BACKGROUND:Separating umbilical cord blood stem cells using tubes has low efficiency,and microbial contamination easily occurs during this process,therefore,safety cannot be ensured in clinical application.lt is urgent to find a method for separating umbilical cord blood stem cells to treat femoral head necrosis.OBJECTIVE:To establish a high efficient,safe,and clinically valuable method to separate umbilical cord blood stem cells.DESIGN,TIME AND SETTING:A self-control experiment was performed at the First Department of Surgery,Zhengzhou Second People's Hospital,Institute of Blood Constituent Application,Henan Red Cross Blood Centre between February 2006 and August 2007.PARTICIPANTS:Eight male patients with femoral head necrosis,averaging 40.6 years of age,were included in this study.Of these patients,4 had the history of hormone application.An average of 90 mL umbilical cord blood was harvested from each healthy normal full term neonate from Maternal and Children Health Care Hospital of Zhengzhou City.The quadruple bags used for separating umbilical cord blood stem cells consisted of 1 main bag,1 empty bag,and 2 physiological saline bags,provided by Shandong Weigao Holding,China.METHODS:Within 6 hours after collection,umbilical cord blood was centrifuged in the empty bag of quadruple bag,which was connected with an aseptic filling machine.After centrifugation,partial blood plasma was discarded,and the remaining erythrocytes were thoroughly mixed by adding hetastarch.Five minutes later,the mixture was diluted with physical saline at 1:1.Umbilical cord blood was slowly added into the main bag (at 1:1),in which,human lymphocyte separating medium was pre-added.After cantrifugation,the upper layer of solution,i.e.,monocyte-rich solution,was transferred into another empty bag.Within 24hours of preservation,after suspension with umbilical blood plasma,umbilical cord monocytes were transfused into patients with femoral head necrosis via superficial vein on the hand back,monocytes≥1×108/portion,2 portions once.There were three treatment courses,each involving three transfusion sessions,one session every 4 days,and a 2-3-month interval between two treatment courses.MAIN OUTCOME MEASURES:Cell recovery rate and cell viability of umbilical cord blood monocytes and improvements in clinical symptoms.RESULTS:The separation of quadruple bags could obtain umbilical cord blood monocytes with high recovery rate.Furthermore,microbial contamination hardly occurred in the process of separation.Hip joint pain relieved or disappeared to different extents in all 8 patients,with an effective rate of 100%.Abduction and internal rotation of hip joint,ambulation distance,and gait were markedly improved.At 6 months after cell transplantation,5 patients presented with changed bone density in femoral head necrosis regions,2 showed normal femoral head morphology,and the remaining 1 exhibited no obvious changes.Joint effusion was reduced or disappeared in 12 hips.Magnetic resonance images showed that femoral head morphology had been improved in various degrees in 9 hips,but no changes in 3 hips.No complications,fever,or allergies occurred during and after cell transplantation.CONCLUSION:The method of separating stem cells from umbilical cord blood in junction with aseptic interface technology is highly effective,safe,and clinically valuable.Multiple intravenous transfusions of umbilical cord blood stem cells provide a novel approach for systemic treatment of femoral head necrosis.

Humans ; Siblings ; Forensic Medicine ; Forensic Genetics

OBJECTIVE: To study the level of circulating Alarin in obese children and its association with various metabolic parameters. METHODS: A total of 86 obese children with a body mass index (BMI) above the 95th percentile were enrolled as the obesity group, and 82 healthy children, matched for age and sex, with a BMI below the 85th percentile were enrolled as the healthy control group. According to the presence or absence of insulin resistance (IR), the obesity group was further divided into an IR group with 27 children and a non-IR group with 59 children. Related anthropometric parameters, including body height, body weight, systolic blood pressure (SBP), and diastolic blood pressure (DBP), were measured, and BMI was calculated. Total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), uric acid (UA), fasting insulin (FINS), and fasting blood glucose (FBG) were measured. The area under the receiver operating characteristic curve (AUC) for glucose and insulin, Homeostasis Model Assessment of Insulin Resistance (HOMA-IR), and whole-body insulin sensitivity index (WBISI) were calculated. ELISA was used to measure the level of circulating Alarin. RESULTS: The obesity group had a significantly higher level of circulating Alarin than the healthy control group (P<0.01). The IR group had a significantly higher level of circulating Alarin than the non-IR group (P<0.01). Circulating Alarin was positively correlated with BMI, TG, FBG, AUC-glucose, AUC-FINS, and HOMA-IR (P<0.05) and was negatively correlated with WBISI (P<0.05). The circulating Alarin level had a linear regression relationship with BMI, FBG, and HOMA-IR, among which HOMA-IR had the greatest influence on the circulating Alarin level (P<0.05). CONCLUSIONS There is a significant increase in the circulating Alarin level in obese children, which may be associated with the development of obesity and IR.
Blood Glucose ; Body Mass Index ; Child ; Galanin-Like Peptide ; Humans ; Insulin ; Insulin Resistance ; Obesity

