Objective: To investigate immune-regulating effects of Ar-He targeted cryoablation for the treatment of the advanced non small cell lung cancer (NSCLC). Methods: One hundred NSCLC patients were enrolled in this study. The serum levels of IL-2, TNF-α and IL-12 were measured by enzyme-linked immunosorbent assay (ELISA) before and 10 days after Ar-He targeted cryoablation. Low-density mononuclear cells (NINC) were separated by Ficoll-Hypaque as effector cells. Human lung cancer SPC-A1 and K562 cells were selected as target cells. The activities of natural killer (NK) cells and cytotoxic T-lymphocytes (CTL) were determined by MTT assay. Results: Of the 100 patients, 3 cases achieved complete remission (CR), 34 cases achieved > 50% remission (PR), 47 cases had stable disease (SD), and 16 cases had progressive disease (PD). The one-year survival rate was 43% and the median survival time to progression was 4 months. The non specific tumor-killing ability of NK cells increased by 83% and the specific tumor-killing activity of CTL increased by 67%. The serum levels of IL-2, TNF-α, and IL-12 were elevated significantly. The positive expression of CD3+ and CD4+ on T cells and the ratio of CD4+/CD8+ also increased. Conclusion: The effectiveness of Ar-He targeted cryoablation for the local treatment of NSCLC could be observed by imaging analysis. Ar-He targeted cryoablation regulates and stimulates self immunological function of NSCLC patients. Ar-He targeted cryoablation provides an effective method especially for old and surgery nontolerant patients.

Objective To Explore the effect of Cassia Seed budding transforming organic vanadium. Methods Separate organic vanadium from inorganic vanadium in Cassia Seed sprouts by dislysis bag. The content of total vanadium and inorganic vanadium were determined by using the method of Graphite Atomic Absorption Spectrophotometry. The transformation efficiency of organic vanadium in the Cassia Seed spouts was calculated. Results The seed coat of Cassia Seed could accumulate massive inorganic vanadium existing in environment during the process of its natural growing period,but the transformation efficiency of organic vanadium mainly occurred inside seed coat of Cassia Seed. Condusion Cassia Seed budding is one pathway of the biologic organification of vanadium.

Humans ; Mining ; Noise, Occupational ; statistics & numerical data ; Occupational Exposure ; statistics & numerical data ; Work Schedule Tolerance

Objective To study the relationships between hepatocellular carcinoma (HCC) and the polymorphisms,promoter methylation,and expression of glutathione S-transferases P1 gene (GST) P1 gene.Methods Using methylation-special PCR (MSP),the methylated status of CpG islands of GSTP1 gene in tumor tissues of 53 HCC and its adjacent nontumor tissues were studied.The enzyme activities of GSTP1 were evaluated by ultraviolet colormetry.And using PCR-RFLP,the genetic polymorphisms of the GSTP1 genes of 74 healthy controls and 53 HCC patients were studied.Results The diffe-rences of the frequency of GSTP1 Ile/Ile,Ile/Val and Val/Val genotypes between HCC patients and the normal controls did not reach statistical significance (X~2=0.84,v=2,P=0.656).The frequency of methyla- tion of CpG islands of GSTP1 gene was significantly higher among the HCC tumor tissues when com- pared to the corresponding nontumor tissues (X~2=19.08,P＜0.001),and significantly higher in stageⅢ-Ⅳcases when compared to the stageⅠ-Ⅱcases (X~2=4.84,P=0.028).GSTP1 enzyme activities of cytoplasm in tumor cells were lower significantly than that in the adjacent nontumor tissues (t=2.49, P=0.014),and significantly higher in stageⅠ-Ⅱcases when compared to the stageⅢ-Ⅳcases (t= 2.31,P=0.025).On the other hand,the GSTP1 enzyme activities of cytoplasm in tumor cells with methylated status of GSTP1 gene were significantly lower than that in tumor cells with unmethylation (t=3.50,P=0.001).Conclusion GSTP1 inactivation via CpG island hypermethylation may contrib- ute to the pathogenesis of HCC.

