Objective To illustrate a method of lifting the upper-lip to alleviate the aging face,and to evaluate its effects and risks.Methods In the past two years,53 cases of overlong upper-lip were performed with this procedure.The recovery periods,lifted effects,side effects and the total satisfactions were evaluated.Results As for recovery periods,edema roughly reduced in 6-20 months,reddish scar faded in 1-11 months,expression restored in 2-8 months,and numbness disap peared from 4 to over 18 months.The lifted effects showed that 37 cases (78.7％) were satisfactory,7 cases (14.9％) of the lifting was not enough and still complained a long upper-lip,2 cases (4.2％) was ineffective,and 1 case (2.1 ％) considered an excessive lifting.The long term follow-up showed that 3 cases (6.4％) had obvious scar (visible at a distance of more than 1 m),14 cases (29.8％) had depression of columella/base of nostril alar or with increased exposure of nostril,3 cases (6.4 ％) had expansion or morphological changes of alar,6 cases (12.8％) had increased thickness of the upper lip and prominent vermilion tubercle,1 case (2.1％) had changes of expression,1 case (2.1％) had asymmetry,1 case (2.1％) had arching sagging,and 35 cases (74.5％) had numbness or insensitivity of upper-lip.Overall results showed very satisfied in 29 cases (61.7％),satisfied in 15 cases (31.9 ％) and dissatisfied in 3 cases (6.4 ％).Conclusions The technique for upper-lip lifting displays a significant effect with few complication.It is recommended for further clinical application.

Objective To establish HPLC fingerprint for evaluating and controlling the quality of Gastrodia elata. Methods HPLC Analysis and similarity calculation were used for establishing fingerprint chromatogram and classifying 18 different original samples; LC-MS and pure compound comparisons were employed for the fingerprint peaks identification. Results HPLC Fingerprint chromatogram was cons- tructed with 27 fingerprint peaks, 17 of which were common fingerpring peaks and 12 main compounds were identified. According to the similarity to standardize herbal medicine, 18 samples could be classified three groups, Anhui-Henan, Shanxi-Sichuan, and Yunnan habitats. The two critical points were 0.80 and 0.58 for correlation coefficient and 0.88 and 0.73 for cosine values. Conclusion The selected fingerprint peaks are diagnostic for G. elata and the constituted HPLC fingerprint chromatogram could be used for identifying different habitats and served as a powerful tool for further quality control of G. elata.

BACKGROUND:Bone marrow mesenchymal stem cells(BMSCs)have the properties involving high proliferation capability,widely distribution,functional tissue repair after injury,as well as immune modulation,by which bring us extensive therapeutic possibilities.There are plenty of methods for isolation of BMSCs,yet,BMSCs exhibit discrepancies in varied growth stage and culture conditions.Up to now,there has been no agreement about the identification methods for cultured BMSCs.OBJECTIVE:To review the isolation methods and biological characteristics of BMSCs,and to compare the differential expression of BMSCs between in serum and serum-free medium,prior to and after proliferation,as well as before and after induction.METHODS:A computer-based online search was performed using key words of "bone marrow mesenchymal stem cells,isolation,culture,induce,marker,and characterization" to find documents published in the database of CNKI (http://dlib.cnki.net/kns50/)or Pubmed(http://www.ncbi.nlm.nih.gov/PubMed)from January 2003 to June 2009.The languages were limited Chinese and English.A total of 237 literatures were searched by the computer.RESULTS AND CONCLUSION:The positive rates of CD44 and CD34 of BMSCs isolated by the whole bone marrow culture were smaller than that of the density gradient centrifugation.However,BMSCs isolated by the whole bone marrow culture were superior to those isolated by the density gradient centrifugation in cell viability,proliferation rate,confluence time,as well as generation time.Other methods for BMSCs isolation had drawbacks of large cost and high requirement of experimental equipments.Following conditions were used to identify BMSCs:cell adherence,cell surface molecule labeling,strong self-proliferation ability,as well as potentials multi-directional differentiation.BMSCs exhibit differential expression between in serum and serum-free medium,prior to and after proliferation,as well as before and after induction.

