BACKGROUND:Stem cel transplantation, in recent years, has become a preferred treatment for premature ovarian failure. Umbilical cord blood mononuclear cel s containing a large number of mesenchymal stem cel s, immature stem/progenitor cel s, and endothelial progenitor cel s can be used as an important source of stem cel transplantation.
OBJECTIVE:To study the therapeutic efficacy of human umbilical cord blood mononuclear cel transplantation on radioactive premature ovarian failure in nude mice.
METHODS:120 female BALB/C nude mice were randomly divided into four groups:blank control group without any intervention;model group, intravenous transplantation group, and in situ transplantation group exposed to 60Coγrays, 0.5 Gry per day, for 30 days. After 30 days, premature ovarian failure models were made in the latter three groups. Then, nude mice in the latter three groups were given bilateral ovary injection of 10 μL DMEM, tail vein injection of 10 μL human umbilical cord blood mononuclear cel s (1×1013/L), and bilateral ovary injection of 10 μL human umbilical cord blood mononuclear cel s (1×1013/L), respectively. Thirty days after cel transplantation, serum levels of estradiol, fol icle hormone, luteinizing hormone, inhibin B and vascular endothelial growth factor were detected, cel apoptosis in the ovary tissue and cel survival were observed pathological y.
RESULTS AND CONCLUSION:After modeling, the serum levels of estradiol, inhibin B and vascular endothelial growth factor were significantly reduced (P<0.01), while fol icle hormone and luteinizing hormone levels increased (P<0.01). After transplantation, these indexes were al improved in the in situ transplantation group (P<0.01), and reduced fol icle hormone and luteinizing hormone levels were visible in the intravenous transplantation group (P<0.05, P<0.01). Compared with the model group, in situ transplantation and intravenous injection of human umbilical cord blood mononuclear cel s could effectively reduce cel apoptosis in the ovary tissue (P<0.05), and transplanted cel s were able to survive in the ovary of nude mice. Al these findings show human umbilical cord blood mononuclear cel transplantation do have curative effects on premature ovarian failure in nude mice through the inhibition of apoptosis and the regulation of hormone secretion.

Objective To explore the effect of Chinese herbal medicine formula "tonifying kidney plus strengthening vital energy" on regulating CD40, CD40L protein and mRNA expression in spleen cell in mice with SHENXU. Methods The interaction of CD40L expressed by activated T cells with CD40 on monocytes, macrophages, or B cells is essential for the production of IL-12 and other cytokines. An experimental model of mice with SHENXU by corticosterone was established to observe the effect of Chinese formula on CD40, CD40L protein and mRNA expression in spleen cells or in ConA stimulated spleen cells by immunohistochemistry and RT-PCR. Results No significant differences were found among 5 groups of mice about the CD40 and CD40L protein expression in spleen cells. Both CD40 and CD40L expression detected by immunohistochemistry decreased seriously in ConA induced cells from SHENXU mice (18.9%?3.3% vs 15.8%?2.3%) as compared to that of normal controls(29.7%?4.2% vs 23.3%?1.3%), and so did the mRNA expression on ConA stimulated cells from SHENXU mice. They were restored in different degrees when the SHENXU mice were cured by different Chinese medicine. The effect of "tonifying kidney plus strengthening vital energy" was stronger than that of "tonifying kidney" and "strengthening vital energy" alone. Conclusions "Tonifying kidney plus strengthening vital energy" enhanced the expression of CD40, CD40L mRNA and protein, and this may be one of the mechanisms of improving the status of immune inhibition resulted by Corticosterone.

Objective To study the effect of Chinese herbs on human scalp hair growth in vitro. Methods Organ culture of human scalp hair follicles was used to study the effects of extracts of Crataegus cuneata, Ligustrum lucidum, Polyporus umbelatus and Bletilla striata mixture and ginseng saponin on hair growth. Results Low doses of mixture extracts(1.28 ?g/mL and 6.4 ?g/mL) markedly enhanced the hair growth and lengthened the period of hair growth, while high doses of mixture extracts(4 mg/mL and 20 mg/mL) sharply inhibited hair growth and shortened the period of hair growth. High doses of ginseng saponin (40 ?g/mL and 200 ?g/mL) also had an inhibiting effect on hair growth. Conclusion This observation suggests that extracts of Crataegus cuneata, Ligustrum lucidum, Polyporus umbelatus and Bletilla striata mixture can promote hair growth in vitro.