Human Interleukin-11(hIL-11)has no Cys residue in its natural form.By site-directed mutagenesis,a Cys residue can be introduced to replace the 1st residue Gly and the rhIL-11 was chemically modified by using 20 kDa mPEG-maleimide conjugated to this site.The mPEG-hIL-11 conjugate was purified and showed a single band on SDS-PAGE with an apparent molecular weight.The biological activity of purified mPEG-hIL-11 was determined using a dependent cell line 7TD1.The remaining biological activity of PEGylated-rhIL-11 was 30% of native rhIL-11,suggesting chemical modification of rhIL-11 by PEG is a promising approach for improving the pharmacological efficacy.

OBJECTIVETo investigate the expressions of PPAR-gamma/PGC-1alpha and Nrf2/gamma-GCS-h in lung of guinea pigs with bronchial asthma, and to explore the roles of them.

METHODSForty adult male guinea pigs were randomly divided into 4 groups: the control group (group A), asthmatic group ( group B), dexamethasone group (group C) and rogridone group (group D), 10 guinea pigs in each group. The asthmatic model was established by the ovalbumin challenge method. Expressions of PPAR-gamma/PGC-1alpha and Nrf2/gamma-GCS-h mRNA in lung tissue were assayed by in situ hybridization. Expressions of PPAR-gamma/PGC-1alpha and Nrf2/gamma-GCS-h protein were detected by immunohischemistry and by Western blot.

RESULTSIn situ hybridization showed that the expressions of PPAR-gamma/PGC-1alpha and Nrf2/gamma-GCS-h mRNA in lung tissue were the lowest in group B and the comparison among groups showed statistical significant (all P < 0.01). Immunohistochemistry and Western blot indicated that the value of PPAR-gamma/PGC-1alpha and Nrf2/gamma-GCS-h protein in lung tissue were the lowest in group B, and expressed primarily in nucleus, the differences being statistically significant (all P < 0.01). There was positive correlation between PPAR-gamma and PGC-1. gamma-GCS-h mRNA also positively correlated between PPAR-gamma/PGC-1alpha and Nrf2 in nucleus, and the expression of Nrf2 was also positively correlated with PPAR-gamma/ PGC-1alpha.

CONCLUSIONIn acute asthmatic models induced by ovalbumin, the expressions of PPAR-alpha/PGC-1alpha and Nrf2/gamma-GCS-h were decreased, and PPARgamma/PGC-1alpha could up-regulate the expressions of Nrf2/gamma-GCS-h to increase the antioxidant defense of tissues, thus being implicated that PPARgamma/PGC-1alpha might play important roles in the pathogenesis and prevention of asthma.

Animals ; Asthma ; chemically induced ; physiopathology ; Glutamate-Cysteine Ligase ; genetics ; metabolism ; Guinea Pigs ; Lung ; metabolism ; Male ; NF-E2-Related Factor 2 ; genetics ; metabolism ; Ovalbumin ; PPAR gamma ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Transcription Factors ; genetics ; metabolism

OBJECTIVE: To explore the obesity distribution in old people and the relation between senile obesity and health. METHODS: First, a questionnaire was designed which included chronic disease history, body mass index (BMI), physiological value, biochemistry index, anti-oxidation index, diagnosis of diseases, etc. Second, the measure and detection methods were unified; and the last, the investigation was made along with daily clinical work by clinicians. RESULTS: We received 391 questionnaires. The overweight rate was 36.1% and the obesity rate was 7.9% . Total anti-oxidation activity in serum (TAS) and superoxide dismutase (SOD) decreased with body mass index (BMI), and the value in the obesity group was the lowest; Malonaldehyde (MDA) of overweight obesity was the largest. The mean blood pressure, blood fat, and blood glucose as well as the prevalence of cardiovascular disease, hyperlipemia, and glycuresis increased with BMI; and the value in the obesity group was the largest. CONCLUSION The prevalence of the senile obesity was below the average and the senile obesity complications were various and serious, and perhaps related to imbalance of free radical's production and cleanup, so the senile obesity seriously harmed old people's health.
Aged ; Body Mass Index ; Cardiovascular Diseases ; epidemiology ; etiology ; Cerebrovascular Disorders ; epidemiology ; etiology ; China ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Obesity ; complications ; epidemiology ; Overweight ; Prevalence ; Risk Factors

OBJECTIVETo explore the effects of percutaneous transhepatic radiofrequency ablation (PRFA) combined with tumor edge of percutaneous absolute ethanol injection (PEI) on liver cancer adjacent to major blood vessels.

