'Ilierapeu- tic dnjg monitoring (TDM) is commonly used clinically to find the best individualized medication method for antiepileptic dnjgs.rI1iis article summarizes the commonly used anti-epileptic dnjgs and their treatment windows in clinical practice, and analyzes the influence of the pharmacokinetics of anti-epileptic dnjgs in the high altitude hypoxic environment, so as to provide reference for the clinical use of anti-epileptic drugs at high altitude.
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Aim To investigate the effects of altitude hypoxia on serum sodium valproate eoncentration and eerebral blood distribution.Methods Male mice were divided into control group and plateau group.Each group was given sodium valproate orally and intrave¬nously, respectively.UFLC-MS/MS was used to deter¬mine the concentration of sodium valproate in plasma and brain, and Western blot was used to detect the ex¬pression of P-gp in BBB.Results Compared with the control group, the ratio of brain/blood drug concentra¬tion in plateau group was up-regulated by 44.0% , 57.9% , 176.8% and 184.5% at 10, 30, 60 and 120 min, respectively.The ratio of brain/blood drug con-centration increased by 33.9% , 50.6% and 125.6% at 60 min, 120 min and 240 min in plateau group, re¬spectively.Compared with the control group, the ex¬pression of P-gp protein in BBB of mice in altitude group was significantly down-regulated by 58.46% (P < 0.05 ).Conclusions Compared with the control group, the brain/blood drug concentration ratio of val¬proic acid increases in high altitude hypoxia environ¬ment.Meanwhile, it is found that P-gp expression lev-el decreased in the brain mierovessels of mice under high altitude hypoxia environment, and the cerebral and blood distribution of valproic acid in mic increases in high altitude hypoxia environment.
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<b>Objective::To study whether Sanhuang Xiexintang (SHXXT) can restore endothelial function by inhibiting the activation of NOD-like receptor protein 3 (NIRP3) induced by 7-ketocholesterol (7-keto) in vascular endothelial cells. <b>Method::The aortic rings of mice were cultured in normal group, model (7-keto) group, SHXXT groups (1%, 2% and 5% drug-containing serum). Vasodilation function of mice was observed. Microvascular endothelial cells were cultured according to the above experimental groups, and NIRP3 inhibitor isoglycyrrhizin (ISO) group, was also set. Western blot was used to detect the expressions of endothelial nitric oxide synthase (eNOS), NIRP3, cysteinyl aspartate specific proteinase-1 (Caspase-1), interleukin-1<italic>β</italic> (IL-1<italic>β</italic>) protein. In addition, nitric oxide (NO) quantitative kit was used to detect the concentration of NO. <b>Result::Compared with the normal group, the endothelium-dependent vasodilation function of vascular rings was significantly reduced in model group (<italic>P</italic><0.01), and the drug group significantly restored the endothelium-dependent vasodilation function in a concentration-dependent manner (<italic>P</italic><0.05, <italic>P</italic><0.01). Meanwhile, microvascular endothelial cells were also studied. Compared with the normal group, the content of eNOS protein in the model group decreased (<italic>P</italic><0.05), while the concentration of NO decreased significantly (<italic>P</italic><0.01). After treatment with SHXXT serum, eNOS and NO could be restored, with significant differences in the concentration of NO with 5% (<italic>P</italic><0.05) and 10% (<italic>P</italic><0.01) SHXXT serum. At the same time, the expressions of NIRP3 (<italic>P</italic><0.05), cle-Caspase-1 activation (<italic>P</italic><0.01) and IL-1<italic>β</italic> production (<italic>P</italic><0.01) in endothelium were significantly increased under 7-keto stimulation, and the SHXXT serum could significantly inhibit the expression and activation of relevant proteins. Subsequently, endothelial cells were treated with NIRP3 inhibitor ISO. Compared with the model group, eNOS expression increased, and NO concentration increased significantly (<italic>P</italic><0.01) after treatment with ISO, but ISO had no synergistic effect on SHXXT serum. <b>Conclusion::SHXXT can improve endothelium-dependent vascular dysfunction induced by 7-keto, which is achieved by NO signaling pathway mediated by inhibiting the activation of endothelial NIRP3-related proteins.
