[Endothelium Aging and Oxidative Stress].

In the endothelium, ROS mainly derive from mitochondria, endothelial nitric oxide synthases and NADPH oxidases 4. Excessive ROS are a major cause of oxidative stress, the primary stimulus of vascular dysfunction and oxidative stress-related diseases. However, cellular evolution has made possible the development of adaptive antioxidant systems that scavenge excessive ROS, such as Nrf2/Keapl-ARE, PPAR-y, SIRT and FOXO, etc. Among them, the Nrf2/Keapl-ARE signaling pathway is perhaps the most prominent. What is more, there are the "crosstalk" among these antioxidant stress-related signaling pathways aim to alleviate oxidative stress injurys and promote cells survival. The understanding of the relationship between endothelial aging and oxidative stress may serve as a therapeutic clues in the treatment of oxidative stress-related diseases.

[Affection of acupotomy lysis on leu-enkephalin (L-ENK) content in different parts of centrum of rats with knee osteoarthritis].

OBJECTIVE: To study the analgesia mechanism of needle-knife lysis in spinal cord and other parts of central nervous system by comparing the changes of Leu-Enkephalin (L-ENK) content in different parts of centrium of rats undergone needle-knife lysis and electro-acupuncture respectively. METHODS: Sixty healthy SD rats were randomly devided into normal control group, model group, needle-knife lysis (NKL) group and electro-acupuncture (EA) group. 4% papain solution mixed with 0.3 mol/L cysteine solutin in the ratio of 1:1, paused for 0.5 h,injected the mixture, 20 microl each time,into the left knee joint cavities of rats in model, NKL, EA groups at the 1st, 4th, 7th day. After 4 weeks in NKL group and EA group were treated with needle-knife lysis and electro-acupuncture, respectively. Three weeks after treatment, samples of spinal cord of the swollen part of rat waists and rat brains were taken from and the content of L-ENK of medulla oblongata, midbrain, pituitary gland, and hippocampus were measured. RESULTS: L-ENK content of model group increased higher than that of normal control group in spinal cord, hippocampus and midbrain (P < 0.01); there were no significant difference between normal control group and modle group on L-ENK in medulla oblongata and thalamus (P > 0.05). After intervening of NKL or EA, L-ENK content of NKL group increased higher in hippocampus than that of model group and EC group (P < 0.05); but L-ENK content of NKL group in midbrain was lower than that of model group (P < 0.05). CONCLUSION: Needle-knife lysis has characteristic of regulation for the L-ENK content in different parts of central nervous system of rats with knee osteoarthritis, and analgesic effect of needle-knife was possibly related with regulation of center L-ENK.

[The significance of serum GM and BG antigens assay for invasive fungal infections in hematological malignancies patients].

OBJECTIVE: To evaluate the diagnostic value of serum galactomannan antigen (GM) and (1→3)-ß-D-glucan antigen (BG) assay in invasive fungal infections (IFI) in the patients with hematologic malignancies and the role in monitoring therapeutic response. METHODS: Fifty one patients with hematological malignancies met the criteria for inclusion: (1) body temperature above 38°C for 48 hours, (2) failure to respond to broad-spectrum antibiotic treatment, or (3) temperature rose again after the responded drop. Blood samples were collected twice at the first week, then once a week in at least four weeks. The double antibody sandwich enzyme-linked immunosorbent assay (ELISA) and colorimetric assay were used for detecting GM and BG. The positive GM test is defined as two consecutive tests at different time GM value > 0.5 or > 0.8 and the positive G test is defined as BG value > 80 pg/ml. The patients were assigned into four groups as proven, probable, possible, and non-fungal infection respectively, and 21 normal volunteers were as controls. RESULTS: Two hundred and forty serum samples were collected from 51 patients including 2 of proven IFI, 26 probable IFI, 17 possible IFI and 6 non-fungal infection. The true-positive group including the proven and probable groups, and true negative group was the non-fungal infection group. GM tests were positive in 21 of 28 cases in true positive group, and only one of 6 cases in non-fungal infection. The sensitivity, specificity, positive predictive value and negative predictive value were 75%, 83.3%, 95.5% and 41.7%, respectively. G tests were positive in all 28 cases of the true positive group, and 4 in 6 non-fungal infection cases. The sensitivity, specificity, positive predictive value and negative predictive value were 100%, 33.3%, 87.5% and 100%, respectively. G test is more sensitive than GM test (P = 0.015), but there was no significant difference in specificity of the two tests (P = 0.242). In 19 of 21 patients with GM test positive, anti-fungal treatment was effective, and GM value gradually decreased to negative, two invalid patients were persistent with GM test positive. After two weeks treatment, the average GM value was significantly lower in the effective group than in the ineffective group (P < 0.05). BG values in the responded patients showed a gradual decline similar to that of GM values, but not to negative. The changes of BG value in ineffective group varied with a trend upward. The changes in BG value had no relation with treatment effectiveness. CONCLUSIONS: Serum GM and BG antigens detection provides strong evidence for early diagnosis of IFI. Combination of GM and G tests can improve the diagnostic specificity and reduce the false positive GM test seems superior to G test for monitoring GM and BG values during treatment.