METHODSSeventy five patients with liver cancer adjacent to major blood vessels were randomly divided into two groups: PRFA+PEI therapy group (38 cases) and PRFA control group (37 cases). Tumor necrosis rate, AFP levels, local recurrence rate, median for survival time and cum survival were used as the evaluation index to evaluate the efficacies of the two methods.

RESULTSTumor necrosis rates of the therapy group and the control group were 84.2% and 54.1% (P < 0.01), respectively; AFP levels of therapy group and control group at 1, 3, 6 and 12 months after treatment were (105.0 ± 35.5) μg/L, (28.4 ± 4.3) μg/L, (58.6 ± 6.7) μg/L, (89.5 ± 12.5) μg/L and (137.2 ± 34.6) μg/L, (84.2 ± 18.4) μg/L, (106.6 ± 20.3) μg/L, (173.7 ± 32.0) μg/L, respectively. The rates of therapy group was significantly lower than of control group. Local recurrence rates of the therapy group and control group were 2.6%, 7.9%, 13.2% and 31.6% vs 10.8%, 21.6% , 40.5% and 62.1% (P < 0.05) at 3, 6, 12 and 24 months after treatment, respectively. Median for survival time of the therapy group and control group were 28.0 ± 2.8 months and 19.0 ± 3.6 months, respectively. Cum survival of the therapy group and control group were 84.2%, 78.9%, 60.5% and 31.6% vs 78.4%, 67.6%, 37.8% and 8.1% (P < 0.05) at 6, 12, 24 and 36 months after treatment, respectively.

CONCLUSIONPEI as a supplementary treatment of PRFA can effectively improve the treatment of liver cancer adjacent to major blood vessels and significantly reduce the local recurrence rate and improve long-term survival rates.

Adult ; Aged ; Bile Duct Neoplasms ; Carcinoma, Hepatocellular ; pathology ; therapy ; Catheter Ablation ; Combined Modality Therapy ; Ethanol ; administration & dosage ; Female ; Humans ; Liver Neoplasms ; pathology ; therapy ; Male ; Middle Aged ; Retrospective Studies ; Survival Rate ; Treatment Outcome

OBJECTIVETo investigate whether the polymorphism of DNA repair genes XPC (Ala499Val and Lys939Gln) and XPG (His1104Asp) is associated with the susceptibility to hepatocellular carcinoma (HCC).

METHODSA hospital-based case-control study was conducted in 500 cases with HCC and 507 controls. Genotypes of XPC and XPG were determined by real-time polymerase chain reaction with the TaqMan MGB probe.

RESULTSCompared to the CC genotype, the CT genotype and the TT genotype of XPC Ala499Val were not associated with the susceptibility to HCC (adjusted OR = 1.34, 95% CI: 0.85-2.12; adjusted OR = 1.30, 95% CI: 0.68-2.51, respectively). Compared to the AA genotype, the AC genotype and the CC genotype of Lys939Gln were not associated with the susceptibility to HCC (adjusted OR = 1.20, 95% CI: 0.78-1.85; adjusted OR = 1.81, 95% CI: 0.88-3.73, respectively). Compared to the CC genotype, the CG genotype and the GG genotype of XPG His1104Asp were not associated with the susceptibility to HCC (adjusted OR = 0.85, 95% CI: 0.56-1.27; adjusted OR = 1.12, 95% CI: 0.67-1.87, respectively) However, the stratified analysis revealed that the females with the AC+CC genotype of XPC Lys939Gln had increased risk of HCC compared to those with AA genotype (OR = 2.17, 95% CI: 1.01-4.64).

CONCLUSIONOur results suggest that XPC and XPG polymorphisms do not independently affect on the susceptibility to HCC, but the joint effect of C allele of XPC Lys939Gln and female may modify the risk of HCC.

Carcinoma, Hepatocellular ; genetics ; Case-Control Studies ; DNA Repair ; DNA-Binding Proteins ; genetics ; Endonucleases ; genetics ; Female ; Genetic Predisposition to Disease ; Genotype ; Humans ; Liver Neoplasms ; genetics ; Male ; Middle Aged ; Nuclear Proteins ; genetics ; Polymorphism, Single Nucleotide ; Transcription Factors ; genetics

