[Objective]To probe the approach of supplementing Qi and nourishing Yin in the treatment of lung cancer. [Methods] With detailed research into Chinese medicine literature from recent years regarding its application in lung cancer, the basic pathogenesis of lung cancer, and relevant clinical effects and fundamental research of lung cancer are summarized.[Result]Numerous studies have shown that Qi and Yin deficiency is the basic pathogenesis of lung cancer, thus the approach of supplementing Qi and nourishing Yin can reduce the side effects and strengthen the clinical effects of radio-chemotherapy, stim-ulate immune function, improve life quality, and prolong the survival time of the patients.[Conclusion]It is significantly obvious that the approach of supple-menting Qi and nourishing Yin has shown great advantages in the treatment of lung cancer, however further clinical and basic research are expected.

Lipoxins are metabolic products of arachidonic acid,which possess a wide spectrum of adjustment function for various inflammatory cell function and the expression of inflammatory related genes.It is known asstop signals or braking signals of inflammatory response,which can promote the regression of inflammation through regulating a variety of inflammatory signaling pathway.Now,the progress in the regulation of multiple signal pathways by lipoxins and its anti-inflammatory mechanism are reviewed.

Objective To investigate the differentiation from human mesenchymal stem cells (hMSC) into neuron-like cells. Methods hMSC were separated from rib marrow with Ficoll-Paque reagent and expanded in culture medium. hMSC were induced to differentiate into neurons with DMEM/BHA/DMSO or DMEM/monothioglycerol, respectively. Neuron-specific enolase (NSE), neurofilament (NF), nestin, glial fibrillary acidic protein (GFAP) were detected by immunohistochemistry. Results hMSC were expanded to be undifferentiated cells in culture for more than 10 passages. The isolated and cultured MSC comprised a single phenotypic population and displayed a fibroblast-like morphology. Simple method induced hMSC exhibiting a neuronal phenotype, with a positive expression of NSE, NF-M and nestin at 5 hours. But the neuron-like cells did not express the glial astrocyte marker GFAP. Conclusion It suggests that hMSC can be differentiated into neurons in vitro .

Objective To established a method for the content determination of ginsenoside Rg1 in Yushangling Capsules by HPLC. Methods HPLC was used with the C18 column. The mobile phase was acetonitrile-0.5% phosphoric acid (24∶76). The flow rate was 1.0 mL/min, the column temperature was 25 ℃, and the detection wave was set at 205 nm. Results The calibration curve was linear in the range of 0.05～0.80 ?g. The regression equation was Y =3629375.5X+2517.1, r =0.9998. The average recovery was 100.48% and RSD was 1.59%. Conclusion The method is simple, accurate and suitable for the content determination of ginsenoside Rg1 in Yushangling Capsules.

BACKGROUND: Nuclear factor kappa B (NF-κB) is possibly related to regulation of various cell signals that are derived from aqueous uveoscleral outflow pathway.OBJECTIVE: To explore effect of travoprost on the expression of NF-κB and inhibitor-κB (I-κB) in human ciliary muscle cells cultured in vitro. DESIGN, TIME AND SETTING: A contrast study, which was performed in the Laboratory of Zhongshan Ophthalmology Center from March 2005 to November 2006.MATERIALS: Eyeballs were obtained from the youth who died due to other diseases except eye disease no more than one hour. The relatives voluntarily provided the informed consent.METHODS: Travoprost (1 μmol/L) was added in human ciliary muscle cell culture medium, and then the samples were divided into four groups according to culture time, including 0-hour (control group), 6-hour, 12-hour, and 24-hour experimental groups. MAIN OUTCOME MEASURES: Expression of mRNA and protein of NF-κB p65 and I-κBα in the four groups by using real-time RT-PCR, immunofluorescence relative quantitative analysis and enzyme linked immunosorbent assay (ELISA) techniques. RESULTS: As compared to control group, mRNA expression of NF-κB p65 in the 6-hour, 12-hour, and 24-hour experimental groups was decreased (F=17.068, P=0.001); while mRNA expression of I-κBα was not changed remarkably in the 6-hour and 12-hour experimental groups (P > 0.05), but the expression was significantly higher than that in the 24-hour experimental group (F=32.742, P=0.000). Immunofluorescence relative quantitative analysis showed that the fluorescence intensity of NF-κB p65 in the 6-hour, 12-hour, and 24-hour experimental groups were weaker than that in the 0-hour control group (F=17.216, P=0.000); additionally, as compared to 0-hour control group, fluorescence intensity of I-κBα in the 6-hour experimental group was not changed remarkably (P=0.134), that in the 12-hour experimental group was weakened (P=0.032), and that in the 24-hour experimental group was strengthened (F=17.346, P=0.001). ELISA revealed that expression of phosphorylated NF-κB p65 was decreased gradually by the time of being induced by travoprost (F=15.4, P=0.001). CONCLUSION: Travoprost can down-regulate mRNA expression of NF-κB p65, inhibit nuclear translocation, and up-regulate mRNA expression of I-κBα in human ciliary muscle cells.