OBJECTIVETo observe the effects of electroacupuncture (EA) at different acupoints on apoptosis-related serum and expression of microRNA (miRNA) in rats with myocardial ischemia, so as to explore its mechanism of action.

METHODSA total of 48 male Wistar rats were randomly divided into a normal group, a model group, a Neiguan group and a acupoint compatibility group, 12 rats in each group. Isoprenaline hydrochloride (ISO) with a daily dose of 2 mg/kg was subcutaneously injected for 14 days to establish the myocardial ischemia model in the model group, Neiguan group and acupoint compatibility group. Rats in the normal group were subcutaneously injected with an equal volume of normal saline. After modeling, rats in the Neiguan group were treated with EA at "Neiguan" (PC 6), while rats in the acupoint compatibility group were treated with EA at "Guanyuan" (CV 4), "Zusanli" (ST 36) and "Neiguan" (PC 6). Rats in the normal group and model group were treated with immobilization, once day for 21 days. The contents of creatine kinase-MB (CK-MB), vascular cell adhesion molecule-1 (VCAM-1) and endothelin-1 (ET-1) in serum were detected by enzyme-linked immunosorbent assay (ELISA); apoptosis index (AI) of myocardial cells was detected by TUNEL method; the expressions of miRNA-1, miRNA-133, miRNA-208 and miRNA-499 were detected by real-time PCR method.

RESULTSCompared with the normal group, the serum CK-MB, VCAM-1 and ET-1 were significantly increased in the model group, Neiguan group and acupoint compatibility group (all P < 0.01), and the apoptosis index was significantly increased (all P < 0.01). The CK-MB, VCAM-1 and ET-1 in the Neiguan group and acupoint compatibility group were significantly lower than those in the model group (all P < 0.01); the AI was reduced, which was more significant in the acupoint compatibility group (P < 0.05). Compared with the normal group, the expression of miRNA-133 was reduced (P < 0.01) and those of miRNA-208, miRNA-1 and miRNA-499 were significantly increased in the model group (all P < 0.01). Compared with the model group, the expression of miRNA-133 was increased (both P < 0.01) and that of miRNA-208, miRNA-1 and miRNA-499 were significantly reduced (all P < 0.01) in the Neiguan group and acupoint compatibility group. Compared with the Neiguan group, the expression of miRNA-133 was increased (P < 0.01) and those of miRNA-208, miRNA-1 and miRNA-499 were significantly reduced in the acupoint compatibility group (P < 0.01, P < 0.05).

CONCLUSIONEA at acupoints, especially acupoint compatibility group, could effectively prevent and treat myocardial ischemia, and the protective effect is possibly correlated to the double regulation on increasing the expression of miRNA-133 and inhibiting the expression of miRNA-1, miRNA-208, miRNA-499.

Acupuncture Points ; Animals ; Apoptosis ; Disease Models, Animal ; Electroacupuncture ; Endothelin-1 ; genetics ; metabolism ; Humans ; Male ; MicroRNAs ; genetics ; metabolism ; Myocardial Ischemia ; genetics ; metabolism ; physiopathology ; therapy ; Myocytes, Cardiac ; metabolism ; Rats ; Rats, Wistar

Aim To investigate the time-dependent effect of insulin on the expression of SREBP-1(sterol regulatory element binding protein-1),FAS(fat acid synthase)and lipid droplet formation in HKC cells(human renal proximal tubular epithelial cells line).MethodsHKC cells were respectively treated with 100 nmol·L~(-1) insulin for 0,2,4,6,12 h and 24 h.The analysis of SREBP-1 and FAS mRNA was performed by RT-PCR and the expression of SREBP-1 protein was detected by Western blot and immunocytochemistry.Furthermore,Oil Red O staining was used to determine cellular lipid droplet formation.ResultsCompared with HKC cells of 0 h group,there was no difference of SREBP-1 and FAS mRNA in HKC cells of 2 h group.However,the expression of SREBP-1 and FAS mRNA was significantly increased in HKC cells of 4,6 h and 12 h group.Further,the most expression of SREBP-1 and FAS mRNA was at 6 h group and was respectively increased by 3.578 and 4.272 times compared with 0 h group.The results of Western blot showed that the precursor and mature segments of SREBP-1 protein in 4,6 h and 12 h group HKC cells were increased and those of 6 h group HKC cells were the highest and about 4.106 and 5.167 times than those of 0 h group HKC cells.Immunocytochemistry presented the result that SREBP-1 protein was located in the plasma and the expression of 4,6 h and 12 h group HKC cells was significantly higher than that of 0,2 h and 24 h group HKC cells.The result of Oil Red O staining showed that lipid droplet markedly deposited in 6 h group HKC cells,contrarily,no lipid droplet was found in HKC cells of other groups.ConclusionAbove results suggested that insulin up-regulated SREBP-1 and FAS in time-dependent manner that led to cellular lipid droplet deposit,which may play an important role in the pathogenesis of renal lipid accumulation in metabolism syndrome.