Z-Ligustilide, a major phthalide isolated from a widely used traditional Chinese medicine Ligusticum chuanxiong, possesses various pharmacological activities including neuroprotective, anti-inflammatory, antiproliferative and vasorelaxing effects. However, it is unstable and inclined to degrade in natural conditions, which limits its study and application greatly. In this study, degradation behavior of Z-ligustilide and its degradation products stored at room temperature under direct sunlight were investigated and structure elucidated by HPLC-UV, UPLC-QTOF-MS and NMR. Z-ligustilide degradation and total five degradation products were generated and detected. Two degradation products were unequivocally identified as senkyunolide I and senkyunolide H by comparison with reference compounds. Another two degradation products were further isolated by semi-preparative HPLC and structure elucidated as (E)-6, 7-trans-dihydroxyligustilide and (Z)-6, 7-epoxyligustilide by 1H and 13C NMR, respectively. The degradation pathways of Z-ligustilide were finally proposed. Oxidation, hydrolysis and isomerization are the major degradation reactions.

Objective: To detect methylation status of p16 CDKN2A exon 1 during experimental carcinogenesis in rats. Methods:Thirty male clean SD rats were fed with 0.02 g/L of 4-nitroquinoline-oxide (4NQO) in drinking water.13, 16 and 24 weeks after experiment the normal, moderate-severe dysplasia and invasive squamous cell carcinoma tissues were removed from their tongues respectively; then the methylation status of p16 CDKN2A exon 1 were detected by methylation-specific PCR(MSP). Results:A 123 bp-unmethylated product was amplified in all samples but the methylated product was not detected in any of the samples. Conclusion: The p16 CDKN2A exon 1 appeares to be unmethylated during carcinogenesis of tongue cancer in experimental rats.

Objective To investigate the biochemical properties of rat improved perfusion-acellular pancreatic bioscaffold (APB).Methods The fresh pancreas from 10 rats were perfused through portal vein.The histological structure,ECM composition and DNA quantification of APB were evaluated.For the biocompatibility study,a 0.5 cm2 APB construct was surgically placed within a dorsal subcutaneous pocket of mice.Results The pancreatic tissue become translucent and the macroscopic three-dimensional architectures of native pancreas are retained after decellularization.The extracellular matrix (ECM) ultrastructures were well preserved.Immunofluorescence staining showed that collagen Ⅰ,Ⅳ,fibronectin and laminin were maintained in the APB after decellularization.DNA quantification of APB decreased significantly (P ＜ 0.05).The histological analysis of the subcutaneous implantation site showed the presence of immunological response surrounding the partially degraded APB.The histological remold score was 10.4 ± 1.8 at 14 days and 13.8 ± 1.3 at 28 days.The proliferation of AR42J cell line with the expression of amylase as the marker of exocrine acinar cells can be detected on the recellularized APB.Conclusion APB in this study met the stringent requirement to define a successful decellularization.The technique allowed the efficient generation of APB with preserved 3D architecture and represented a biocompatible scaffold capable of integrating within host tissue.

Objective To investigate the curative effect of neural stem cells (NSCs) transplantation on sequela after traumatic intracranial hematoma. Methods 20 patients with sequela after traumatic intracranial hematoma were treated with NSCs transplantation. Cells were engrafted into subarachnoid cavity via lumbar puncture. They were assessed with Functional Independence Measure (FIM) before and half a year after the transplantation. Results The FIM scores were significantly increased after the transplantation (P<0.01).Conclusion NSCs transplantation could promote functional recovery and improve the living quality of patients with sequela after traumatic intracranial hematoma in the aspects of self-care, sphincter control, mobility, locomotion, communication and social adjustment/cooperation.

Objective: To compare the sequence diversity of HVR1 in the putative envelope protein E2 of the genotypeⅡand genotype Ⅲ HCV in Chinese. Methods: The cDNAs[nucleotide(nt)1449-1586(HCV-J) or nt1460-1582(HCV-J6)] derived from plasma of 55 patients infected with genotype Ⅱ HCV and 38 patients infected with genotype Ⅲ HCV were amplified,purified and directly sequenced by RT-nested polymerase chain reaction(PCR) and dideoxynucleotide chain termination method. Results: The HVR1 was found in amino acid(aa) 384-408 positions of both types HCV E2 protein. There were 5 similar conserved amino acids in 2 types HCV HVR1:aa385(Thr), aa389, 390, 406(Gly)and aa403(Phe).Besides, 401(Ser) was also highly conserved in genotype Ⅱ HCV HVR1. Although the variation characteristic of 2 types was similar, but the sequence diversity(SD),the kinds and frequency of some amino acids in some HVR1 positions and the conserved region near the HVR1 had some differences between 2 genotypes. Conclusion: Further study on the diversity of HVR1 and its biological significance will be helpful to understand the mechanism of HCV persistent infection and the development of HCV vaccine.