BACKGROUND:At present, there is still lack of related reports about the aging process of in vitro cultured epidermal cells, since epidermal cells are seed cells necessary for the construction of tissue engineered skin, this articleis is aimed to investigate the biological property of normal human epidermal cells during aging process so as to provide a foundation for the selection of seed cells for tissue engineered skin OBJECTIVE: To observe the in vitro proliferation and aging property of human epidermal cells in order to provide a foundation for the proper selection of seed cells for tissue engineered skin.DESIGN: A self-comparative experiment.SETTING: Orthopedic Surgery Research Instioute of Weifang Medical College and the General Surgery Department of Weifang Medical College Affiliated Hospital.MATERIALS: This experiment was carried out at the Orthopedic Surgery Research Institute of Weifang Medical College, between September 2000and September 2002. Healthy foreskin tissue was obtained from 20 normal boys of 6-8 years old who received peritomy at the General Surgery Department of Weifang Medical College Affiliated Hospital.METHODS: Epidermal cells were obtained from normal young people for subculture. Cells were collected from different culture passages and taken as subjects, and their aging characteristics were assessed through morphological observation, population doubling time (PDT), immune cytochemistry and beta-galactosidase staining. MAIN OUTCOME MEASURES: ① The changes of the epidermal cell growth characteristics. ② The morphological changes of the epidermal cells. ③ The epidermal cell phenotypic changes. RESULTS: ① The clanges of the epidermal cell growth characteristics: Cells were in vitro cultured by monolayer for 9 passages, and PDT of P2 was the shortest. The cells showed strong proliferation in the first 5 passages.From P6, PDT was obviously prolonged, but the cells from P8 did not proliferate any longer. ② The morphological changes of epidermal cells: The primary cultured cells began to proliferate 3 days later, which accelerated 4 days later. The cells became approximately fused in about 1 week. The growth of epidermal cells was identified with a microscope and the immuno histological techniques. ③ The epidermal cell phenotypic changes: Along with the consecutive subculture, histological expression of beta-galactosidase was found to show an increasing tendency from weak expression (occupying 9% of the young cells) to strong expression (occupying 65% of aging cells), and the positive expression rate of beta-galactosidase was found to be remarkably correlated with cell passage age (r=0.87, P ＜ 0.01). CONCLUSION: ① Compared with young cells, aging cells displayed more obvious aging morphology and enzyme cyto-chemical characteristics.During the cell aging process, the PDT of cells showed an increasing tendency. ②Compared with young cells, the expression of beta-galactosidase in aging cells was remarkably increased, and this increase paralleled with the appearance of cell aging phenotype and the loss of cell proliferation capability, and reflects the aging degree of cells. ③ The in vitro cultured normal human epidermal cell aging model was established in this experiment. The results of this experiment indicated that epidermal cells from the 1st -5th passage (donators aged 16-18 years old) can be taken as the optimal seed cells for tissue engineered skin construction.

