AIM: (1) To investigate the effect of amniotic membrane transplantation (AMT) on rabbit conjunctival surface reconstruction with severe alkali burns. (2) To evaluate the possibility of AMT treatment for ocular alkali burns during recovering stage.METHODS: Animal models were established on 30 eyes of rabbits by creating severe alkali burns on the conjunctiva from the upper corneal limbus to the upper conjunctival fornix.Preserved human amniotic membrane transplantations and reconstruction of conjunctival fornix were performed at one week after injury (recovering stage). Epithelium growth of burned area after transplantation was observed using light microscope at 1, 2, 3, 4, and 8 weeks. Conjunctival tissue in transplantation area was collected at 1, 4 and 8 weeks. The ultrastructure of the collected tissue was studied by electron microscope. The results were compared with control group,which received only vitamin C subconjunctival injection and antibiotic eye drops as treatment for alkali burn. Exterior eye pictures were also taken at the end of the observation, the width from upper corneal limbus to the edge of upper fornix was measured. Data was analyzed statistically.RESULTS: 1) Tn the transplant group, conjunctival epithelium growth was observed in the area of AMT under both light and electron microscope 1 week after surgery. At 4weeks, conjunctival epithelium with goblet cells that resembled normal conjunctival tissues was observed in the whole amniotic membrane area. At 12 weeks, the conjunctival epithelium on the amniotic membrane was well formed, and the connective tissue under the epithelium was loose at the fornix. No fibrosis was identified. In contrast, conjunctival epithelium necrosis was observed in the control group at 2weeks after alkali burns. Re-epithelization did not occur through the 12-week observation. Severe fibrosis with inflammatory cells infiltration was observed between 4 to 8weeks. At 12 weeks, fibrosis of the connective tissue at the fornix developed and there were no conjunctival epithelium covering the burned area. 2) In the transplant group, the conjunctiva in transplanted area had no scarring and appeared smooth at 12 weeks. Upper fornix was reconstructed. The depth of fornix was 7.9±0.3mm (7.6-8.2mm), which was approximate to the normal depth 8.2±0.2mm (8.0-8.4 mm,P＞.05). While in the control group, the burned area appeared rough with granuloma formation and severe scarring. Upper fornix became shallow. The depth of fornix was 3.1±1.7mm(1.0 to 4.5mm.), and significant difference was found between control and transplant group (P＜0.01).CONCLUSION: Human amniotic membrane preserved in glycerin can promote cell adhering, migrating and differentiating of normal conjunctival epithelium.Reconstruction of conjunctival surface in early stage of alkali burn can be achieved by AMT. AMT can effectively prevent symblepharon formation.

Objective To investigate the methods and clinical significance of breast cancer treated with photodynamic.Methods From June to December in 2005,photodynamic therapy was used in 12 cases confirmed intramammary lymph node metastasis before operation and 15 cases confirmed chest wall recur- rences by means of lymph node imaging.Results The intramammary lymph node metastasis whose diameter between 0.5～1.0cm measured by lymph node imaging preoperatively completely disappeared when rechecked 3 months postoperatively.Chest wall recurrence regions of breast cancer whose diameter less than 1.0 cm completely remitted.Conclusion Photodynamic therapy is helpful to eliminate the intramammary lymph node metastasis and to cure the postoperative chest wall recurrence of breast cancer.

Objective To provide the theoretical basis for the application of cortical somatosensory evoked potential (CSEP) in monitoring the function of the spinal cord to prevent postoperative neurological dysfunction. Methods Thirty-three New Zealand rabbits were randomly divided into 6 groups: 8 were chosen as control group to eliminate the influence of anesthesia and surgery on the evoked potential; the other 25 were assigned to 5 sub-experimental groups (n=5) according to the artery number being ligatured in the left renal arteries and the spinal arteries. Baseline evoked potential in each group was noted immediately after anesthesia; the CSEP were recorded at different time points (before vascular ligation, 30 min and 2 d after vascular ligation). Motor functions were assessed after narcotic conscious and 2 d after vascular ligation. The specimens were taken for HE staining. Results The latency was not sensitive to spinal cord ischemia and no significant difference of that was found between the experimental groups and the control group (P>0.05); except that, the changes of theamplitudes were very complex and the specificity of motor function was decreased. The amplitude reduced and then gradually restored in the 2, 3 and 4 levels of ligation. The changes of amplitude could indicate the degree of pathological damage in the spinal cord and its motor function. Conclusion Complex amplitude of somatosensory evoked potential can be found in the acute phase of ischemia in the spinal cord. Specificity of motor function is poor resulting from its signal averaging process. Motor evoked potential monitoring in the operation should also be added in the detection of the spinal cord.