OBJECTIVETo compare the clinical effects between ginger-partition moxibustion combined with glucocorticoid and simple oral glucocorticoid for thyreoitis at subacute stage.

METHODSEighty-one patients were randomly divided into an observation group (41 cases) and a control group (40 cases). In the observation group, ginger-partition moxibustion and hormone were applied. Moxa cones were used at local ashi points, Zusanli (ST 36), Guanyuan (CV 4) and Qihai (CV 6), six cones every point, once every other day and three times a week. Besides, 24 mg methylprednisolone tablets were adopted orally every day, and in two weeks the dose was 16 mg/d, in four weeks 8 mg/d, in six weeks 4 mg/d; all the patients were observed for 8 week. In the control group, simple methylprednisolone was prescribed orally, and the dose, the usage and treatment time were the same as those in the observation group. Erythrocyte sedimentation rate (ESR), triiodothyronine (T3), thyroxin (T4) and ultra-sensitive thyroid stimulating hormone (TSH) before and after treatment were observed in the two groups, as well as fever, the pain of thyroid gland, the regression time of swelling and adverse reaction.

RESULTSThe time of thyroid gland pain relieved of the observation group was earlier than that of the control group [(3.07 ± 0.78) days vs (3.62 ± 0.92) days, P < 0.05]. After treatment, T3, T4 and ESR were declined apparently (all P < 0.01), and TSH was obviously increased in the two groups (both P < 0.01). After 2-week treatment, ESR in the observation group was lower than that in the control group (P < 0.05). After 4-week treatment, T3, T4, TSH and ESR in the observation group were better than those in the control group (all P < 0.05). After 8 weeks, all indices in the observation group were superior to those in the control group, without statistical significance between the two groups (all P > 0.05). The effects of the observation group in 2 weeks, 4 weeks and 8 weeks were better than those in the control group (all P < 0.05). The cured course was shorter in the observation group than that in the control group (P < 0.05). The adverse reaction rate in the observation group was lower than that in the control group [4.9% (2/41) vs 22.5% (9/40), P < 0.05]. Three months later after treatment, the cured patients were followed. There was no recrudescence in the observation group and three patients caught the disease again in the control group, but there was no statistical significance between the two groups (P > 0.05).

CONCLUSIONGinger-partition moxibustion combined with glucocorticoid achieves better effect than simple oral glucocorticoid for thyreoitis at subacute stage, and the adverse reaction is less, which presents the clinical advantages of the integration of Chinese and western medicine.

Adult ; Female ; Ginger ; chemistry ; Glucocorticoids ; administration & dosage ; Humans ; Male ; Middle Aged ; Moxibustion ; Thyroid Gland ; drug effects ; metabolism ; Thyroid Hormones ; metabolism ; Thyroiditis ; drug therapy ; pathology ; therapy ; Thyrotropin ; metabolism ; Young Adult

Objective To investigate the role of nuclear factor-kappa B (NF-κB)signaling pathway in propofol-induced suppression of up-regulation of inducible nitric oxide synthase (iNOS) gene expression in LPSstimulated RAW264.7 cells.Methods RAW264.7 cells were purchased from cell bank of Chinese Academy of Sciences and cultured in DMEM culture medium containing 10％ fetal bovine serum.The cells were seeded in 6 cm diameter dishes (3 ml/dish) or in 6-well plates (2 ml/well) with a density of 5 × 105/ml and randomly divided into 3 groups ( n =18): normal control group (group C),group LPS (group L)and group LPS + propofol (group LP).The cells were incubated with LPS 1 μg/ml in groups L and LP.Propofol 50μmol/L was added to the culture medium at 2 h before LPS in group LP.Cells were harvested at 30 min after being stimulated with LPS.Phosphorylation of IκB kinase(p-IKK) and NF-κB activity were detected by Western blot.The expression of iNOS mRNA was determined after 6 h exposure of the cells to LPS.Results LPS significantly up-regulated the expression of p-IKK and iNOS mRNA and increased NF-κB activity in group L as compared with group C.Propofol pretreatment significantly attenuated the effects of LPS on p-IKK,iNOS mRNA expression and NF-κB activity.Conclusion NF-κB signaling pathway is involved in the propofol-induced suppression of up-regulation of iNOS mRNA expression in LPS-stimulated RAW264.7 cells.