[Toxic effect of immunosuppression agent Everolimus on insuloma cells and pancreatic islets of SD rat].

OBJECTIVE: To investigate the effect of immunosuppression agent Everolimus on the viability and function of insuloma cells (INS-1) and pancreatic islet cells cultured in vitro. METHODS: INS-1 cells and islets were treated with a series of concentrations of immunosuppression agents (Everolimus, Cyclosporin A, Sirolimus and Mycophenolate Mofetil). The viability of INS-1 cells and rat pancreatic islets were determined with MTT and the function of INS-1 cells and rat pancreatic islets was evaluated with Glucose-stimulated insulin secretion assay. RESULTS: Above the clinical blood concentration, the inhibition rate of islet cell proliferation in the high concentration group of Everolimus and Sirolimus was significantly lower than that of Cyclosporin A and Mycophenolate Mofetil group (P < 0.05); Everolimus in the blood drug level, like other immunosuppressive agents, can inhibit the function of insulin secretion, and the stimulation index of each group was no significant difference. CONCLUSION: Compared to Mycophenolate Mofetil and Cyclosporin A, Everilimus and Sirolimus demonstrate lower toxicity effect on INS-1 cells and rat pancreatic islets in vitro and Everolimus is expected as a new type of immunosuppressive agent used in clinical islet transplantation.

[Debating some issues on the formulation and application of standardized manipulations of acupuncture and moxibustion-part 6: acupoint injection].

Some issues on the formulation of Standardized manipulations of acupuncture and moxibustion-part 6: acupoint injection are debated in this paper, including the definition of acupoint injection, characteristics of the standard and application of technical index etc. In the paper, the authors suggest that the issues on the selecting acupoints and drugs should be paid attention during the standard in use. Finally, the authors propose that the technical description of the acupoint injection in the reported literatures needs to be improved in order to better serve for clinical and scientific research.

[Effects of enteral nutrition and parenteral nutrition on gut epithelial tight junction and barrier function in rats after surgical stress].

OBJECTIVE: To compare the effects of enteral nutrition (EN) and parenteral nutrition (PN) on gut mucosal barrier function and intestinal epithelial tight junctions in rats after surgical stress. METHODS: Fifty SD rats with surgical trauma were randomly divided into three groups: total parenteral nutrition (TPN) group and enteral nutrition (EN) group with isocaloric and isonitrogenous nutrition and placebo group. Nutrients were administered via the neck vein and needle jejunostomy for the TPN group. The homogenated tissues of liver, lung, and mesenteric lymph nodes were cultured to determine the bacterial translocations rates. The transmembrane binding proteins (occludin) were measured with immunohistochemistry. The ultrastructure and morphology of intestinal epithelial tight junctions were observed by electron microscope. The feces in cecum were cultured for anaerobic bacterial growth analyses. RESULTS: The EN group had more lactobacteria and bifydobacteria than the TPN group, but not statistical significant. The EN group had greater expression of occludin in the intestines than the TPN group. Furthermore, the intestinal epithelial tight junction and microvilli of the EN group were more intact compared with those of the TPN group. The bacterial translocations rates of liver, lung and mesenteric lymph nodes were significantly lower in the EN group than in the TPN group. CONCLUSION: Neither EN nor standard PN maintains intestinal membrane barriers. But EN increases the expression of transmember binding proteins, maintains the gut epithelial tight junction, improves intestinal mucosal barrier and reduces gut bacterial translocation.

A brief on pain and its patterns in Chinese medicine.

Pain is the most common symptom in clinics. Therefore, the quality, time course and location of pain are important for the diagnosis and treatment. However, pain is very difficult to define, and it is one of the conditions poorly understood by the medical sciences. In Western medicine pain is often described in terms of a penetrating or tissue-destructive process and/or emotional reactions, but in Chinese medicine the pathogenesis and description of pain are different. It is described based on the theories and pathogenesis of Chinese medicine, and the patients' feeling according to their languages and cultures. Thus, when taking the history and conducting physical examination in patients in China and treating them according to Chinese medicine, we have to know how these patients express and describe their pain when they go to clinics. This article will explain some terms and words in the Chinese language and their corresponding terms in the English language.

[Influence of ecoimmunonutrition supplement on intestinal mucosa morphology and gut barrier function in rats after operative stress].