Objective To observe the protective mechanisms of zingiber officinalis extract on the lens of diabetic rats induced by streptozotocin (STZ).Methods Totally 60 SD rats were randomly divided into A group (normal control rats),B group (diabetic rats),C1 group (DM +50 mg · kg-1 ginger gavage),C2 group (DM + 100 mg ·kg-1 ginger gavage) and C3 group (DM + 300 mg · kg-1 ginger gavage).Each group had 12 rats.And rats in A group received intraperitoneal injection of the same volume of normal saline,while the other groups were intraperitoneally injected with 65 mg · kg-1streptozotocin (STZ).When the animal model was successfully established,both A group and B group were administered orally with normal saline,and the C1,C2 and C3 groups were administered orally with ginger rhizome extract.The changes in blood glucose and lens were observed every week.The rats were sacrificed in succession at 4 week,8 week,12 week,and the lens was removed immediately.The content of aldose reductase (AR) was detected by ELISA,and the expression of glycosylation end products (AGEs) was detected by Western blot.The superoxide dismutase (SOS) activity and malondialdehyde (MDA) content were also detected.Fluorescent Tunel staining was used to detect the apoptosis of the lens epithelial cells.And finally,scanning electron microscopy was used to observe the ultrastructural changes in the lens.Results The activity of SOD in the lens of diabetic rats showed a decreasing trend with statistical significance (all P ＜ 0.05).Changes in the content of MDA in the lens of rats in B,C1,C2 group were statistically significant in 4,8 and 12 weeks after successful modeling (all P ＜ 0.05),but no significant difference in A group and C3 group (both P ＞ 0.05).The persistent increase of AR in group B (P =0.003).The content of AR of the lens in C1,C2,C3 group showed a decreasing trend,and the differences were statistically significant (all P ＜ 0.05).There were significant differences in the expression of AGEs in C1,C2 and C3 group (all P ＜ 0.05).The degree of cortical fiber destruction in group B was progressively aggravated,but the degree of cortical destruction in C1,C2 and C3 group decreased with the increase of ginger gavage concentration through the scanning electron microscopy.It was observed that there was no significant difference in the apoptotic rate of LECs in A group (P =0.191),but the apoptosis of LECs in the rest group showed a rising trend,and the differences were statistically significant (all P ＜ 0.05).Conclusion The effects of ginger gavage extract can delay the opacification of lens and slow down the diabetic development in rats with a dose-independence manner.Ginger gavage extract may play a protective role on the lens of diabetic rats by inhibiting the activity of AR,oxidative stress and the production of AGEs as well as suppress the apoptosis of lens epithelial cells.

Objective To explore the effects of estradiol on the telomerase activity and apoptosis of lens epithelial cells and its mechanisms.Methods Fifty adult female Sprague-Dawley rats were randomly divided into five groups (n =10):ovariectomized group,estradiol group 1,estradiol group 2,estradiol group 3 and sham group.All rats were prepared for ovariectomized models except that of the sham group.After 2 weeks of the operation,rats in the 5 groups were given naphthalene solution through a stomach tube,and meanwhile,estradiol group 1,estradiol group 2,estradiol group 3 received estradiol valerate with the dose of 0.21 mg · kg-1 · d-1,0.42 mg · kg-1 · d-1 and 0.84 mg · kg-1 · d-1,accordingly,while rats in the ovariectomized group and sham group were given 9 g · L-1 Nacl by stomach perfusion with a dose of 0.42 mg · kg-1 ·d-1.Then,the opacity of the lens in each group was examined by a slit lamp microscope every week.After 12 weeks of naphthalene solution administration,all rats were sacririced and the serum estradiol concentration was determined by radioimmunoassay.Next,the lenses were taken out,and TERT mRNA expression of lens epithelial cells (LEC) was measured by RT-qPCR,and finally,the apoptotic rate was detected by TUNEL method.Results The opacity of lens in the ovariectomized group was different from that in the estradiol group 1,estradiol group 2,estradiol group 3 and sham group,with statistical significances (all P ＜ 0.05),and the opacity of lens in the estradiol group 1,2,3 and sham group were mild and occurred later.The serum estradiol concentration in the ovariectomized group (8.19 ± 1.45)ng · L-1 was significantly lower than that of estradiol groupl,2,3 and sham group,and there were significant differences (all P ＜ 0.05).Thc relative expression of TERT mRNA in LEC in the ovariectomized group (0.371 2-±0.056 4) was significantly lower than that in estradiol group 1,2,3,but the apoptotic rate of LEC (0.602 1 ±0.010 8) was obviously higher than that of estradiol group 1,2,3 in a dose-dependent manner,and there were significant differences (all P ＜ 0.05).Pearson correlation analysis showed the relative expression of LEC TERT mRNA in rats was negatively correlated with the apoptosis rate(r =-0.859,P ＜ 0.05).Conclusion Estradiol can up-regulate TERT mRNA expression and enhance telomerase activity of LEC in naphthalene-induced ovariectomized female rats in a dose-dependent manner.Estradiol can inhibit the LEC apoptosis in naphthalene-induced ovariectomized female rats,and the mechanism may be related to the increase of telomerase activity in the LEC.