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Postpartum hemorrhage,a frequent emergency in obstetrics,is the first reason for maternal mortality in China and worldwide.The contributors to adverse outcome included delayed treatment because of underestimation of blood loss,lack of blood products,lack of treatment algorithms,lack of knowledge and training,insufficient interdisciplinary communication and inadequate management.Therefore,it is essential to improve the management of medical institutions and set up a protocol for emergency situations.Meanwhile,the key point in rescue postpartum hemorrhage is the teamwork with efficient training and cooperation.This article made a review on the management of postparttum hemorrhage,establishment of emergengy protocol and risk assessment and team cooperation based on guidelines and recommendations published by main authorities and the latest studies.The goal of the article is to provide some ideas to reduce the frequency of severe hemorrhages after delivery and improve maternal outcomes.
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OBJECTIVE@#To investigate whether cell preservation solution can prolong the survival time of leukemia cells and increase the survival rate, so as to improve the detection rate of central nervous system leukemia.@*METHODS@#Kasumi cells were added into cerebrospinal fluid (CSF) supernatant with or without cell preservation solution to compare cell viability and biological characteristics at different time point. Wright Giemsa staining was used to compare cell morphology; cell counting, CCK-8 method, and trypan blue staining were used to compare the cell number, and flow cytometry was used to compare the cell viability. The expression of AML-ETO tumor fusion gene was detected by fluorescence quantitative RT-PCR.@*RESULTS@#At different time points (8 h and 24 h), the survival, molecular biological characteristics and RT-PCR result of the cells in CSF with cell preservation solution were significantly better than those in normal cerebrospinal fluid.@*CONCLUSION@#Cell preservation solution can effectively improve the survival time and survival rate of leukemic cells, thereby increase the detection rate of CNS leukemia.
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Humans , Central Nervous System Neoplasms , Core Binding Factor Alpha 2 Subunit , Leukemia , RUNX1 Translocation Partner 1 ProteinABSTRACT
Objective:To study the perceived organizational support,organizational approval impact on job engagement process and influence mechanism of non-establishment staff in public hospital under the background of China's personnel sys-tem,and provide reference for hospital managers to take measures to promote the staff's job involvement. Methods:650 non-es-tablishment staffs from three tertiary public hospitals in each of the six cities of Sichuan province were surveyed by issuing or-ganizational support questionnaires,organization approval scale and work input scale. Descriptive analysis,correlation analy-sis,regression analysis were performed with the help of SPSS 19.0,a three-variable structural equation model was established by using AMOS17.0. Results:The average score of organizational support of non-establishment staffs in public hospitals was (2.80 ±1.05),the organizational approval scale was(3.73 ±1.03)and job engagement was(2.99 ±1.58);and there was a sig-nificant positive correlation (P<0.01) between organizational approval and job engagement. It was also revealed that per-ceived organizational support had a direct effect on job involvement,and indirect effect on organizational approval as a partial mediation variable. Conclusion:Public hospitals have a low level of perceived organizational support,job involvement of non-establishment staffs,and public hospital administrators can promote the job engagement of non-establishment staffs in public hospital by improving the sense of organizational support and organizational identification.
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[Objective] We assessed the associations between protein intake and T2D in different dietary patterns.[Methods] We conducted a population-based cross sectional study of 5 956 women and 2 298 men in middle-aged and elderly population in the community of Guangzhou city,Guangdong province.Cluster analysis was used to category subjects into mutually exclusive group by major sources of protein.Logistics regression analysis and substitution models were used to estimate T2D risks according to protein intake.[Results] The mean age of subjects was 55.51 year.Subjects were divided into four dietary protein food patterns (coarse cereals,red meat,refined grain,and sea food).Overall,extreme quantile of total protein intake was significantly and positively associated with T2D [odds ratio 1.58 (95％ CI 1.25,2.02)].In subgroup analysis by dietary patterns,extreme quantile of total protein intake was also positively related to T2D in the "red meat" [3.00(1.36,7.01)] and "refined grain" [2.25 (1.40,3.61)] dietary pattern group.However,this association was revered in the "coarse cereals" group [0.48 (0.23,0.97)];and protein intake was not related to T2D in the "seafood" group.[Conclusions] The association between protein intake and T2D varies by dietary pattern.Dietary pattern should be considered into the recommendation of protein intake for diabetes prevention.
