Human hair follicle-derived mesenchymal stem cells: Isolation, expansion, and differentiation.

Hair follicles are easily accessible skin appendages that protect against cold and potential injuries. Hair follicles contain various pools of stem cells, such as epithelial, melanocyte, and mesenchymal stem cells (MSCs) that continuously self-renew, differentiate, regulate hair growth, and maintain skin homeostasis. Recently, MSCs derived from the dermal papilla or dermal sheath of the human hair follicle have received attention because of their accessibility and broad differentiation potential. In this review, we describe the applications of human hair follicle-derived MSCs (hHF-MSCs) in tissue engineering and regenerative medicine. We have described protocols for isolating hHF-MSCs from human hair follicles and their culture condition in detail. We also summarize strategies for maintaining hHF-MSCs in a highly proliferative but undifferentiated state after repeated in vitro passages, including supplementation of growth factors, 3D suspension culture technology, and 3D aggregates of MSCs. In addition, we report the potential of hHF-MSCs in obtaining induced smooth muscle cells and tissue-engineered blood vessels, regenerated hair follicles, induced red blood cells, and induced pluripotent stem cells. In summary, the abundance, convenient accessibility, and broad differentiation potential make hHF-MSCs an ideal seed cell source of regenerative medical and cell therapy.

Heavy metals in wild marine fish from South China Sea: levels, tissue- and species-specific accumulation and potential risk to humans.

Heavy metal pollution in marine fish has become an important worldwide concern, not only because of the threat to fish in general, but also due to human health risks associated with fish consumption. To investigate the occurrence of heavy metals in marine fish species from the South China Sea, 14 fish species were collected along the coastline of Hainan China during the spring of 2012 and examined for species- and tissue-specific accumulation. The median concentrations of Cd, Cr, Cu, Zn, Pb and As in muscle tissue of the examined fish species were not detectable (ND), 2.02, 0.24, 2.64, 0.025, and 1.13 mg kg(-1) wet weight, respectively. Levels of Cu, Zn, Cd and Cr were found to be higher in the liver and gills than in muscle, while Pb was preferentially accumulated in the gills. Differing from other heavy metals, As did not exhibit tissue-specific accumulation. Inter-species differences of heavy metal accumulation were attributed to the different habitat and diet characteristics of marine fish. Human dietary exposure assessment suggested that the amounts of both Cr and As in marine wild fish collected from the sites around Hainan, China were not compliant with the safety standard of less than 79.2 g d(-1) for wild marine fish set by the Joint FAO/WHO Expert Committee on Food Additives. Further research to identify the explicit sources of Cr and As in marine fish from South China Sea should be established.

[Effects of linggui zhugan decoction combined calorie restriction on the insulin resistance of model rats and mechanisms research].

OBJECTIVE: To explore the effects of Linggui Zhugan Decoction (LZD) combined calorie restriction on fasting plasma glucose (FPG), the insulin resistance (IR), and the peroxisome proliferator-activated receptor gamma (PPAR-gamma) of IR model rats. METHODS: Totally 48 male Wistar rats were randomly divided into the control group, the model group, the calorie restriction group, and the TCM + calorie restriction group, 12 in each group. Ordinary forage was given to those in the control group, and high fat diet was fed to those in the rest 3 groups for 12 weeks to establish the IR model. After successful modeling, rats in the control group and the model group were continually fed with the original farage for 4 days. The normal saline at the daily dose of 20 mL/kg was given to them by gastrogavage. The normal saline at the daily dose of 20 mL/kg was given to rats in the calorie restriction group by gastrogavage after 4-day calorie restriction. LZD at the daily dose of 20 mL/kg was given to rats in the TCM +calorie restriction group by gastrogavage after 4-day calorie restriction. The body weight, FPG, serum fasting insulin (FINS), insulin resistance index (IRI), and the protein expression of PPAR-y in the omental adipose tissue were compared. RESULTS: After 4-day calorie restriction, the body weight obviously decreased in the calorie restriction group and the TCM +calorie restriction group, when compared with the model group (P <0.01). There was no statistical difference between the former two groups (P >0.05). The FINS and IRI obviously decreased in the calorie restriction group (P <0.01, P <0.05). The FPG, FINS, and IRI significantly decreased in the TCM + calorie restriction group (P <0. 05, P <0.01). The protein expression of PPAR-gamma obviously decreased in the calorie restriction group and the TCM + calorie restriction group (P <0.01).The phlegm dampness state was alleviated, with more significant effects shown in the TCM + calorie restriction group. CONCLUSIONS: LZD combined calorie restriction could reduce the body weight, FPG, and IRI of IR rats. Besides, it showed better effects than calorie restriction alone. Its effects in improving IR might be correlated with inhibiting the activities of PPAR-gamma. Meanwhile, it might play a role in inhibiting the differentiation of fat cells.