Aim To study the effect of Guben Yanling pills on liver aging in aging mice and the related mech-anism.Methods The mice were randomly divided in-to blank control group,model group,vitamin E group(0.1 g·kg-1)and low,medium and high dose groups(0.59,1.17,2.34 g·kg-1)of Guben Yan-ling pills.The aging mouse model was established by subcutaneous injection of D-galactose(150 mg·kg-1)into the back of neck.At the same time of mod-eling,the corresponding drugs were given by gavage once a day for six weeks.The main organ indexes were calculated.HE staining was used to observe the mor-phology of liver tissue.Colorimetry was used to detect the activity of β-galactosidase in liver.ELISA was used to detect the content of TNF-α,IL-1 β,IL-6,IL-4,IL-10.Western blot was used to detect the protein relative expression level of IKKβ,Iκ Bα,NF-κB p65.Immunofluorescence was used to detect the expression level of NF-κB p65.Results Compared with the blank control group,the organ index of the brain,liv-er,kidney,spleen,and thymus in the model group decreased(P＜0.05,P＜0.01),the activity of β-galactosidase increased(P＜0.01),liver tissue mor-phology and structure were significantly damaged,the content of TNF-α,IL-1 β and IL-6 increased(P＜0.01),the content of IL-4 and IL-10 decreased(P＜0.01),the levels of IKKβ,NF-κB p65 in-creased(P＜0.01),the levels of IKBα decreased(P＜0.01),and the levels of NF-κB p65 in nucleus increased(P＜0.01).Compared with the model group,the organ indexes of brain,liver,kidney,spleen,and thymus in each dose group of Guben Yan-ling pills increased(P＜0.05,P＜0.01),the activity of β-galactosidase decreased(P＜0.01),the morpho-logical and structural damage of liver tissue was signifi-cantly improved,the content of TNF-α,IL-1 β and IL-6 decreased(P＜0.01),the content of IL-4 and IL-10 increased(P＜0.01),the levels of IKKβ,NF-κB p65 decreased(P＜0.01),the levels of IκBα in-creased(P＜0.01),and the levels of NF-κB p65 in nucleus decreased(P＜0.01).Conclusions Guben Yanling pills can antagonize liver aging in mice,and its mechanism may be related to inhibiting the activa-tion of NF-κB signaling pathway in liver,downregulat-ing downstream pro-inflammatory factor levels,upregu-lating anti-inflammatory factor levels,and alleviating inflammation in liver.

BACKGROUNDTo investigate the differential expression levels of thymosin beta 10 (T beta 10) and the corresponding changes of actin filament organization in human tumor cell lines with different metastatic potential.

METHODSFour groups of nine human tumor cell lines with different metastatic potential were analyzed for the amount of T beta 10 mRNAs by Northern blot and for their peptide expression levels by immunohistochemistry. The filamentous actin (F-actin) was observed by staining of TRITC-phalloidin to detect changes in actin organization.

RESULTSIn comparison with non-/weakly metastatic counterparts, T beta 10 was upregulated in highly metastatic human lung cancer, malignant melanoma and breast cancer cell lines. Staining of TRITC-phalloidin revealed less actin bundles, a fuzzy network of shorter filaments and some F-actin aggregates in the highly metastatic tumor cells. Meanwhile, the actin filaments were robust and orderly arranged in the non-/weakly metastatic cancer cell lines.