Objective To observe the synthesis of melanin and tyrosinase activity of the cultured normal human melanocytes after treated with baicalin and NB-UVB, and to provide theoretical evidence for study on the new therapy method of vitiligo.Methods The normal human menlanocytes cultured in vitro were divided into control group,different concentrations of baicalin groups,different doses of NB-UVB group and baicalin combined NB-UVB group.The proliferation rate of melanocytes was measured by MTT assay,the level of melanin was tested by NaOH pyrolysis,and the tyrosinase activity was measured by dopa oxidization.Results Compared with control group,the proliferation rates in 10-4 mol·L-1 baicalin group and 30 mJ·cm-2 NB-UVB group had no significant difference(P>0.05 ). Compared with control group, the levels of melanin and tyrosinase activities in baicalin group,NB-UVB group and combined group were increased (P<0.05 or P<0.01);compared with baicalin group and NB-UVB group,the level of melanin and tyrosinase activity in combined group were increased(P<0.01). Conclusion Baicalin combined with NB-UVB has better promotion effect on melanin synthesis than used alone.

Objective:To treat the mixed and deeper infantile hemangioma with ladder dosage propranolol, and to explore its efficacy and safety.Methods:A total of 98 infants with hemangioma were treated by ladder treatment of propranolol.Before treatment,comprehensive assessments of electrocardiogram(ECG),heart color ultrasound, blood glucose,liver function,kidney function,myocardial enzymes and blood routine were conducted.After excluding contraindications,the dose of propranolol incrementally doubled from 0.5 mg·kg-1·d-1 to 4.0 mg·kg-1·d-1.Propranolol was taken 3 times a day.Before and after medication for 1 and 2 h,ECG was monitored.The changes of tumor size,texture,color and other changes or an onset of adverse reactions were dynamicly observed.The infants were visited every month.The efficacy was evaluated using Achauer system.Results: After medication,98 cases had different degrees of color changes or tumor consistency softening.After the dosage of propranolol was increased to 4.0 mg·kg-1·d-1,the change of tumor was the fastest.According to the 4-grade method, there were 84 cases(85.71%) as gradeⅣ (excellent),2 cases (2.04%) as grade Ⅲ (good),4 cases (4.08%) as gradeⅡ (medium)and 8 cases (8.16%) as gradeⅠ (poor).The curative effect of mixed hemangioma was better than that of deeper hemangioma(P<0.05).The recovery time of 74 cases of hemangiomas was 6 months.The major adverse reactions were heart rate decline(5/98,5.10%),drowsiness(3/98,3.06%),diarrhea(7/98,7.14%),loss of appetite (1/98,1.02%), and convulsions (2/98,2.04%).After treatment,all adverse reactions disappeared.Two months after drug withdrawal there were 4 cases of recurrence,and they were continously treated with propranolol.Conclusion: The efficacy of oral ladder dosage propranolol in treatment of mixed and deeper infantile hemangioma is increased significantly and there are no significant adverse reactions.

Objective:To investigate the expression of mmp-2 in mesangial proliferative glomerulonephritis(MsPGN) rat renal tissue and the effects of nourishing kidney and activating blood flow recipe of TCM. Methods: 54 Male SD rats were divided into control group, MsPGN group and treatment group with nourishing kidney and activating blood flow recipe of TCM. Immunohistochemical staining, flow cytometry, western blot and RT-PCR were used to check the protein expression of mmp-2. Results: In MsPGN group, following with the disease progressing, the glomerulus grew graduatly, ECM increased, the basement membrane of glomerulus was thicker. The immunohistochemical staining showed that the chief positive granules were deposited in glomerulus and renal tubular, the expression of mmp-2 was lower than that in control group, but in treatment group the expression of mmp-2 was higher than that in MsPGN group. Conclusion: In MsPGN renal tissue, the expression of mmp-2 was reduced, its activity was weakened. Nourishing kidney and activating blood flow recipe of TCM could induce the expression of MMP-2 and partly recover the activity. Nourishing kidney and activating blood flow recipe of TCM could resist glomerular sclerosis and postpone the course of chronic renal failure.