Acrolein induces Alzheimer's disease-like pathologies in vitro and in vivo.

The pathologic mechanisms of Alzheimer's disease (AD) have not been fully uncovered. Acrolein, a ubiquitous dietary pollutant and by-product of oxidative stress, can induce cytotoxicity in neurons, which might play an important role in the etiology of AD. Here, we examined the effects of Acrolein on the AD pathologies in vitro and in vivo. We found Acrolein induced HT22 cells death in concentration- and time-dependent manners. Interestingly, Acrolein increased proteins' levels of amyloid precursor protein (APP), ß-secretase (BACE-1) and the amyloid ß-peptide transporter receptor for advanced glycation end products, and decreased A-disintegrin and metalloprotease (ADAM) 10 levels. In vivo, chronic oral exposure to Acrolein (2.5 mg/kg/day by intragastric gavage for 8 weeks) induced mild cognitive declination and pyknosis/atrophy of hippocampal neurons. The activity of superoxide dismutase was down-regulated while the level of malondialdehyde was up-regulated in rat brain. Moreover, Acrolein resulted in activation of astrocytes, up-regulation of BACE-1 in cortex and down-regulation of ADAM-10 in hippocampus and cortex. Taken together, our findings suggest that exposure to Acrolein induces AD-like pathology in vitro and in vivo. Scavenging Acrolein might be beneficial for the therapy of AD.

[A study on apoptosis and apoptotic mechanisms of HL-7702 cell line induced by methylmercury].

OBJECTIVE: To study the apoptotic effect and mechanisms of methylmercury (MeHg) on HL-7702 cell line in vitro. METHODS: In this study, the cell apoptosis was observed by AO/EB method and FCM method; the mitochondrial membrane potential was detected by FCM; and the expression of proteins related to apoptosis was measured by immunocytochemical method. RESULTS: After exposure to MeHg for 24 h in different doses, apoptotic rate ascended with the increasing of MeHg concentration. By AO/EB method, cell apoptotic ratio of negative control group was (2.62 +/- 0.19)%, cell apoptotic ratio of 10-50 micromol/L exposure groups were (7.97 +/- 0.64)%, (12.66 +/- 0.76)%, (19.16 +/- 0.87)%, (18.42 +/- 0.88)%, and (11.52 +/- 0.63)%, there were significant differences between the exposure and negative control groups (q values were 17.057, 32.009, 52.732, 50.373, 28.375; P<0.05). Mitochondrial membrane potential descended with the increase of MeHg, mitochondrial membrane potential of negative control group was (10.23 +/- 3.43) mV, mitochondrial membrane potential of 10-50 micromol/L exposure groups were (3.25 +/- 0.66), (3.03 +/- 0.35), (1.68 +/- 1.26), (1.69 +/- 1.13) and (1.77 +/- 0.88) mV, and there was significant differences between exposure and negative control groups (q values were 9.569, 9.871, 11.722, 11.708, 11.598; P<0.05). The expression of Bax, Bcl-2, CytC, Caspase-3 and AIF enhanced with the increase of MeHg, Bax/Bcl-2 ratio also appeared a trend of increase. Bax expression integral optical density (IOD) of negative control group was (21295.86 +/- 1969.81), Bax expression IOD of 10, 20, 30 micromol/L groups were 42807.87 +/- 4416.64, 55651.65 +/- 4662.72, and 72708.56 +/- 910.10, there were significant differences in Bax expression between 10, 20, 30 micromol/L groups and negative control group (q values were 14.191, 14.320, 33.917; P<0.05); Bcl-2 expression IOD of negative control group was (12588.33 +/- 4091.02), Bcl-2 expression IOD of 10, 20, 30 micromol/L groups were 20539.16 +/- 4906.09, 23689.97 +/- 2281.42, and 28692.80 +/- 4655.86, there were significant differences in Bcl-2 expression between 10, 20, 30 micromol/L groups and negative control group (q values were 4.322, 6.035, 8.754; P<0.05); and AIF expression IOD of negative control group was (12942.72 +/- 457.94), AIF expression IOD of 10, 20, 30, 40 micromol/L groups were 16973.57 +/- 1922.87, 29998.91 +/- 6803.58, 52467.16 +/- 1916.25 and 106342.53 +/- 1273.19, there were significant differences in AIF expression between 20, 30 and 40 micromol/L groups and negative control group (q values were 11.449, 26.530, 62.692; P<0.05). CONCLUSION: MeHg could induce apoptosis on HL-7702 cell line in vitro. The mechanisms could be related to mitochondrial pathway in apoptosis.