BACKGROUND: Fibroblasts are considered as seed cells necessary for the construction of tissue engineering skin. The ultrastructure of cells of various generations was observed under the electron microscope in the hope of providing foundation for proper selection of seed cells for tissue engineering skin.OBJECTIVE: To observe the ultrastructural changes of normal human fibroblasts during in vitro culture.DESIGN: Self-control observation.SETTING: Institute of Plastic Surgery, Weifang Medical College; Department of General Surgery affiliated to Weifang Medical College.MATERIALS: This experiment was carried out in the Institute of Plastic Surgery, Weifang Medical College, between September 2000 and September 2002. Healthy prepuce specimens were collected during posthetomy from normal boys aged 6-8 years after the informed consent was obtained from their guardians.METHODS: The normal human diploid fibroblasts were used to carry out consecutive subculture; cells were collected from different generations for morphological and ultrastructural observation under the inverted phase contrast microscope and transmission electron microscope.ture under the transmission electron microscope.microscope: Cells could pass on for 65-70 generations and survive for 280-300 days. Cells within 45 generations could grow rapidly, but gradually grew slowly after the 45th generation, and even displayed no proliferaunder the transmission electron microscope: There were no obvious changes in cell ultrastructure within 40 generations, but cells presented inward tolds of nucleus membrane from the onset of generations 41-65, with the ratio of cell nuclear/plasma reduced as well as cell surface process and microvilli also reduced.CONCLUSION: The ultrastructural change of in vitro cultured fibroblasts varied between different generations, which became obvious after the 41st generation, suggesting that fibroblasts within 40 generations are considered preferable seed cells for the construction of tissue engineering skin.

BACKGROUND: As the seed cells for construction of tissue engineered skin, fibroblasts directly decide the quality of tissue-engineered skin. During in vitro culture, collagen gene expression and response to epidermal growth factor (EGF) stimulation of the fibroblasts in different passages can be indicative of their proliferative capability for use as the seed cells for skin tissue engineering.OBJECTIVE: To observe the expression of type Ⅰ and Ⅲ collagen mRNA in fibroblasts cultured in vitro and fibroblast response to EGF stimulation, and thereby providing reference for the selection of optimal seed cells for tissue engineering.DESIGN: Self-controlled experiment.SETTING: Institute of Plastic Surgery, Weifang Medical College.MATERIALS: This experiment was carried out at the Institute of Plastic Surgery, Weifang Medical College between September 2000 and June 2002. The specimens of normal prepuce tissues excised by circumcision were obtained from 20 healthy boys at the age between 6 and 8 years on a voluntarily basis in the Department of General Surgery, Affiliated Hospital of Weifang Medical College.surgically excised prepuce by trypsin and type Ⅰ collagenase digestion. After cultured till 80% confluence, the cells were digested with mixed digescontrast microscope was used for dynamic observation of the cell morphology and growth status, and transmission electron microscopy and anti-vigen gene expression: Reverse transcriptase PCR (RT-PCR) was performed for amplification of type Ⅰ and Ⅲ collagen cDNA derived from the total sis of fibroblast response to EGF stimulation: The fibroblasts of P10 and P60passage were divided into treatment group with stimulation by the conditioned medium containing EGF and control group with treatment with only the conditioned medium. 3H-TdR incorporation assay was performed for analyzing the growth of the fibroblasts in response to EGF stimulation.lasts of different passages to EGF stimulation.decreased with cell passaging and 3H-TdR incorporation was lower in P60cells without significant difference between the treatment group and control group (132.5±23.6 vs 124.9±16.8, P ＞ 0.05) than in P10 cells with,however, significant difference between the two groups (512.8±56.4 vs 306.4±22.5, P ＜ 0.01).EGF stimulation is weaker than P10 cells, moreover additional EGF in the condition medium has no obvious regulation on the proliferation of P60cell growth, but extremely remarkable on P10 cells, implying along with the increase of cell passage, tritium-thymidine incorporation reduced and regulative capability of EGF on aging fibroblastic growth was also attenuated.