OBJECTIVE: To investigate the effects of ecoimmunonutrition supplement on intestinal microecology, epithelial tight junctions, and barrier function in rats with surgical stress. METHODS: Seventy SD rats after surgical trauma were randomly divided into four groups:(1) placebo group,(2)total parenteral nutrition(TPN) group,(3)enteral nutrition(EN) group and (4)ecoimmunonutrition (EEN)group respectively. Rats received isocaloric and isonitrogenous nutrition. Nutrients were administered via the neck vein and the needle jejunostomy for five days. The homogenated tissues of liver, lung, and mesenteric lymph nodes were cultured to determine the bacterial translocation rate. The transmembrane binding proteins(occludin) was measured by immunohistochemistry. The ultrastructure and morphology of intestinal epithelial tight junctions in the intestine were observed by electron microscope. The feces in cecum was cultured for anaerobic bacterial growth and analysed. RESULTS: The amounts of lactobacteria and bifidobacteria in EEN group were significantly higher than those in TPN group(P<0.05). The expression levels of occludin in the intestine was significantly higher in EEN group than that in TPN and EN group. Furthermore, the intestinal epithelial tight junction and microvilli of EEN group were more intact compared with those of TPN group. The bacterial translocation rates of liver, lung and mesenteric lymph nodes were significantly lower in EEN and EN group than those in TPN group(P<0.05). CONCLUSION: Application of ecoimmunonutrition can protect intestinal mucosal barrier in rats with operative stress, increase the expression of occludin, maintain the gut epithelial tight junction, and eliminate gut bacterial translocation.

The growth inhibitory effect of mesenchymal stem cells on tumor cells in vitro and in vivo.

Mesenchymal stem cells (MSCs) play an important role in the development and growth of tumor cells. The purpose of this study is to confirm the effect of MSCs on tumor cell growth in vitro and in vivo and to elucidate the mechanism. MSCs were isolated from mouse bone marrow and cocultured with murine hepatoma H22, lymphoma (YAC-1 and EL-4) and rat insulinoma INS-1 cell lines. The growth inhibitory effect of MSCs on tumor cells was tested through MTT and 3H-TdR incorporation assay. The apoptosis induction effect of MSCs on tumor cells was assessed with flow cytometry (FCM) and RT-PCR assay. MSCs were inoculated into BALB/c mice alone or coinoculated with ascitogenous hepatoma cells intraperitonealy, respectively. The tumor growth inhibition of MSCs was investigaed through the incidence and volume of ascites formation, and the immunosuppression effect was studied with splenocyte response to ConA stimulation test and T cell subsets analysis (FCM). The results showed that MSCs exhibited a number-dependent growth inhibitory effect on murine tumor cell lines in vitro and inhibited the growth of ascitogenous hepatoma cells in vivo without host immunosuppression. MSCs could upregulate tumor cells mRNA expression of cell cycle negative regulator p21 and apoptosis associated protease caspase 3. The findings of this experimental study demonstrated that MSCs had potential inhibitory effects on tumor cell growth in vitro and in vivo without host immunosuppression, by inducing apoptotic cell death and G(0)/G(1) phase arrest of cancer cells.

[Effects of ErbB signal and protein kinase B on monkey cardiocyte apoptosis induced by rapid pacing].

This study is to investigate the roles of neuregulin receptor ErbB2 and protein kinase B (PKB) in pacing-induced heart failure of rhesus monkey. Rapid pacing was used to induce heart failure in rhesus monkey. Aorta intubatton was used to perform hemodynamic measurements, 17 days after pacing. N-Terminal pro-brain natriuretic peptide (BNP), one of the most important molecular marker of heart failure, was also measured by the method of electrochemical luminescence immunoassay. The mRNA expressions of ErbB2, PKB and Bcl-xl were detected in the left ventricular free walls by RT-PCR method. The expressions of phospho-PKB and Bcl-xl on protein level were also detected by Western blotting. The contractibility of cardiac muscle decreased significantly, which consisted with the increase of BNP. Compared with control group, the mRNA expressions of ErbB2, PKB and Bcl-xl were depressed, and similar results were also found in the protein expression analysis of phospho-PKB and Bcl-xl. The expressions of ErbB2, PKB and Bcl-xl were down-regulated during heart failure in rhesus monkey which suggested the important roles of ErbB2 receptor and PKB in the mechanism of heart failure.

[Systematic assessment of acupuncture for treatment of herpes zoster in domestic clinical studies].