Objective To explore whether hypoxia-inducible factor-1α( HIF-1α )is involved in oxygenglucose deprivation (OGD)induced caspase-independent cell death in primary cortical neurons and whether their expression is infected by necrostatin-1 ( Nec-1 ).Methods ( 1 ) Primary cerebrocortical neurons were cultured for 14 days.Pretred z-VAD.Fmk (z-VAD)and Nec-1 with 0.1,1,5,10,25 and 50 μ mol/L before the neurons were exposed to OGD for 2 hours and reoxygenated for 12 hours,then cell viability was determined by measure LDH level.(2)Pretred z-VAD before the neurons were exposed to OGD for 2 hours,then reoxygenated for 0,2,6,12,24and 48 hours.Then western blot analysis protein level of HIF-1 α ;rt-PCR check its RNA level.(3)Pretred z-VAD and Nec-1 with 25μ mol/L before the neurons were exposed to OGD for 2 hours and reoxygenated for 12 hours.Then western blot analysis protein level of HIF-1 α; rt-PCR check its RNA level.Result ( 1 ) When cells were pretread Nec-1 with 5 μ mol/L(6.97 ± 0.06),the level of LDH was lower than cells untreated( 14.23 ± 0.08 ) (P＜ 0.05);At 25 μmol/L( 2.21 ± 0.05),the level of LDH was essentially the same as that of the control( 1.03 ±0.03 ) (P＞0.05).(2)The protein level of HIF-1 αwas different from normal (0.24 ±0.01 ) when exposed to OGD for 2 hours and reoxygenated for 2 hours (0.57 ± 0.09) and was highest after cells were exposed to OGD for 2 hours and reoxygenated for 12 hours(0.91 ± 0.08 ) (P＜ 0.05 ).The RNA level of HIF-1 α when cells were exposed to OGD was not deferent from normal (P ＞ 0.05 ).( 3 ) When cells were pretread with Nec-1 (0.32 ± 0.04 ),the protein level of HIF-1α were lower than untreated(0.83 ±0.03) (P＜0.05),but the RNA level of HIF-1α had no deference(P ＞ 0.05).Conclusion HIF-1α was involved in cell' s caspase-independent cell death;Nec-1 can protect neurons through inhibiting the expression of HIF-1α.

Objective To investigate the effects of hyperbaric Oxygen (HBO)exposure on Caspase-3 and Bcl-2 mRNA ex-pressions in both internal carotid artery (ICA)and basal artery (BA)in rabbits.Methods Twenty-four healthy adult New Zealand rabbits were randomly divided into 2 groups:HBO group and the control group,with each group consisted of 12 animals.The rab-bits in the HBO group were exposed to HBO at 2.2 ATA for 60 minutes each day for 3 successive days.The rabbits in the control group were normally fed without any treatment.Real-time PCR was used to detect Caspase-3 and Bcl-2 mRNA expressions in both ICA and BA in 2 groups.Results HBO significantly decreased the Caspase-3 mRNA expression [(0.038 ±0.006 )vs .(1.000 ± 0.225)]and increased the Bcl-2 mRNA expression [(1.877 ±0.1 69)vs .(1.000 ±0.364)].In ICA,HBO similarly decreased the Caspase-3 mRNA expression [(0.41 9±0.091)vs .(1.000 ±0.1 75)]and increased the Bcl-2 mRNA expression [(1.269 ±0.270) vs .(1.000±0.1 1 7)]in BA.All the differences mentioned above were of statistical significance (P <0.01).Conclusion HBO ex-erts an inhibition effect on apoptosis in cerebral vascular endothelial cells.The mechanism may be related to inhibiting the expres-sion of Caspase-3 mRNA and promoting the expression of Bcl-2 mRNA.