Objective To investigate the effect of RO20-1724 on the cognitive function in immature rats after ketamine anesthesia.Methods Ninety-six SD rats of both sexes,aged 21 days,weighing 45-55 kg,were randomly divided into 8 groups ( n =12 each):control group (group C) ; ketamine group (group K) ; ketamine + normal saline group (group K + N) ; ketamine + anhydrous alcohol group (group K + A) ; ketamine + 4 different doses of RO20-1724 groups (group K + R1-4 ).The rats were anesthetized with intraperitoneal injection of kctamine 70 mg/kg in groups K,K+N,K+A and K+.R1-4.Normal saline 2 ml,anhydrous alcohol (in normal saline 2 ml),and RO20-1724 0.25,0.50,0.75 and 1.00 mg/kg (in anhydrous alcohol 8 μl and then in normal saline 2 ml) were injected intraperitoneally in groups K + N,K + A and K + R1-4 respectively 30 min later.Six rats from each group were randomly selected at 24 h after administration and Morris water maze was used to test the ability of learning and memory.Six rats from each group were sacrificed at 48 h after administration and hippocampus and cerebral cortex were removed for determination of the expression of CREB and phospho-CREB (p-CREB) by Western blot.Ressults Compared with group C,the escape latency was significantly prolonged at 2-4 days after administration,the number of animals' swimming across the platform decreased,and the expression of CREB and pCREB in hippocampus and cerebral cortex down-regulated in groups K,K+ N,K+ E,K+ R1 and K+ R2(P ＜0.05 ).Compared with group K,the escape latency was significantly shortened at 2-4 days after administration,the number of animals' swimming across the platform increased,and the expression of CREB and p-CREB in hippocampus and cerebral cortex up-regulated in groups K + R3 and K + R4 ( P ＜ 0.05).Compared with groups K + R1 and K + R2,the escape latency was significantly shortened at 2-4 days after administration,the number of animals' swimming across the platform increased,and the expression of CREB and p-CREB in hippocampus and cerebral cortex up-regulated in groups K+ R3 and K+ P4(P ＜ 0.05).There were no significant differences in the escape latency,the number of animals' swimming across the platform,and the expression of CREB and p-CREB in hippocampus and cerebral cortex between groups K + R1 and K + R2,and between groups K + R3 and K + R4 ( P ＞ 0.05 ).Conclusion RO20-1724 0.75-1.00 mg/kg can improve ketamine-induced cognitive dysfunction by up-regulating CREB and p-CREB expression in hippocampus and cerebral cortex in immature rats.

Objective To investigate methylation of the P15INK4B and p27kipl genes in human mantle cell lymphoma cell line Mino,to evaluate the effects of decitabine on demethylation of p15INK4B and p27kip1 genes and on apoptosis of Mino cells and its relative mechanism. Methods Mino cells were after treated with various concentration of decitabine, the cell viability, cell cycle distribution or the apoptosis of Mino cells was respectively analyzed by trypan dye-exclusion assay or flow cytometry.The mRNA and protein expression of p15INK4B 、p27kip1 and bcl-2 were studied by RT-PCR or Western blot, respectively.Methylation of the p15INK4B and p27kip1 genes in Mino cells were determined by PCR using the methylation specific primer(MSP).Results Decitabine significantly inhibit the cell growth,induced G1 arrest and promoted apoptosis of Mino cells. The expression of p15INK4B and p27kipl mRNA were both significantly increased,wheres bcl-2 mRNA was decreased, After treatment with 6.4,3.2,1.6 mmol/L decitabine for 72 h,the methylationg of p15INK4B gene were decreased to 25.2 ％ 、48.2 ％ and 65.0 ％respectively,in the meantime,the methylationg of p27kip1 gene was decreased to 20.21％ 、50.2 ％ and 70.0 ％. Conclusion The hp15INK4B and p27kip1 genes of Mino cells are methylated and down-regulated.Decitabine can inhibit the proliferation and induce the apoptosis of Mino cell lines,with the reduction of bcl-2 gene and demethylation of p15INK4B and p27kip1 genes.