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Objective To observe the changes in the circumpapillary thickness parameters measured by optical coherence tomography (OCT) in patients with optic disc edema (ODE).Methods Together 36 patients (36 eyes) diagnosed with ODE from October 2012 to October 2015 were included in the study,and 51 age-and gendermatched control subjects (51 eyes) were recruited from among healthy individuals with no history or evidence of current ocular disease.All the subjects were examined for visual acuity,slit lamp,fundus and fundus photography,and all the eyes underwent OCT scan to detect the circumpapillary retinal nerve fiber layer thickness (cpRNFLT) and circumpapillary total retinal thickness (cpTRT).Results After OCT scan,25 ODE patients (5 mild,14 moderate and 6 severe patients) and 30 patients (5 mild,15 moderate and 10 severe patients) were in line with the requirements of cpRNFLT analysis and cpTRT analysis respectively;there were 50 controls meeting the requirements of this study.There was no significant difference in age and sex between ODE patients and the controls (all P ＞0.05).The mean cpRNFLT and cpRNFLT in each quadrant of ODE patients were significantly thickened than those of controls,and the superior cpRNFLT was the thickest,followed by the inferior,then the nasal and temporal cpRNFLT,and the difference was statistically significant (P ＜0.001).The mean cpTRT and cpTRT in each quadrant of ODE patients were significantly thickened than those of controls,and the inferior cpRNFLT was the thickest,followed by the superior,then the nasal and temporal cpRNFLT,and the difference was statistically significant (P ＜0.001).Conclusion Significantly thickening of cpRNFLT and cpTRT has been observed in patients with ODE as compared with controls,so both circumpapillary thickness parameters measured by OCT can be considered as a new non-invasive quantitative index for the detection and research of ODE.
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In vitro method as indispensable biological research and testing tools, has been widely used in cosmetics compliance test, biological mechanism research and functional materials screening. These methods include alternative methods which have been accepted by reguations and standardized test guide, non-testing method used for risk assessment and prediction, and a lot more diversification and individualization of in vitro methods. With the consensus formation of the whole industry and increase of application experience, innovation and optimization in vitro method will be constantly emerge. These technologies will be beneficial to improve competitiveness and to develop products to meet the needs of consumers.
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In vitro method as indispensable biological research and testing tools, has been widely used in cosmetics compliance test, biological mechanism research and functional materials screening. These methods include alternative methods which have been accepted by reguations and standardized test guide, non-testing method used for risk assessment and prediction, and a lot more diversification and individualization of in vitro methods. With the consensus formation of the whole industry and increase of application experience, innovation and optimization in vitro method will be constantly emerge. These technologies will be beneficial to improve competitiveness and to develop products to meet the needs of consumers.
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<p><b>BACKGROUND</b>Labyrinthine fistula (LF) is a very common clinical complication mainly caused by middle ear cholesteatoma. Whether the presence of different degree LF caused by middle ear cholesteatoma aggravates neurosensory hearing loss (NSHL) and what is the degree of the hearing loss caused by LF were still under controversial. This study aimed to investigate whether the LF degree is correlative with the age distribution, disease duration and hearing loss degree for cholesteatomatous patients.</p><p><b>METHODS</b>The files of 143 patients with middle ear cholesteatoma were selected and reviewed in a retrospective study. Seventy-eight patients with LF were divided into three types according to the degree of destruction of labyrinth. Sixty-five patients without LF were randomly chosen for control. Then, we compared the clinical characteristics of patients with or without labyrinthine fistulae caused by middle ear cholesteatoma.</p><p><b>RESULTS</b>According to the study, cholesteatomatous patients with LF were older and suffered longer disease duration than those without LF. Hearing loss is severe with high frequencies both in patients with and without LF. Moreover, inner ear impairment is correlative with the degree of destruction in labyrinth, and more severe destruction in labyrinth follow the more severe symptoms correlative with inner ear impairment.</p><p><b>CONCLUSION</b>Surgical intervention should be performed as early as possible for these cholesteatomatous patients.</p>