OBJECTIVE: To assess the effectiveness of acupuncture for treatment of herpes zoster. METHODS: According to the requirement of evidence-based medicine, acupuncture, body acupuncture, electroacupuncture, head acupuncture, three edged needle, plum-blossom needle, fire needle, elongated needle, encircling needling, herpes zoster, etc. were selected as subject words to retrieve the relative medical database at home, and clinically randomized controlled trials were used as enrolled criteria, the treatment group were treated with acupuncture or acupuncture plus other therapies, and the control group with medicine, the cured rate and the time of killing pain for herpes zoster were used as assessment indexes. Altogether 43 papers were enrolled. Among them 10 papers were conducted for Meta-analysis by RevMan 4.2.9. RESULTS: The total OR was 4.27 with 95% CI [2.90, 6.29] of the clinically cured rate in the 10 studies, and the total OR was -7.64 with 95% CI [-8.12, -7.15] of the time of killing pain in the 4 studies. The therapeutic effect in the treatment group on herpes zoster was superior to that of the western medicine (P < 0.01). CONCLUSION: Acupuncture therapy for herpes zoster is effective, but more high-quality studies are required to prove this view point.

[Construction of a cDNA library from liver tissue of rhesus monkey, Macaca mulatta].

OBJECTIVE: To screen the target rhesus genes and give some basic genetic evidences to its value as one of the most important animal model in biomedical study, we constructed a cDNA expression library from liver tissue of a healthy rhesus monkey. METHODS: With Trizol reagent, the total RNA was extracted from healthy rhesus liver tissue. By mutant Moloney Murine Leukemia Virus Reverse Transcriptase (MMLV-RT), the first-strand cDNA was synthesized from purified mRNA, and subsequently the second-strand cDNA was generated via E. coli DNA polymerase I . Then, the EcoR I adapter was added to the synthesized double-strand cDNA, which was subsequently digested by Xho I restriction enzyme and fractionated with CHROMA APIN-400 column. The fractionated cDNA fragments to be longer than 0. 5 kb were ligated into lambda ZAP express vector to form the phagemid cDNA recombinants, which were further packaged into the lambda ZAP cDNA library according to the standard protocol with phage lambda Gold packaging extract. In order to get more stable clones with larger quantity, the primary library was amplified through infecting the host strain XL1-Blue MRF'. Then, the library titre, recombinant rate and length of inserted cDNA were measured, respectively. RESULTS: The capacity of the primary stand or unamplified library was 1. 2X 10(6) pfu. The titers of the unamplified library or the amplified library was 1.1 X 10(6) mixture, pfu/mL or 7. 7 X 10(9) pfu/mL respectively, the percentages of recombinants were 99. 3% and 98. 2%, and the average lengths of the inserts were 2.0 kb and 2. 3 kb, respectively. CONCLUSION: An excellent cDNA expression library has been constructed successfully, which would lay solid foundation for transplantation study and pre-clinic evaluation of related drugs.

[On relationship of acupoint-injection with injury of peripheral nerves].

OBJECTIVE: To provide basis for strengthening safety of acupoint-injection and increasing clinical therapeutic effect. METHODS: Analyze and study on the relative articles from the databank of whole articles of Chinese periodicals of CNKI by information retrieval with computer, with acupoint-injection, nerve injury as key words. RESULTS: Most of clinical reports focus on acupoint-injection for treatment of nervous injury induced by trauma and birth injury. The studies indicate that the injuries of the peripheral nerves induced by acupoint-injection can be divided into 3 grades and the injury mechanisms can be divided into 3 classifications. The injuring causes include improper posture of the patients, improper angle and depth of injection and improper medicine selection. CONCLUSION: Acupoint-injection can be applied more widely as soon as the accomplishment of the standardization of operation.

Expressed sequence tags analysis of a liver tissue cDNA library from a highly inbred minipig line.

BACKGROUND: Porcine liver performing efficient physiological functions in the human body is prerequisite for successful liver xenotransplantation. However, the protein differences between pig and human remain largely unexplored. Therefore, we investigated the liver expression profile of a highly inbred minipig line. METHODS: A cDNA library was constructed from liver tissue of an inbred Banna minipig. Two hundred randomly selected clones were sequenced then analysed by BLAST programme. RESULTS: Alignments of the sequences showed 44% encoded previously known porcine genes. Among the 56% unknown genes, sequences of 72 clones had high similarities with known genes of other species and the similarities to human were mostly above 0.80. The other 40 clones showing no similarity to genes in National Centre for Biotechnology Information are newly discovered, expressed sequence tags specific to liver of inbred Banna minipig. Twenty-two of the 200 clones had full length encoding regions, 38 complete 5' terminal sequences and 140 complete 3' terminal sequences. CONCLUSION: These newly discovered expression sequences may be an important resource for research involving physiological characteristics and medical usage of inbred pigs and contribute to matching studies in xenotransplantation.