[Effects of over-expressed Smac gene coupling with cisplatin on proliferation and apoptosis of hepatocarcinoma cells].

OBJECTIVE: To investigate the effects of over-expressed Smac gene combined with cisplatin (CDDP) on proliferation and apoptosis of hepatic carcinoma cells. METHODS: The recombinant plasmid pcDNA3.1+-hSmac was introduced into the human hepatic carcinoma SMMC-7721 cells using a liposome-mediated method. The expression of Smac protein was detected by Western blot and flow cytometry. The cells were treated with three different doses of CDDP, 5, 15 and 25microg/ml, for 24 hours after the transfection. MTT colorimetry was used to detect the cellular growth-inhibitory effects; acridine orange-ethidium bromide fluorescent staining (AO/EB) and flow cytometry with annexin V-PI double staining METHODS: were used to detect the changes of cell apoptosis. RESULTS: Western blot and flow cytometry results demonstrated that the Smac protein level in SMMC-7721 cells was significantly increased after the transfection (P less than 0.01). Compared with that of the control group, the over-expressed Smac gene inhibited the cell growth and induced cell apoptosis (P less than 0.01). After being treated with CDDP, the inhibitory rates were increased significantly with increasing concentrations of CDDP compared with that of the control group, and the inhibitory rate of the CDDP-treated plus Smac group was significantly higher than that of the CDDP-treated group (P less than 0.01). The results detected by AO/EB and flow cytometry demonstrated that the apoptotic rates of CDDP-treated plus Smac group were higher than those of the CDDP-treated group (P less than 0.01). The results demonstrated that the Smac over-expression enhanced the effects of cell growth inhibition and apoptotic promotion induced by CDDP. CONCLUSION: The pro-apoptotic Smac gene could be over-expressed in hepatocarcinoma SMMC-7721 cells and inhibit cell growth and induce apoptosis. Moreover the over-expressed Smac could enhance the chemotherapeutic sensitivity of SMMC-7721 to cisplatin. This experimental work may help in further study on the regulatory mechanism of Smac in apoptosis and improve the chemotherapeutic effect on hepatoma.

[Effect of capsule yindanxinnaotong (YD) on blood hemorheology and the score of blood stasis in patients with MS complicated ACS].

OBJECTIVE: To observe the effect of Capsule YD on blood hemorheology and the score of blood stasis in patients with MS complicated ACS, and to investigate the clinical effect of Chinese medicine of promoting blood flow on those diseases. METHODS: 63 cases were divided into two groups. Control group was treated with routine methods and YD group with additional YD. The course of treatment was 3 months. Blood hemorheology and the score of blood stasis were observed. RESULTS: Blood viscosity high shear rate, fibrinogen, the index and deformed index of Erythrocyte aggregation before and after treatment in both groups were improved (P < 0.05). And the index of Erythrocyte aggregation in YD group was significantly improved comparing to that in control group (P < 0.05). The symtom of stethalgia, frequency of episode, and blood stasis on tongue, lip and gums were also improved in YD group. The value of blood stasis in YD group was significantly decreased comparing to that in control group. CONCLUSIONS: Capsule YD can improve the angina pectoris and decrease the value of blood stasis in patients with ACS, and also improve their hemorheology. We should presume the Chinese medicine of activating blood circulation to dissipate blood stasis improve the metabolism and treat angina pectoris by improving blood hyperviscosity.