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Adolescent , Adult , Aged , Humans , Middle Aged , Cholesteatoma, Middle Ear , General Surgery , Fistula , Labyrinth Diseases , Retrospective StudiesABSTRACT
<p><b>OBJECTIVE</b>To explore the association of pathologic factors with the staging of metastatic lymph node ratio (rN) and metastatic lymph node number (pN), and to provide evidence for reasonable tumor staging in advanced gastric carcinoma (AGC).</p><p><b>METHODS</b>The clinicopathological data of 555 patients, who received radical resection for primary tumor of AGC between November 2003 and December 2011 in The First Affiliated Hospital of Xinjiang Medical University, were reviewed retrospectively. The clinicopathological factors influencing rN and pN were analyzed.</p><p><b>RESULTS</b>Univariate analysis showed that differentiation degree, vascular invasion, tumor diameter, gross type and invasion depth were significantly associated with rN or pN (all P<0.05). Histological type was significantly associated with rN (P<0.05), but not with pN. Logistic regression analysis revealed that vascular invasion, tumor diameter≥4 cm and invasion depth were independent risk factors for lymph node distant metastasis in AGC (all P<0.05). ROC curves showed that rN was consistent with pN in evaluating the diagnostic value of lymph node distant metastasis for tumor staging in AGC (P>0.05).</p><p><b>CONCLUSIONS</b>Vascular invasion tumor diameter≥4 cm and invasion depth are independent risk factors for lymph node metastasis in AGC based on either metastatic lymph node ratio (rN) or metastatic lymph node number (pN). The rN staging is consistent with the pN staging in evaluating the diagnostic value of metastatic lymph node for tumor staging in AGC.</p>
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Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Lymph Nodes , Pathology , Lymphatic Metastasis , Neoplasm Staging , Retrospective Studies , Stomach Neoplasms , PathologyABSTRACT
Research on molecular mechanisms of carcinogenesis plays an important role in diagnosing and treating gastric cancer. Metabolic profiling may offer the opportunity to understand the molecular mechanism of carcinogenesis and help to non-invasively identify the potential biomarkers for the early diagnosis of human gastric cancer. The aims of this study were to explore the underlying metabolic mechanisms of gastric cancer and to identify biomarkers associated with morbidity. Gas chromatography/mass spectrometry (GC/MS) was used to analyze the serum metabolites of 30 Chinese gastric cancer patients and 30 healthy controls. Diagnostic models for gastric cancer were constructed using orthogonal partial least squares discriminant analysis (OPLS-DA). Acquired metabolomic data were analyzed by the nonparametric Wilcoxon test to find serum metabolic biomarkers for gastric cancer. The OPLS-DA model showed adequate discrimination between cancer and non-cancer cohorts while the model failed to discriminate different pathological stages (I-IV) of gastric cancer patients. A total of 44 endogenous metabolites such as amino acids, organic acids, carbohydrates, fatty acids, and steroids were detected, of which 18 differential metabolites were identified with significant differences. A total of 13 variables were obtained for their greatest contribution in the discriminating OPLS-DA model [variable importance in the projection (VIP) value >1.0], among which 11 metabolites were identified using both VIP values (VIP >1) and the Wilcoxon test. These metabolites potentially revealed perturbations of glycolysis and of amino acid, fatty acid, cholesterol, and nucleotide metabolism of gastric cancer patients. These results suggest that gastric cancer serum metabolic profiling has great potential in detecting this disease and helping to understand its metabolic mechanisms.
