OBJECTIVE:To study effect of liposome suspension on distribution of fluorescein sodium in the rat skin.METHODS:Using0.125%fluorescein sodium(NaFl)solution as control,the distribution of NaFl in epidermis and dermis at different time periods was observed under confocal laser scanning microscope in0.125%NaFl liposome suspension group(trial group).RESULTS:The fluorescent intensity peaks in epidermis and dermis of trial group were higher than those in skin layers of control group(P

Obese gene was cloned and its protein product which named leptin was found to be expressed specially in fat tissues in 1994. As a metabolic signal of reproductive system, leptin reflects the situations of nutrition and energy, which the body supplys to the brain, and stimulates reproductive endocrinology system, then regulates the functions of reproduction through hypothalamic-pituitary-gonadal axis.The present status of eptin in this field was reviewed.

Obese gene was cloned and its protein product which named leptin was found to be expressed specially in fat tissues in 1994. As a metabolic signal of reproductive system, leptin reflects the situations of nutrition and energy, which the body supplys to the brain, and stimulates reproductive endocrinology system, then regulates the functions of reproduction through hypothalamic-pituitary-gonadal axis. The present status of leptin in this field was reviewed.

OBJECTIVE: To observe the release mode and the amount of retention in skin of podophyllotoxin liposomes in topical application. METHODS: Two kinds of podophyllotoxin liposome suspension (DPPC liposome and bean lecithin liposome) and podophyllotoxin tincture in same concentration were topically applied on the skin of porkets and the amounts of retention of drug in skin of different preparations were detected with confocal laser scanning microscope.RESULTS: The quantity of retention of podophyllotoxin DPPC liposome in epidermis and superficial layer of dermis was larger than those of other two preparations and the drug concentration in applying DPPC liposome kept higher for 48 hours.CONCLUSION:Podophyllotoxin liposome coated by DPPC is much more targetable to skin and is a fairly good topical preparation for applying on skin.

Objective: To apply D101 macroporous resin for separation and purification of triptolide. Methods: After extracted by alcohol the extracts were dissolved in water and passed through a macroporous resin column and a aluminum oxide column. Then the triptolide on the columns were eluated with 95% alcohol and determination by HPLC. Results:Triptolide could be retained on D101 column. Conclusion: D101 resin can be used to refine the triptolide.

Objective To investigate changes in the expression of prepro-orexin and orexin receptor-1 ( OX1R) following permanent middle cerebral artery occlusion ( MCAO ) with or without preconditioning through electrical stimulation of the cerebellar fastigial nucleus (FNS). Methods Wistar rats were subjected to permanent MCAO and randomly divided into 5 groups: a sham-operated control group (PO), an FNS preconditioning + shamoperated control group (FNS-PO) , an ischemia group, an FNS preconditioning + ischemia group (FNS-PI) and a cerebellar fastigial nucleus injury + FNS preconditioning + ischemia group (FNL-FNS-PI). Each group was divided into 5 subgroups according to the time at which the animals were sacrificed after the MCAO ( 1, 3, 6, 12 and 24 h).RT-PCR was used to detect expression of OX1R mRNA, and ELISA to measure the levels of orexin-A in the hypothalamus and plasma. Results The immunoreactivity of prepro-orexin decreased significantly in the PI groups, with further decreases over time. At the 12th h after MCAO, the immunoreactivity of prepro-orexin reached a minimum.There were significant differences between the rats in the PO and FNS-PO groups. On the contrary, the immunoreactivity of OX1R increased significantly in the PI groups, with further increases continuing over time, peaking at 12 h after the MCAO. There were significant differences between the PO and FNS-PO groups. In the rats with FNS preconditioning (PI-FNS) , the decrease in prepro-orexin and the increase in OX1R were significantly inhibited compared to the PI subgroups at the 6th and 12th hour. There was no significant difference between the FNL-PIFNS group and the PI group. The expression of OX1R mRNA increased significantly in the PI group, with further increases continuing over time, peaking at 24 hours. The plasma levels of orexin-A were not significantly different among the groups, but the levels of orexin-A in the hypothalamus decreased significantly in the PI and FNL-PI-FNS groups, with further decreases continuing over time. At the 12th h after the MCAO the levels were significantly different compared with the PO and PO-FNS groups. While in the rats with FNS preconditioning (PI-FNS) , the decrease in orexin-A level was reversed and there was no significant difference compared with PO and PO-FNS groups. Conclusions The orexinergic system is altered following cerebral ischaemia. FNS preconditioning may be able to regulate these changes.