Objective To explore the different influence of antiepileptic drugs(AEDs)at therapeutic levels to the maturation of brain.Methods 180 healthy Sprague-Dawley(SD)rats were divided into infant and adult group.Each age group was administered with PB,CNP,VPA,TPM or normal saline respectively in persistent 5 weeks.The steady-state plasma concentrations of AEDs at the experimental dosage were coincided with the range of clinical therapeutic concentrations.After AEDs withdrawed,the effects of AEDs on cognitive function were assessed by Morris water maze and two-way shuttle box at different time points.Body and brain weight were got immediately when the rats were sacrificed.Histological changes of brain were observed by HE staining,Nissl staining and transmission electron microscopy.Results(1) For immature rats,1 day or 14 days after AEDs withdrawed,there were significant differences between groups exposed to PB or CNP and control group in escape response latency(ERL)in the two-way shuttle box.Even after one month ERLs of immature rats receiving CNP((6.05?2.04)s)or PB((5.81? 1.75)s)were still longer than that of untreated controls((4.75?2.43)s,P

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 explore the influence of intestinal trefoil factor (ITF) on interleukin-6(IL-6) in neonatal rat with necrotizing enterocolitis(NEC) ,and to discuss the protective machanism of ITF on NEC.Methods Thirty-two neonatal rats were divided randomly into four groups,group A as control group,group B as NEC group,group C as NEC+NS 0.2 mL group,group D as NEC+ITF 0.2 mg group.NEC model of neonatal rats were established.On the 4th day,all the subjects were put to death.Intestinal tissue within the boundary of ileum and cecum was obtained to observe histological changes.Other intestinal tissue was treated into homogenate.After the homogenate was centrifuged,supernates were used to test the density of IL-6.Results The density of IL-6 significantly decreased in group A,D than those in group B and C (Pa0.05).The pathological lesions indicated that intestinal tissue necrosis was severe in group B and C,which was graded as 3 points,but obviously lessen in group D,which was graded as 1 point,with ITF interfering.Conclusions Intestinal inflammation is ameliorated after ITF are injected hypodermically or intraperitoneally.ITF may provide a brand-new way for the therapy of NEC in neonatal rats.

Objective To study the differences of chromosomal structure among Yersinia pestis strains isolated from China,and to investigate the reasons of chromosomal rearrangement events occurred in Yersinia pestis as well as the possibility of strain identification and phylogenetic analysis based on the chromosomal rearrangement features.Methods According to the genome sequence data downloaded from web of National Center for Biotechnology Information (http://www.ncbi.nlm.nih.gov/genome),alignment of all the coding sequences (CDSs) among five strains(American strain CO92 as reference and other four completely sequenced strains from Inner Mongolia,Jianchuan of Yunnan,Yulong of Yunnan,Naqu of Tibet in China named 91001,D182038,D106004 and Z176003 as comparison strains) was performed,and then the chromosome of Yersinia pestis was divided into several large DNA segments (named chromosomal plate in the text) according to the similarity of CDSs.Plate arrangement patterns in each strain' s chromosome and gene content of breakpoint regions were determined.Finally,genetic relationships among Yersinia pestis strains were analyzed on the basis of rearrangement diversity from paired-comparison.Results Yersinia pestis chromosomes of strains CO92,D182038,D106004,91001 were composed of 44 relatively independent plates,except strain Z176003.Gene order was very stable within each plate,while it was movable between the plates.Comparing with the reference strain CO92,13 rearrangement events occurred in the chromosomes of both strain D182038 and strain D106004,and 14 rearrangement events involved in Z176003,while 37 rearrangement events occurred in 91001.Paired-comparison data showed that only 8 plates order differences were existed between D106004 and Z176003.Forty-three breakpoint regions were identified on the chromosome of strain CO92,and 39 of them contained insertion sequences,and 25 of them were IS100.Conclusions Yersinia pestis genome represents a high degree of genetic flux,and chromosomal structures of strains are significantly different from each other.Chromosomal rearrangement events is closely related to the large number of insertion sequences in the Yersinia pestis chromosome.Rearrangement diversity among Yersinia pestis strains could reflect their genetic relationships.