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Adult , Aged , Aged, 80 and over , Female , Humans , Middle Aged , Metabolome , Stomach Neoplasms/blood , Biomarkers, Tumor/blood , Adenocarcinoma , Case-Control Studies , Gas Chromatography-Mass Spectrometry , Neoplasm Staging , Stomach Neoplasms/diagnosis , Stomach Neoplasms/pathologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of Evn-50 extracted from Vitex negundo on human breast cancer cell line MCF-7 and MCF-7/TAM-R cells in vitro.</p><p><b>METHODS</b>MCF-7 and tamoxifen-resistant MCF-7/TAM-R cells were treated with Evn-50,tamoxifen or combination of Evn-50 and tamoxifen. Cell proliferation inhibition rates were determined by MTT assay. The apoptosis rate and the change of cell cycle were detected by PI staining flow cytometry. Protein expression of phospho-MAPK 44/42 (Thr202/Tyr204),MAPK P44/42, phospho-AKT (Ser473) and AKT were detected with Western blotting.</p><p><b>RESULTS</b>The viability of MCF-7 cells was decreased in combination group [(28.65 ±11.43)%] and Evn-50 group [(53.02 ±15.14)%] compared with TAM group (P<0.01). The cell viability of MCF-7/TAM-R in combination group [(42.11 ±14.30)%] was significantly lower than that in TAM group [(92.18 ±13.16)%] (P<0.01). The cell apoptosis rate was dependent on the time of treatment in all groups,the effects on apoptosis and G2/M phase cells were most prominent at 72 h (P<0.01). Western blotting revealed that protein levels of phosphorylated AKT and p-MAPK44/42 decreased,while the expression of total AKT and MAPK44/42 was stable. In MCF-7/TAM-R cells,the expression of phosphorylation of AKT and MAPK44/42 protein was not changed in Evn-50 or TAM alone group,but significantly inhibited in the combination group at 72 h.</p><p><b>CONCLUSION</b>Evn-50 can inhibit cell growth and induce apoptosis in MCF-7 and MCF-7/TAM-R cells,it can reverse tamoxifen-resistance of MCF-7/TAM-R cells.The mechanisms may be related to the down-regulation of phosphorylated ERK1/2 in MAPK signal pathway and phosphorylated AKT in AKT signal pathway.</p>
Subject(s)
Female , Humans , Apoptosis , Breast Neoplasms , Drug Therapy , Metabolism , Pathology , Cell Cycle , Cell Proliferation , Drug Resistance, Neoplasm , Drugs, Chinese Herbal , Pharmacology , MCF-7 Cells , Phosphorylation , Proto-Oncogene Proteins c-akt , Metabolism , Signal Transduction , Tamoxifen , Therapeutic Uses , Vitex , ChemistryABSTRACT
<p><b>OBJECTIVE</b>To investigate the risk factors on female breast cancer in Zhejiang province.</p><p><b>METHODS</b>A case-control study was conducted in 200 cases of female breast cancer with histopathological diagnosis and 200 matched controls from Zhejiang province.</p><p><b>RESULTS</b>Univariate conditional logistic regression showed that family history of malignant tumor and breast cancer, housing decoration in last 10 years, mammary hyperplasia, adverse life events, bra with steel rings, sleeping with bra, high fat and pickle intake, poor sleep were positively related to breast cancer; whereas environmental friendly decoration materials, long decoration time interval, workplace condition, more lactation and parity, high fruits intake, sufficient sleep were negatively related to breast cancer. Multivariate conditional logistic regression analysis showed that the risk factors included family history of other tumors [odds ratio (OR)= 1.571,95% confidence interval(CI):1.029-2.396],mammary hyperplasia (OR=3.066,95%CI:1.834-5.126), job-related life events (OR=4.575,95%CI:1.690-12.390),the death of a loved one (OR=2.555,95%CI:1.475-4.424), wearing bra at night (OR=1.902,95%CI:1.177-3.072),high fat intake (OR=2.709,95%CI:1.546-4.749) and salted food (OR=2.460,95%CI:1.300-4.653). Factors as environmental friendly decoration materials (OR=0.517,95%CI:0.339-0.789),workplaces condition (OR=0.430,95%CI:0.243-0.762),more lactation (OR=0.109,95%CI:0.013-0.896),enough sleep (OR=0.424,95%CI:0.205-0.880) were protective factors.</p><p><b>CONCLUSION</b>Hereditary,psychological factors,lifestyle,environment and diet related factors are significantly associated with risk of breast cancer.</p>
Subject(s)
Female , Humans , Breast Neoplasms , Case-Control Studies , China , Epidemiology , Logistic Models , Multivariate Analysis , Risk FactorsABSTRACT
