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.

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

OBJECTIVETo perform the detection of spermatozoal gene expression in order to accelerate the study of spermatozoal molecular biology.

METHODSTo collect the healthy adults sperm and lymphocytes respectively, and then to extract the total RNAs from them by RNeasy mini kit (QIAGEN) or Trizol reagent. Corresponding cDNAs were produced, digested, ligated, finally labeled with Cy3 (sperm) and CyS (lymphocyte) in the course of RD amplifying reactions. Hybridization with self-made microarrays contained 560 probes was carried out after the labeled cDNAs pured by PCR Product Purification Kit.

RESULTSAmong the 560 probes, 72 genes were up-regulated, 321 genes were down-regulated, the others had no different expression. Furthermore, genes associated with replication, transcription, translation and regulative functions were non-different expression or down-regulated, and those belonged to the spermatogenesis associated, sperm associated antigen were up-regulated, but those involved in the glycolysis were up-regulated, in the oxidative phosphorylation were down-regulated.

CONCLUSIONIt had successfully confirmed that there were a plenty of genes expressed in sperm, furthermore the genes expressed were accorded to spermatozoal functions and characteristics.

Adult ; Down-Regulation ; Gene Expression Profiling ; Gene Expression Regulation ; Humans ; Lymphocytes ; metabolism ; Male ; Middle Aged ; Oligonucleotide Array Sequence Analysis ; Polymerase Chain Reaction ; RNA ; isolation & purification ; Spermatozoa ; metabolism ; Up-Regulation

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

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 determine the efficacy of phosphocreatine kinase in the early diagnosis of compartment syndrome.

METHODSForty patients with compartment syndrome of limbs were reviewed from 2005 to 2008 including 34 males and 6 females with an average age of (37.03 +/- 13.02) years. Monitoring phosphocreatine kinase continuously and dynamically after injured 2, 24 hours, 1, 2, 3 and 4 weeks later. The concentration of CK were measured by using Japanese Olympus automatic biochemistry analysator. The muscle preparations from affected extremity were taken after operation and 1, 2, 3 weeks later for biopsy.

RESULTSTwo hours later after injury, the contents of CK increased sharply and the contents of CK were about 20 times more than the nomal. Twenty-four hours later, the contents of CK reached its maximum,the contents of CK were about 42 times more than the nomal. One week later, the contents of CK recovered to normal level. Pathological changes of muscle were irreversible.

CONCLUSIONThe change of the contents of CK can reflect the progression of disease objectively. If it increased sharply, the chance of compartment syndrome was high. Monitored it dynamicly and continuously can provide assistant for early diagnosis of compartment syndrome and evaluating pathogenetic condition.

Adolescent ; Adult ; Aged ; Child ; Compartment Syndromes ; blood ; diagnosis ; Creatine Kinase ; blood ; Female ; Humans ; Male ; Middle Aged ; Time Factors

OBJECTIVETo investigate the expression of thymosin beta10 (Tbeta10) and related 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 expression of Tbeta10 mRNA detected by northern-blot and its peptide by immunohistochemical staining. The filamentous actin (F-actin) was stained with TRITC-phalloidin to detect changes in actin organization.

RESULTSIn comparison with the non and/or weakly metastatic counterparts, Tbeta10 was upregulated in highly metastatic human lung cancer, malignant melanoma and breast cancer cell lines. TRITC-phalloidin staining revealed less actin bundles and a fuzzy network of shorter filaments in the highly metastatic tumor cells, while in the non and/or weakly metastatic cancer cell lines, there were thick and orderly arranged actin filaments.

CONCLUSIONSTbeta10 levels correlate positively with the metastatic phenotype in human tumors currently examined. The increased metastatic potential of tumor cells is accompanied by the loss of F-actin and poorly organized actin skeleton. There is a consistent correlation between the elevated Tbeta10 expression and the disrupted actin skeleton.

Actins ; analysis ; Blotting, Northern ; Cell Line, Tumor ; Humans ; Immunohistochemistry ; Neoplasm Metastasis ; Thymosin ; analysis