DNA damage of germ cell of rat induced by nitrotoluene chemicals.

2,4-dinitrotoluene(2,4-DNT), 2,6-dinitrotoluene(2,6-DNT) and 4-nitrotoluene(4-NT) are typical pollutants in the Songhua River of Northeast China. Sertoli/germ cell cocultures and single cell gel electrophoresis(SCGE) are applied to investigate whether they have genotoxicity on DNA damage of germ cell of Kunming male rat. The results showed that all three nitrotoluene compounds tested could induce DNA single-strand breaks of the germ cell. A significant relationship is found between logarithm dose and the degree of DNA damage, which implies that 2,4-DNT, 2,6-DNT and 4-NT have genotoxicity and can induce the germ cell DNA strand to break in vitro.

[Clinical and molecular-biological study of a May-Hegglin anomaly family].

OBJECTIVE: To study the changes of platelet in May-Hegglin anomaly (MHA) and the molecular pathogenesis mechanism. METHODS: Peripheral blood was drawn from the MHA proband, her father and her uncle. Platelet count and morphology were examined by automatic blood cell counter and microscopy, respectively. The platelet membrane protein was examined by flow cytometry. Membrane antibodies were determined by ELISA. PCR was used to amplify the exons 25, 31 approximately 32, 38 and 40 of the MYH 9 gene in the MHA patient and her diseased father. Furthermore, PCR products were sequenced, a specific point mutation was identified and inclusions (Dohle's body) in the neutrophil was detected by indirect immunofluorescence technique. RESULTS: It was proved that in MHA patients, platelet count was higher by cell counter than by microscope (P < 0.01). Giant platelet was 94% but platelet membrane proteins (CD41, CD61, CD42A, CD42b) were in normal range. Membrane antibodies was undetectable. An A5521G mutation (GAG-->AAG) in the exon 38 was found in the proband and her diseased father, resulting in a characteristic change of NMMHC-A1841 (Glutamic acid-->Arginine), which was not found in other members of the family and in normal controls. Spindle-like inclusions with fluorescence were clearly displayed in neutrophil cytoplasm. CONCLUSION: The molecular pathogenesis mechanism of May-Hegglin anomaly is the mutation in MYH 9 gene.

[Mid- to long-term effects of delayed percutaneous coronary intervention on left ventricular function and prognosis in patients with acute myocardial infarction].

OBJECTIVE: To investigate the mid- to long-term effects of delayed percutaneous coronary intervention (PCI) on the left ventricular function and clinical outcome of patients with acute myocardial infarction (AMI). METHODS: PCI (including percutaneous transluminal coronary angioplasty and stenting) was performed in 42 patients within 1 to 2 weeks following the onset of AMI (PCI group), with another 31 patients who were admitted within the same period to receive medication for AMI serving as the control group. The patients in both groups were observed for comparison of the occurrence of reinfarction and angina, mortality at 1 and 6 months, and findings by ultrasound cardiograms (UCG). RESULTS: In PCI group, the left ventricular function were obviously improved as compared with the control group (P<0.01) 1 month after the onset of AMI, showing greater improvement at 6 months (P<0.01). No death or reinfarction occurred in the PCI group, with only 1 patients experiencing angina 5 months after PCI. In control group, death occurred in 2 cases, reinfarction in 1 case, recurrent angina in 4 cases (include 2 cases of early postinfarction angina). CONCLUSION: Delayed PCI may significantly improve the prognosis of patients with AMI and prolong their survival without cardiovascular accidents and ameliorate their left ventricular functions, with high success rate of the operation.