CONCLUSIONT beta 10 levels correlate positively with the metastatic capacity in human tumors currently examined. The increasing metastatic potential of tumor cells is accompanied by a loss of F-actin, poorly arranged actin skeleton organizations and presence of F-actin aggregates. There is a consistent correlation between the elevated T beta 10 expression and the disrupted actin skeleton.

Actin Cytoskeleton ; ultrastructure ; Blotting, Northern ; Cell Line, Tumor ; Humans ; Immunohistochemistry ; Neoplasm Metastasis ; genetics ; ultrastructure ; RNA, Messenger ; analysis ; Thymosin ; analysis

OBJECTIVETo investigate the protective effects of the natural medicinal monomer isopsoralen (ISR) with estrogenic activity against oxidative damage in human lens epithelial cells B3 (HLE-B3) caused by hydrogen peroxide (H(2)O(2)) and to pursue the possible mitochondrial proteomic regularity of the protective effects.

METHODSHLE-B3 cells were treated with H(2)O(2) (300 μ mol/L), β-estradiol (E(2): 10(-8) mol/L) and H(2)O(2), ISR (10(-5) mol/L) and H(2)O(2), or left untreated. Altered expressions of all mitochondrial proteins were analyzed by protein array and surfaceenhanced laser desorption ionization time of flight mass spectrometry (SELDI-TOF-MS). The mass/charge (m/z) ratios of each peak were tested by the Kruskal-Wallis rank sum test, and the protein peak value of the m/z ratio for each treatment by pair comparison was analyzed with the Nemenyi test.

RESULTSH(2)O(2) up-regulated the expressions of two protein spots (with m/z of 6532 and 6809). E(2) mitigated the oxidative damage, and the expression of one protein spot (m/z 6532) was down-regulated. In contrast, ISR down-regulated both of protein spots (m/z 6532 and 6809).

CONCLUSIONSISR could effectively inhibit H(2)O(2)-induced oxidative damage in HLE-B3 cells. The protein spot at m/z of 6532 might be the target spot of ISR against oxidative damage induced by H(2)O(2).

Cell Line ; Epithelial Cells ; drug effects ; metabolism ; pathology ; Estradiol ; pharmacology ; Furocoumarins ; pharmacology ; Humans ; Hydrogen Peroxide ; toxicity ; Lens, Crystalline ; pathology ; Mitochondria ; metabolism ; Oxidation-Reduction ; drug effects ; Oxidative Stress ; drug effects ; Protective Agents ; pharmacology ; Proteome ; metabolism ; Proteomics ; methods

OBJECTIVETo study HIV, HBV and HCV infections in intravenous drug users.

METHODS2025 blood samples from intravenous drug users were collected from Sichuan, Hunan, Guangxi and Xinjiang regions, and tested for anti-HIV, anti-HCV, HBsAg using enzyme-linked immuno-sobent assays (ELISAs).

RESULTSThe positive rates of anti-HIV,anti-HCV and HBsAg were14.7%-30.4%, 60.7%-85.5% and 6.6%-22.4% in the intravenous drug users, respectively. The co-infection rates of HIV/HBV, HIV/HCV, HCV/HBV and HIV/HCV/HBV were 0%-0.4%, 11.6%-27.2%, 2.3%-14.3% and 1.6%-4.8% respectively in this population.

CONCLUSIONThe infection rates of HIV, HBV and HCV were higher in the intravenous drug users than that in general populations in the same regions, and HIV/HCV co-infection appeared most frequent in this population.

