ObjectiveTo assess the microscopic features of alopecia areata(AA) by using in vivo confocal laser scanning microscopy(CLSM).MethodsBetween January 2010 and May 2011,26 male and 20 female patients diagnosed with AA were enrolled in this study.AA lesions and perilesional normal skin were examined by means of in vivo CLSM.Tissue specimens were obtained from the lesions and perilesional normal skin of 10 out of the 46 patients and subjected to routine histopathologic examination.ResultsThe number of hair follicles per unit area(cm2) was decreased in lesions of progressive AA and resting AA compared with normal perilesional skin(134.856 ± 18.301 and 147.159 ± 17.536 vs.301.613 ± 35.317,both P ＜ 0.05).Although the quantity of hair follicles increased in lesions of recovery AA((227.778 ± 16.861 )/cm2),but was still less than that in the normal perilesional skin (P ＜ 0.05).There was a lack of hair shaft in follicles,as well as an inflammatory infiltration in hair follicles,around hair follicles and capillaries in superficial dermis in lesions of progressive AA.The inflammatory infiltration was attenuated in lesions of resting AA.In lesions of recovery AA,the infiltration was further attenuated with an apparent growth of lanugos and terminal hairs.ConclusionsAs far as AA lesions are concerned,CLSM images are consistent with histopathological findings.CLSM may serve as a promising tool for monitoring efficacy and predicting prognosis based on hair growth cycle,hair follicle number,and inflammatory infiltration degree.

Objective To investigate the correlation between human leukocyte antigens-A,-B,-DRB1 (HLA-A,-B,-DRB1) high resolution alleles and chronic renal failure (CRF) caused by immunoglobulin-a nephropathy (IgAN).Method The polymerase chain reaction-sequence-based typing (PCR-SBT) method was used to investigate the genotypes of HLA-A,-B and-DRB1 high-resolution alleles in 191 cases of CRF caused by IgAN (experimental group) and 503 healthy blood donors (control group).The alleles frequencies between two groups were compared and the association between CRF caused by IgAN and the polyrnorphism of HLA was analyzed.Result (1) There were 25 alleles at A locus,48 alleles at B locus and 32 alleles at DRB1 locus in experimental group.(2) The genetic frequency of HLAA * 2901 [Pc =0.033,OR =10.738,95％ CI (1.193,96.691)],HLA DRB1 * 1106 [Pc =0.0001,OR =0.969,95％ CI (0.944,0.994)],HLA-DRB1 * 1202[Pc =0.002,OR =1.859,95％ CI (1.259,2.745)],HLA-DRB1 * 1401 [Pc =0.021,OR =0.984,95％ CI (0.967,0.998)],HLA-DRB1 * 1602[Pc=0.015,OR=1.915,95％ CI (1.157,3.17)] in experimental group was higher than in control group (P＜0.05).Conclusion There is susceptibility association of HLA-A * 2901,HLA-DRB1 * 1106,HLA-DRE * 1202,HLA-DRB1 * 1401,HLA-DRB1 * 1602 with CRF caused by IgAN.It is concluded that there is a close genetic and immunological correlation between HLA alleles and the pathogenesis of CRF caused by IgAN.

Objective To study the effect of sandalwood essential oil on isolated ileum smooth muscle of guinea pigs and the small intestine movement function of mice.Methods We used the experiment method of isolating ileum smooth muscle of guinea pigs and intestine propulsion of carbon ink in mice.We constructed the models of healthy mice,neostigminetrested mice and adrenaline-loaded mice.Results Sandalwood essential oil had an inhibitory effect on the spontaneous movement of guinea pigs isolated ileum and an antagonistic action on intestinal spasm caused by acetylcholine,histamine and barium chloride(P

Objective To investigate the influence of mannitol and glycerin injection (containing 15％ mannitol and 15％ glycerin) on experimental intracranial hypertension.Methods Rabbits were randomly divided into eight groups (n =8 rabbits/group):normal control group,model control group,mannitol-glycerol injection groups (2.5 ml/kg group,5 ml/kg group,and 10 ml/kg group),compound mannitol injection fluid group,20％ mannitol group,and 10％ glycerol-sodium chloride injection group.The continuous intravenous infusion of nitroglycerin [0.04 mg/(kg · min),10 min] on rabbits was used to establish intracranial hypertension model except the normal control group of animals,and effect of mannitol glycerol injection on it with a single intravenous injection was observed.Serum renal function,electrolytes and other indicators were tested.Results Intravenous infusion of mannitol glycerol injection (2.5 ml/kg,5 ml/kg,and 10 ml/kg) could significantly reduce rabbit nitroglycerin-induced intracranial hypertension,and dosedependent,with increasing dose reducing intracranial pressure could enhance(P ＜ 0.05).Mannitol glycerol injection (5 ml/kg) produced the same intracranial pressure compared to 20％ mannitol and compound mannitol injection,and maintained a long-time role.Conclusions Mannitol glycerin injection can significantly reduce intracranial pressure.Its intensity is the same as 20％ mannitol and compound mannitol injection,and maintains a longer intracranial pressure without significant renal dysfunction and electrolyte distnrhances.

Objective To establish animal models of functional dyspepsia with spleen deficiency and to compare the efficacy of different methods.Methods Rat models were established by iodoacetamide(IA)-treatment or combined with swimming.Appearance,body weight,food intake of the rats were observed,and serum motilin,cholecystokinin,lactate,gastrin content and urinary D-xylose excretion rates were detected to confirm whether the model of functional dyspepsia with spleen deficiency was established.Results The IA-treated rats had less food intake and a slower body weight gain.The IA-treated combined with swimming rats presented spleen-hypofunction symptoms,such as emaciation,hair dry and loose stools,their urinary D-xylose excretion rate,serum motilin,gastrin content were decreased,and serum cholecystokinin and lactate contents were increased significantly (P<0.05 for all).Conclusions All the three methods used in this study can result in symptoms of functional dyspepsia with spleen deficiency.However,IA-treatment combined with swimming models appear more close to spleen deficiency-like presentation,and the best model is the IA-treated combined with platform standing.

Objective To establish a mouse model of circulating tumor cells (CTCs) by applying mouse hepatoma Hapa 1-6 cells.Methods 108 healthy male C57BL/6 mice were randomly divided into 3 groups according to their body weights.Hepa 1-6 cell suspension was intravenously injected to each mouse in the three groups at a concentration of 1×106,5×106 and 1×107/mL,0.2 mL per mouse,respectively.Blood samples were collected from the mice on the 1st,5th,9th,13th,17th and 21st days after tumor cell injection.The number,ratio and relative inhibition rate of CTCs were calculated in 20,000 nucleated cells.The mortality of mice was recorded.②80 male C57BL/6 mice were averaged into 2 groups according to their body weight: control and sorafenib tosylate groups.0.2 mL of Hepa 1-6 single cell suspension was injected to each mouse through the caudal vein at a concentration of 5×106/mL.The mice were gavaged with sorafenib tosylate (50 mg/kg) for 21 days and blood samples were collected at the 3rd,8th,15th,and 21st days for CTC assessment.Results For the 1×106/mL group,the CTC inhibition rate was 25.1%,18.1%,8.9%,4.4%,2.9% and 0.3% on the 1st,5th,9th,13th,17th and 21st days,respectively,and all the mice were alive.For the 5×106/mL group,the CTC inhibition rate was 40.4%,35.4%,15.4%,9.0%,6.6% and 4.1% on the 1st,5th,9th,13th,17th and 21st days,respectively,and all the mice were alive.For the 1×107/mL group,the CTC inhibition rate was 39.1% and 33.5% on the 1st and 5th days,respectively.Some mice died immediately after intravenous injection and all mice died within 7 days.②The relative clearance of CTCs was-7.5%,4.6%,55.3% and-94.5% on the 3rd,8th,15th and 21st days of sorafenib tosylate administration.Compared with the control group,there were significant differences among the three groups (P<0.05 or P<0.01).Conclusions A mouse model of circulating hepatoma cells has been established by intravenous injection of 0.2 mL of 5×106/mL mouse Hepa 1-6 cell suspension.This mouse model can be used for screening and evaluation of drugs for circulating tumor cell inhibition.

Objective To study the effects of stratum corneum on the percutaneous absorption of fluorescein sodium (NaFl) liposome in rat skin. Methods Stratum corneum was stripped off by Scotch adhesive tape, and the NaFl content in the stratum corneum and the skin with no stratum corneum were detected by spectrofluorimetry at regular intervals. The dynamic permeated amount of different preparations of NaFl in the skin at different intervals in Franz diffuse cell was determined and the distribution of NaFl at 4 h after stripping off the stratum corneum was observed by fluorescence microscope. Results The concentrations of NaFl in the stratum corneum and the skin with no stratum corneum after applying liposome NaFl preparation were significantly higher than those of NaFl solution and gel preparations (P 0.05). Conclusions It is suggested that liposome can increase and change the penetration of NaFl into the stratum corneum and the skin with no stratum corneum in vitro. Hair follicular structure may not play an important role in the drug diffusion when stratum corneum is removed.

Objective To observe the improving effect of Hypericum Perforatum L Extracts (HPLES)on depression induced by chronic unpredictable mild stress in mice. Methods The depression model was established by the method of chronic unpredictable mild stress. Fifty depression model mice were divided into model control group,fluoxetine hydrochloride group (2. 6 mg / kg),Hypericum perforatum ex-tract low,medium and high (0. 2 g / kg,0. 4 g / kg,0. 8 g / kg) dose groups according to the random num-ber table method. Another 10 normal mice matched with body weight were taken as the normal control group. The mice in normal control group and the model control group were given pure water by gavage every day,and the mice in other groups were given corresponding solution by gavage for 4 weeks. In addition to the normal control group,the mice in other groups continued to undergo chronic unpredictable mild stress during gavage. The sugar water preference test and forced swimming test were performed after the last administration. Blood samples were collected from the posterior orbital venous plexus,and the levels of dopamine (DA) and brain-derived neurotrophic factor (BDNF) were measured by Elisa. The hippocampal tissues of mice were exam-ined by HE staining. Results Compared with the normal control group,the body mass of mice in the model control group decreased significantly at the first,second,third and fourth weeks ( t=2. 739,4. 162,4. 082, 3. 957;all P<0. 05). At the first,second,third and fourth weeks,the body mass of mice in the low,middle and high dose group of Hypericum perforatum extract were not significantly different from those in the model control group (all P>0. 05). Compared with the normal control group,the sugar water preference index of mice in the model control group was significantly reduced((61. 3± 4. 5)%,(52. 6± 5. 2)%; t=2. 721,P<0. 05),the swimming immobility time was prolonged(( 44. 3± 20. 00) s,(101. 8± 50. 8) s;t=2. 939,P<0. 05),the difference were statistically significant. Compared with the model control group,the sugar water preference index of mice in the low,middle and high dose group of Hypericum perforatum extract increased ((61. 8±4. 7)%,(65. 2±4. 1)%,(62. 6±5. 6)%,t=-3. 005,5. 073,-2. 928,all P<0. 05),the swimming immobility time decreased ((47. 2±17. 9) s,(54. 8±50. 3) s,(61. 3±44. 2) s; t=2. 803,1. 921,1. 903,all P<0. 05). The results of Elisa showed that compared with the normal control group,the levels of serum DA and BDNF of mice in the model control group were significantly lower (t=3. 031,8. 507,all P<0. 05); com-pared with the model control group,the levels of serum DA of mice in the low dose and high dose group of Hypericum perforatum were significantly higher (t=5. 025,3. 414,P<0. 05),and the serum BDNF of mice in the high dose group of Hypericum perforatum was also significantly higher (t=6. 098,P<0. 05),the differ-ence was statistically significant. HE staining showed that compared with the normal control group,the neu-rons in CA3 area of hippocampus in the model control group mice were seriously damaged,suggesting the es-tablishment of the mouse model. Compared with the model control group,the atrophy and degeneration of hippocampal CA3 cells in the three dose groups were significantly reduced. The atrophy and deformation of hippocampal CA3 neurons in the low,middle and high dose groups of Hypericum perforatum extract were re-lieved. Conclusion HPLES have obvious improving and antidepressant effects on the depression model mice induced by chronic unpredictable stress. The above effects may be related to the improvement of serum DA,DBNF level and reduce neuronal damage in CA3 area.

Postpartum depression(PPD)is one of the most common types of postpartum psychiatric syndromes. Because of the complex and changeable characteristics in PPD disease and the special period after childbirth, there are many clinical limitations in the study of this disease. Therefore,the preparation and establishment of a proper animal model closed to clinical and behavioral evaluation method plays an important role in study of its pathogenesis. This review mainly introduces the commonly used postpartum depression animal models and the behavioral evaluation method. It is hoped to provide a reference for further study of PPD pathogenesis and for the drug research and development.

Objective To study the protective effect of shenfu injection on cerebral ischemia reperfusion injury in rats.Methods 120 male Sprague Dawley (SD) rats(320-350 g) were randomly divided into sham operation group,model control group,Nimodipine injection group,low,medium and high dose group of shenfu injection according to gender weight.20 males in each group were given medicine once a day for 7 days before operation.The cerebral ischemia model was established by thread embolization after 5 days of administration.In the sham operation group,the other operations were the same as those in the model group except for carotid artery ligation and thread insertion.After 24 hours of perfusion,the neurological score,abdominal aorta blood flow,malondialdehyde (MDA),superoxide dismutase (SOD) and glutathione (GHS) levels in brain tissues were measured.Triphenyltetrazolium chloride (TTC) staining was used to calculate the area of cerebral infarction and pathological examination of brain tissues.Results Compared with the model control group,the middle and high dosage of shenfu injection could obviously improve the nerve function and increase the percentage of cerebral infarction area (P ＜ 0.05);the high dosage group of shenfu injection could obviously decrease the whole blood viscosity (P ＜ 0.01);the middle and high dosage of shenfu injection could obviously reduce the level of MDA in rat brain tissue (P ＜ 0.01) while increasing the levels of SOD and GSH (P ＜0.01),finally could significantly improve the pathological changes of brain tissues such as mild swelling of nerve fibers,mild neuronal degeneration,inflammatory interstitial edema and inflammation.Conclusions Shenfu injection has obvious protective effect on cerebral ischemia reperfusion model in rats.