Objective To test the efficacy of low-frequency electrical acupoint stimulation plus rehabilitation training in improving swallowing function after stroke. MethodsEighty dysphagic stroke patients were randomly allocated into 2 groups of 40.Patients in the experimental group received low-frequency electrical acupoint stimulation plus 2 courses of rehabilitation training.Control group patients received the same protocol,but the electrical stimulation was not over acupoints.Their swallowing function was assessed using the Kubota water drinking test.ResultsAfter treatment,both groups showed significantly lower Kubota scores compared with pre-treatment.Prepost test difference was 1.25 ± 0.78 in the experimental group and 0.70 ± 0.72 in the control group,a significant difference.The total recovery rate in the experimental group was 87.5％.In the control group it was 55％.The improvement in swallowing function was significantly better in the experimental group.Conclusion Both groups showed a significant improvement in swallowing function,but low-frequency electrical acupoint stimulation increased the effectiveness of the rehabilitation training.

AIM: To investigate the role of autophagy in the apoptosis of human pulmonary artery endothelial cells (HPAECs) induced by cigarette smoke extract (CSE).METHODS: HPAECs were cultured routinely.HPAECs were treated with CSE at different concentrations, and the cell viability was detected by MTT assay.HPAECs were divided into control group, CSE group, 3-methyladenine (3-MA) group and 3-MA+CSE group.The autophagy was observed under fluorescence microscope with monodansylcadaverine (MDC) staining.Annexin V/propidium iodide staining and Hoechst 33342 staining were employed to detect apoptosis.In addition, the protein levels of LC3, beclin-1 and cleaved caspase-3 were determined by Western blot.RESULTS: MDC staining showed the increased production of autophagic vacuoles was observed in CSE group.The results of Western blot showed that the expression levels of autophagy-related proteins LC3 and beclin-1 were increased, while 3-MA pretreatment inhibited the expression of these proteins and the production of autophagic vacuoles.Observation with Annexin V/propidium iodide staining and Hoechst 33342 staining showed that the apoptotic rate in CSE group was significantly higher than that in control group, and pretreatment with 3-MA induced further increase in the cell apoptosis.The protein level of cleaved caspase-3 in CSE group was significantly higher than that in control group (P<0.05), and 3-MA+CSE treatment induced the further increase in the protein level of cleaved caspase-3.CONCLUSION: CSE induces autophagy and apoptosis in the HPAECs.Inhibition of autophagy promotes the apoptosis induced by CSE in HPAECs, which can be achieved through activation of caspase-3.

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 find a rapid and simple method for quantifying the amount of eluted proteins in chromatography. Methods The method was developed by utilizing the principle of the absorbance of protein at 280 nm and the integral function of the software UNICORN for AKTA FPLC, namely, the protein quantity was evaluated based on its A_ 280 and peak area. Results The relation between peak area and quantity of the protein was linear correlation. The protein quantity could be expressed as X=Y/A. The Y represented the peak area (mAU?ml) of the interested fraction, and A represented the measurable value (mAU) of 1 g/L interested protein through the FPLC 280 nm cell and X was the quantity of the interested protein (mg). The method was used in the purification of recombinant human zona pellucida-3 protein (rhZP3)and the quantity of the purified rhZP3 evaluated from the formula was confirmed by Lowry method. Conclusion The method is easy, rapid, repeatable and practicable to quantify the protein in chromatography.