China ; epidemiology ; HIV Infections ; epidemiology ; Hepatitis B ; epidemiology ; Hepatitis C ; epidemiology ; Humans ; Prevalence ; Substance Abuse, Intravenous

The essence of endogenous turbidity in Chinese medicine (CM) is different from cream, fat, phlegm, retention, damp, toxicity, and stasis. Along with the development of modern scientific technologies and biology, researches on the essence of endogenous turbidity should keep pace with the time. Its material bases should be defined and new connotation endowed at the microscopic level. The essence of turbidity lies in abnormal functions of zang-fu organs. Sugar, fat, protein, and other nutrient substances cannot be properly decomposed, but into semi-finished products or intermediate metabolites. They are inactive and cannot participate in normal material syntheses and decomposition. They cannot be transformed to energy metabolism, but also cannot be synthesized as executive functioning of active proteins. If they cannot be degraded by autophagy-lysosome or ubiquitin-prosome into glucose, fatty acids, amino acids, and other basic nutrients to be used again, they will accumulate inside the human body and become endogenous turbidity. Therefore, endogenous turbidity is different from final metabolites such as urea, carbon dioxide, etc., which can transform vital qi. How to improve the function of zang-fu organs, enhance its degradation by autophagy-lysosome or ubiquitin-prosome is of great significance in normal operating of zang-fu organs and preventing the emergence and progress of related diseases.
Autophagy ; Humans ; Medicine, Chinese Traditional ; Proteasome Endopeptidase Complex

OBJECTIVETo develop an in vitro assay that allows the culture and identification of a single human bone marrow progenitor closely related to hematopoietic stem cell, which is more primitive than LTC-IC, and to find an efficient culture conditions for NK-IC expansion.

METHODSFusion protein IL6/IL2 was reconstructed and expressed in E. coli DH5 alpha. ML-IC was determined by watching if the single cell can give rise to secondary progenitors with both LTC-IC and NK-IC characteristics. LTC-IC frequency was determined by the CFC clonogenic methylcellulose assay. NK-IC frequency was determined by phenotyping CD56 positive NK cells. The effect of FPIL6/IL2 on the expansion of NK-IC was examined by comparing the colony number of NK cells before and after the culture.

RESULTSAfter the initial 4-week expansion culture, we showed that (25.75 +/- 5.68)% of freshly sorted Lin-/34+/DRdim cells were able to generate functional NK-IC in one or more secondary FPIL6/IL2 cultures, whereas (6.81 +/- 1.97)% in the control. A total of 102 NK-IC cells were present when were cultured for 6-7 weeks in FPIL6/IL2 expansion medium, which was much higher than the 33 NK-IC cells in the control.

CONCLUSIONML-IC assay will prove useful to assess a very primitive hematopoietic cell with multilineage generative capacity. FPIL6/IL2 is capable of initiating and promoting NK-IC expansion greatly in ex vivo cultures in terms of net-conservation and net proliferation.

Animals ; Biological Assay ; Cell Culture Techniques ; methods ; Cell Differentiation ; Cell Division ; Cells, Cultured ; Coculture Techniques ; methods ; Hematopoietic Stem Cells ; cytology ; Humans ; Interleukin-2 ; biosynthesis ; genetics ; pharmacology ; Interleukin-6 ; biosynthesis ; genetics ; pharmacology ; Killer Cells, Natural ; cytology ; metabolism ; Mice ; Recombinant Fusion Proteins ; pharmacology ; Stromal Cells ; cytology

OBJECTIVETo investigate the relationship between cyclin D2 and P210(BCR/ABL) tyrosine kinase in chronic myelogenous leukemia (CML).

METHODSRT-PCR, Western blot and flow cytometry were performed to detect the expression of cyclin D2 in K562 cells and in K562-ib-eGFP cells which express intracellular single-chain antibody (sFv, intrabody) against ABL tyrosine kinase domain.

RESULTSCyclin D2 expression in K562-ib-eGFP cells was 18.90% which was lower than that of control K562 cells (48.10%), and the number of S-phase cells in K562-ib-eGFP was 40.40% which was much lower than that in K562 cells (64.34%).

CONCLUSIONCyclin D2 is a potential down-stream signal molecule of the p210(BCR/ABL) tyrosine kinase in CML. The altered expression of cyclin D2 may contribute to the over proliferation of CML cells.

Blotting, Western ; Cyclin D2 ; Cyclins ; analysis ; genetics ; Flow Cytometry ; Genes, abl ; Humans ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction