The standardization of syndrome differentiation based on the diseases of WM has become an important academic guidance,which,however,has induced lots of problems.The logical contradiction was analyzed between the results of standardization of syndrome differentiation based on the diseases of TCM and WM,between the results of two large-scale standardizations of syndrome differentiation based on WM and between the results of syndrome differentiation of related diseases of WM.In this paper,the author studied on the origin of symptoms and signs of syndrome differentiation of the diseases of WM and the effect of standardization of syndrome differentiation of the diseases of WM on the nature and location of disease in TCM,and pointed out some important unresolved academic problems.

Objective: To study the effects of zinc-deficiency(ZD)and zinc-supplement(ZS)on learning and memory in aging mice. Methods: Seventy 3-month-old male mice were divided randomly into 5 groups:normal young , aging model , aging+ZD, aging+pair-fed , aging+ZS . The aging model was established with D-galactose (100 mg/kg bw) by nape subcutaneous injection, while the young control group was injected with normal saline in the same way, which lasted 30 days. Aging+ZD and aging+ZS group were fed with ZD feed(zinc 1.61 mg/kg), others with normal feed(zinc 50 mg/kg). ZS feed(zinc 100 mg/kg)was given to aging+ZS for last 2 weeks. During the last week, the behavior tests were carried on. Finally the mice were all killed and samples were collected immediately for detection of indices . Results:Compared with the aging model group, ZD made serum zinc decrease significantly, the locomotor activity reduced and the ability of learning and memory weakened. Comet assay also showed that ZD induced DNA damage of cerebrum cells more serious in aging mice than the control. The ratio of tail length/comet cell length in Group ZD also increased significantly. However, moderate ZS could turn those indices better significantly. Conclusion: Zinc can ameliorate the learning and memory function of aging significantly. By reducing the function of DNA repair, ZD can act on learning and memory, while moderate ZS seems to be beneficial.

Objective To observe the transport of hepatitis B virus (HBV)-infected peripheral blood mononuclear cells (PBMC)through placental barrier set up by choriocarcinoma trophoblast cells (Bewo cells),and to explore the biological role of PBMC as a carrier for HBV transport.Methods Bewo cells and PBMC were cultured and their proliferation and activity were detected by cell counting kit (CCK)-8.One hundred μL serum containing 5 ×10 6 copy/mL HBV DNA was used to infect PBMC,and cells infected with HBV were labeled by fluorescent dye carboxyfluorescein diacetate succinimidyl ester (CFSE).A co-culture model of Bewo cells and HBV-infected PBMC was set up by transwell chamber. The migration of HBV-infected PBMC was detected by flow cytometry.Realtime fluorescence quantitative polymerase chain reaction method was used to detect HBV DNA contents of PBMC under transwell chamber.Results PBMC and Bewo cells proliferated at around 24 h and entered into growth stagnation at around 120 h.The contents of PBMC labeled by green fluorescent at 0,12,24 and 48 h during co-culture under chamber were (0.445 ±0.021)%,(21 .180 ± 4.653 )%,(34.830 ± 7.156 )% and (64.185 ± 3.161)%,respectively.The amount of PBMC marked green fluorescence increased over prolonged incubation time (F =68.983,P =0.001 ).PBMC HBV DNA contents at 24 and 48 h of co-culture under chamber were (1.925±0.431)×103 copy/mL and (2.565 ±0.361)×103 copy/mL,respectively,indicating that PBMC under chamber were infected with HBV.Conclusions PBMC may be a target for HBV infection in extrahepatic tissues.Placental trophoblastic barrier built by transwell chambers may provide new ideas to investigate HBV transmission across the placenta in vitro .

BACKGROUND:On-pump coronary artery bypass grafting(CABG)is replaced by off-pump coronary artery bypass grafting step by step.Effects of the two operations on liver and renal function deserve further studies.OBJECTIVE:To analyze the postoperative changes in liver and renal function between off-pump and on-pump CABG.DESIGN.TIME AND SETTING:Controlled study.The experiment was conducted at the Department of Cardiovascular Surgery.General Hospital of Chinese PLA.Beijing between June and November 2005.PARTICIPANTS:Fifty patients undergoing elective CABG at General hospital of PLA from June to November 2005 were enrolled in the study.All patients had normal liver and renal function before surgery.None of them developed severe acute liver and renal function.METHODS:All patients were allocated to either off-pump coronary bypass(OPCAB)(n=30)or on-pump conventional CABG (CCABG)(n=20)group according to patient's intention and condition.No significant difference was detected in age,gender,body mass index,preoperative ejection fraction,preoperative liver and renal function and operation risk factors between both groups(P>0.05).MAIN OUTCOME MEASURES:Alanine aminotransferase(ALT),aspartate aminotransferase(AST),blood urea nitrogen (BUN)and creatinine(Cr)were respectively measured in patients of both groups before surgery,1 day,1 week and 2 weeks after surgery.RESULTS:A total of 50 patients were involved in the final analysis.Changes in liver function:Serum ALT and AST Ievels in the OPCAB group were significantly lower than those in the CCABG group at 1 day after surgery(P<0.05).The level of two parameters recovered to the baseline at 2 week after surgery in both group.Changes in renal function:Serum BUN and Cr levels in the OPCAB group were significantly lower compared to the CCABG group at the first postoperative day(P<0.05).BUN and Cr levels recovered to the preoperative levels at 2 weeks after surgery in bOth group.CONCLUSl0N:CABG has an adverse effect on liver and renal function.which can recover in the early postoperative period.OPCAB has the less adverse effect on liver and renal function and offers a better liver and renal protection compared to CCABG.

Objective To observe the effects of zinc-deficiency(ZD)on antioxidation system and repair of DNA damage and of hepatic cells by establishing an aging mice model induced with D-galactose. Methods Seventy 3-month-old male mice were divided randomly into 5 groups: normal young group, aging model group, aging+ZD group, aging+pair-fed group, aging+ZS group. The aging model were established with D-galactose (100mg/kg bw) by nap subcutaneous injection, while the young control group were injected with normal saline in the same way which lasted 30 days. Aging+ZD group and aging+ZS group were fed with ZD food(zinc 1.61?g/kg), others with normal food(zinc 50?g/kg). ZS food (zinc 100?g/kg)were given to aging+ZS group for the last 2 weeks in the course. On the 30 th day, all mice were killed by removing eyeballs and samples were collected immediately for later indexes detection. Results Compared with the aging control group, zinc contents and SOD activity of serum and liver decreased significantly, while MDA in the serum and liver and lipofuscin of liver increased much more in ZD mice. Comet assay also showed that the DNA damage of hepatic cells were more serious in aging ZD mice than in the control mice. The ratio of tail length/comet cell length in group ZD also increased significantly. However, moderate zinc supplement significantly improved all those indexes. Conclusions Zinc can affect the speed and degree of aging significantly. Zinc deficiency accelerates the aging progress; while moderate zinc supplement seems to be beneficial for retarding aging progress.

Objective To investigate the effects of inducible hypothermia on expressions of peroxisome proliferator activated receptor gamma coactivator-1 (PGC-1) and small heterodimer partner (SHP) in rat model of hemorrhagic shock.Methods SD rats were randomly (random number) divided into three groups:control (C),normothermia (N) and hypothermia (H).Exsanguination was carried out in rats by continuously drawing out venous blood (25 mL/kg) over 15 minutes to establish hemorrhagic shock model.Then,rectal temperatures of rats were maintained by body surface cooling to 32℃ in H group and by body surface warming to 38℃ in N group,respectively.After a shock period of 60 minutes,rats received the infusion of whole blood of their own and lactated Ringer's solution (1 ∶ 2) treatment for 60 min.Rats were warmed to 38℃ by body surface warming and monitored for 3 h after resuscitation.Hematocrit (Hct),base excess (BE),lactate (Lac),and glucose (Glu) were recorded before modeling and after different lengths of hemorrhagic shock period (HSP).The expressions of PGC-1 mRNA and SHP mRNA and the levels of their protein in liver were tested by real-time reverse transcription polymerase chain reaction (qRT-PCR) and western blotting,respectively.Results The H group had lower lactate levels and higher BElevels than the N group [(6.3±2.1) vs.(10.4±1.5) and (-4.7±2.5) vs.(-9.0±3.2)] (P＜ 0.05).At 72 hours after modeling,there were four survivors in the N group and seven survivors in the H group (P ＜ 0.05,Log Rank).The expressions of PGC-1 mRNA and SHP mRNA increased in N group.Hypothermia resuscitation down-regulated PGC-1 mRNA expression,meanwhile,increased expression of SHP mRNA.Both Hypothermia and Normothermia resuscitation increased SHP protein levels,but decreased PGC-1 protein levels.Conclusions Inducible hypothermia ameliorated acidosis and energy metabolism imbalance through adaptive regulation in PGC-1 and SHP.

BACKGROUND:Many studies have demonstrated that FirebirdTM and Taxus Express2TM can effectively reduce intrastent restenosis.However,there are few data about long-term efficacy of two stents,and reports about middle and long-term follow up are rare.OBJECTIVE:To observe the safety and biocompatibility of FirebirdTM and Taxus Express2TM in percutaneous coronary intervention(PCI) for coronary artery disease.DESIGN,TIME AND SETTING:Non-randomized concurrent control clinical observation was performed at Department of Cardiology,Hongqi Hospital of Mudanjiang Medical College from April 2005 to April 2008.PARTICIPANTS:233 patients(268 lesions) undergoing PCI were divided into FirebirdTM(n =82),Taxus Express2TM group(n =80) and bare metal stent group(n =71).METHODS:Coronary arteriography was performed through femoral artery or radial artery.Vascular inner diameter was determined using quantitative computer analysis.The patients underwent FirebirdTM,Taxus Express2TM and bare metal stenting,respectively.The patients were reexamined and followed-up using telephone every 2-4 weeks after discharging and examined using coronary arteriongraphy after 9-12 months.The follow-up lasted for 24 months.MAIN OUTCOME MEASURES:Characteristics of arteriongraphy and stenting condition of all patients;biocompatibility of stent to host;major adverse cardiac events during hospitalization and follow-up,including death,angina pectoris attacks or heart failure;coronary artery diameter decreased ≥ 50% was regarded as restenosis.RESULTS:101 stents were implanted in Firebird group,98 in Taxus Express2 group and 85 in bare metal stent group.There was no stent defluxion,dislocation,or breakage.No noticeable platelet decrease,hemolysis or white blood cell increase was found.There were no significant differences among three groups in terms of Characteristics of arteriongraphy and stenting condition.The incidence of major adverse cardiac events and intravascular restenosis in Firebird and Taxus Express2 groups was fewer than bare metal stent group(P 0.05).CONCLUSION:No specific biocompatibility responses in treatment of coronary artery diseases using FirebirdTM and Taxus Express2TM.The two drug-eluting stents are superior over bare metal stent in reducing restenosis.The safety and efficacy of two drug-eluting stents are similar.

Objective To analyze the feasibility of the recombinant cholera toxin B subunit (rCTB) as a carrier protein candidate for the preparing of polysaccharide-protein conjugate, and to discuss the immune effects of tetanus toxoid (TT) as the carrier protein in mucosal delivery vaccine. Methods The refolded pentrumer protein, rCTB was obtained by genetic engineering methods. Then conjugated the refold-ed protein with group A meningococcal polysaccharide (GAMP) using the chemical method(ADH) ,the pol-ysaccharide-protein conjugates(GAMP-rCTB) were prepared. BALB/c mice were immunized either intraper-itoneally ( i. p. ) or intranasally ( i. n. ) with GAMP-rCTB. Moreover, GAMP-TT vaccine that TT as carrier proteins was i.n. immunized to the mice. The evaluation of immunology is performed. Results The conju-gates of polysaccharide-potein with the rCTB and TT as protein carrier both are able to elicit high level of GAMP specific IgG antibody in serum after i.n. immunization, and the conjugates can also elicit specific IgA antibody in lung lavage and intestinal mucosa. Conclusion rCTB and TT can both as the protein carri-er for polysaccharide-protein conjugate as mucosal vaccine. The route of intranasal may be more ways for im-mune function than i.p. immunization when rCTB is used as the carrier of the polysaccharide-protein conju-gates.

Objective To establish a LC-MS/MS method for the determination of MMA in serum,and provide a assay for the diagnosis and screening of methylmalonic academia in the clinic. Methods MMA was extracted from 205 serum samples from healthy controls and 146 serum samples from patients with liquidliquid extraction method with MTBE as the extraction solvent. The supernatant was transferred to a tube and dried with nitrogen gas. Then the residual was derived with HCI-BuOH mixed agent to give a product, which was analyzed directly by LC-MS-MS system with a gradient elution, selective reaction monitor, a Discovery C18 column (50 mm × 2. 1 mm ,5 μm) as the isolation column and a mobile phase consisting of methanol and water (0. 1% formic acid, V/V), respectively. The concentration of MMA was detected with the isotope internal standard method. The stand curve was employed with a series of calibrators. The recovery was estimated with the 3 serum samples with the concentrations of 2, 25, 80 μg/L respectively. The accuracy,precision and stability were also tested with quality control samples. Moreover, the range of concentrations in healthy people were detected to investigate the influence of hemolysis on the detection results. Thirteen samples were randomly tested according to the digital chart. The testing results were compared with the results provided by Medical Diagnositic institution (MDI) in Germany. The paired t-test was applied to statistical analysis. Results The linear range of this method was 2-100 μg/L, and the correlation coefficient (R2 ) was more than 0. 995. The retention time of MMA derivative was 10. 5 min. Succinic acid and MMA were not found to interfere with each other. The within-run RSD was less than 6. 4%, and the between-run RSD was less than 5.0%. The recovery rates were from 96. 42% to 103. 33%. The limit of quantification was 1 μg/L.The accuracies of the method were form 94. 2% to 108. 2%. The samples were stable for 6 h at room temperature and stable for 70 d even keep at - 20 ℃. The samples were stable after 10 freeze-thaw cycles. The derivatives of MMA were found to be stable at least for 5 d at 4 ℃. The medians of the hemolysis group and the normal group were 102.53 (13.84-302.33) μg/L and 39.52 (11.94-203.08) μg/L,respectively. There was significant difference between the 2 groups ( T = 8, P ＜ 0. 05 ). The medians of comparison test in our laboratory and the MDI were 32. 82(24. 50-100. 42) μg/L and 32. 20(26. 65-93. 30)μg/L There was no significant difference between the 2 groups( T=7 ,P ＞0. 05 ). The mean value (-x± s)of 158 healthy adults( 18-58 years old) and 47 healthy teenages( 1-17 years old)were ( 18.46 ± 10.49 )μg/L and (22. 38 ± 11.45) μg/L, respectively. Conclusions A LC-MS/MS method for analysis of MMA in serum is established successfully. The quantitative method is simple and accurate with good sensitivity,specificity and repeatability. The method can be applied for diagnosis, screening and monitoring of methylmalonic acidemia.

Aim To determine the function of nuclear factor-κB ( NF-κB ) in immunological liver injury of rat model and its effect on CYP2E1 expression, content and metabolic activity. Methods The immunological liver injury rat model was prepared by injection of Ba-cillus Calmette Guérin ( BCG,125 mg · kg-1 ) for 14 days. The hepatic tissue injury was revealed by hema-toxylin and eosin ( HE ) method and serum concentra-tion of alanine aminotransferase( ALT) , aspartate ami-notransferase ( AST ) respectively. CYP450 total con-tent in hepatic homogenate was determined by spectro-photography. The expression of CYP2E1 protein was detected by Western blot analysis. The enzyme kinetics of CYP2 E1 probe drug chlorzoxazone was evaluated by high-performance liquid chromatography ( HPLC ) as-say. Results The results showed that BCG-pretreat-ment ( 125 mg · kg-1 ) significantly increased the weight of liver and spleen, serum levels of ALT and AST(P<0. 01) , and decreased CYP2E1 expression, content and metabolic activity ( P <0. 05 ) . Adminis-tration of ammonium pyrrolidine dithiocarbamate (PDTC) (50, 100, 200 mg·kg-1) reversed the a-bove hepatic injury stimulated by BCG in vivo. Moreo-ver, PDTC dose-dependently inhibited the down regu-lation of CYP2 E1 ( P<0. 05 ) . Conclusion Passiva-tion of NF-κB can inhibit the down regulation of CYP2 E1 in liver tissue of immunological liver injury rats;NF-κB may be involved in CYP2 E1 down-regula-tion.