<p><b>BACKGROUND</b>This is a prospective clinical study based on a large sample gathered from multiple centers in China, subordinating to 10th Five-Year Plan of National Science & Technology Progression. We analyzed the high-risk factors inducing hypertensive disorders in pregnancy (HDP) and estimated the potential effect of anti-oxidants administration, including vitamin C (VC), vitamin E (VE) and Salvia Miltiorrhiza L (SML), a Chinese herb medicine, in amelioration of the high-risk factors in pregnancy.</p><p><b>METHODS</b>From April 2005 to July 2006, 4814 pregnant women from 24 national wide cooperative hospitals were involved in this prospective research. The participants were randomly divided into two groups: 1607 cases were in anti-oxidants group with administration of vitamins and SML; 3207 cases were in control group without any medicine given. Every participant was under monitoring for the morbidity of HDP and the high-risk factors were investigated in HDP cases in each group.</p><p><b>RESULTS</b>(1) The morbidity of HDP was 3.55% in anti-oxidants group vs. 4.18% in control group. No statistical difference existed between the two groups (P > 0.05). (2) In anti-oxidants group, the HDP morbidities among three subgroups: VC + VE + SML, VC + VE and SML only, were 5.51%, 3.05% and 5% respectively. It showed no statistical difference among three remedies (P > 0.05). (3) The related index of factors affecting HDP showed in intensity sequence as follows: family HDP history > profession > education level > age > body weight. The incidence of HDP in normal population was 3.51%, and the incidence of HDP in high-risk pregnant women (family HDP history, heavy physical labor, low education level (middle school and below), age ≥ 40, body mass index ≥ 24) was 5.84%, which was obviously higher than that in normal population (P < 0.01). In anti-oxidants group, the probability of HDP in women with high-risk factors was 3.81%, which was obviously lower than that in control group with high-risk factors at 7.14% (P < 0.01). (4) In control group, the morbidity of HDP in women with family HDP history (especially with sisters'), heavy physical labor, middle school and below, age ≥ 35 was: 50.00%, 15.22%, 6.33%, 26.28% and 5.75%, respectively, and that in anti-oxidants group was 0, 7.69%, 3.74%, 9.27% and 2.67%, respectively, which was obviously lower than that in control group.</p><p><b>CONCLUSIONS</b>The high-risk factors prone to induce HDP included: family history of HDP, heavy physical labor, low education level, aging and obesity. No impressive effect of anti-oxidants application was found in preventing HDP in general population but the remedy demonstrated positive effect on preventing HDP in pregnant women with high-risk factors.</p>
Subject(s)
Adolescent , Adult , Female , Humans , Pregnancy , Young Adult , Age Factors , Antioxidants , Therapeutic Uses , Body Weight , Hypertension , Drug Therapy , Logistic Models , Pregnancy Complications , Drug Therapy , Prospective Studies , Risk FactorsABSTRACT
<p><b>OBJECTIVE</b>To report nine cases of descending necrotizing mediastinitis (DNM) and to summarize the management experience.</p><p><b>METHODS</b>Between December 2005 and December 2008, nine patients (mean age, 55.7 years; age range, 38 to 78 years) with DNM were treated. Eight patients underwent surgical drainage of the involved cervical region and mediastinum (4 with cervical drainage alone; 4 with cervical drainage and right thoracotomy).</p><p><b>RESULTS</b>Two patients died, one of them refused surgical therapy and the other one died of multiorgan failure related to postoperative septic shock. Seven patients recovered. The mortality rate was 22%.</p><p><b>CONCLUSIONS</b>Delayed diagnosis and inadequate drainage are the main causes of high mortality rate of DNM. Aggressive surgical drainage and debridement of the neck and mediastinum by a multidisciplinary team of surgeons are very important in the treatment of DNM.</p>
Subject(s)
Adult , Aged , Humans , Male , Middle Aged , Focal Infection , MediastinitisABSTRACT
<p><b>OBJECTIVE</b>To investigate the methods and conditions for the isolation, purification and culture of human spermatogonial stem cells (SSCs) on the feeder layer cells of human embryonic fibroblasts (hEFs).</p><p><b>METHODS</b>SSCs isolated and purified from normal human fetal testicular tissues by sequential two-step enzyme digestion and Percoll uncontinuous density gradient centrifugation were cultured on the feeder layer cells of hEFs isolated from 5-9 weeks old human embryos. The surface markers SSEA-1 and OCT4 of the SSCs were detected by immunohistochemistry; the alkaline phosphatase (AKP) activity of the SSC clones measured; and the expressions of the SSC-related genes determined by RT-PCR.</p><p><b>RESULTS</b>SSCs survived, proliferated and formed colonies on the feeder layers, and the colonies were highly positive for SSEA-1 and OCT4, with strong AKP activity and high expressions of the SSC-related genes.</p><p><b>CONCLUSION</b>The feeder layer of hEFs supports the growth of human spermatogonial stem cells.</p>