AIM:To investigate the ability of a metal complex ammonium tetrathiomolybdate (ATTM) to re-lease H2 S and its cytoprotective effect on an oxidative injury model .METHODS:Released H2 S was absorbed in a reaction flask from ATTM dissolved in the cell medium .Staining with dichlorodihydrofluorescein diacetate or rhodamine 123 fol-lowed by photofluorography was conducted for the observation of reactive oxygen species ( ROS) and mitochondrial mem-brane potential (ΔΨm) levels, respectively.Cell viability and release of lactate dehydrogenase (LDH) from the cells were measured with commercial kits.RESULTS:Similar to another H2S donor GYY4137, ATTM had an ability to release H2S in the cell medium in a dose-dependent manner .Treatment of human skin HaCaT cells with ATTM at concentrations of 25~400 μmol/L didn’ t significantly alter cell viability .Exposure of the cells to ultraviolet rays or a ROS donor H 2 O2 in-creased the intracellular ROS levels .Treatment with 400 μmol/L H2 O2 significantly reduced the viability of HaCaT cells (P<0.01).However, before the treatment with H2O2, pretreatment with ATTM at 100 and 200 μmol/L markedly pre-vented the H2O2-induced cell injury (P<0.01).In addition, the treatment with H2O2 triggeredΔΨm loss (P<0.01) and LDH release from the cells (P<0.01).Prior to suffering from H2O2 injury, the preconditioning with 200 μmol/L ATTM significantly improved ΔΨm levels ( P<0.05 ) and attenuated LDH release from the cells ( P<0.01 ) .CONCLUSION:ATTM is capable of releasing H 2 S and protecting human skin cells against oxidative injury .

Objective:To explore the expression of TNF a and IL-10 in different layers of laryngeal tissues after laryngeal transplantation and its relationship with acute rejection.Methods:Laryngeal heterotopic transplantation was performed in Wistar and SD rats(Wistar→SD rats).The SD rats were divided into 4 groups:GroupⅠ:Sham control(receive no transplantation): GroupⅡ(receive transplantation,without cyelosporine A treatment);GroupⅢ(receive transplantation.with 5 mg?kg~(-1)?d cyelosporine A treatment):and GroupⅣ(receive transplantation.with 10 mg?kg~(-1)?d~(-1)cyelosporine A treatment).The transplanted larynx was harvested on 3,7 and 11 days after transplantation for pathological examination.The expression of TNF-?and IL-10 in different layers of grafts was detected immunohistochemically.Results:Pathological observation showed mild,moderate and severe acute rejection in GroupⅡandⅢon 3.7 and 11 days after transplantation,respectively:there was no obvious rejection in GroupⅣ.Immunohistochemical staining showed expression of TNF-?and IL-10 in GroupⅡ.Ⅲ.andⅣ,not in GroupⅠ.The ratios of the positive areas of TNF-?and IL-10 in the mucosal and submucosal layers were significantly higher than those in the muscle and cartilage layers of laryngeal tissues(P

To explore a new method for identification of Mongolian patent medicine (MPM) by PCR amplification of specific alleles. Eight kinds of MPM were used to study the identification of "Digeda" raw materials. The total DNA of Lomatogonium rotatum and Corydalis bungeana samples were extracted through modified CTAB method, psbA-trnH sequence was amplified by PCR and sequenced directionally. Specific primer was designed. The DNA of 8 kinds of MPM also was extracted and purified by the commercial DNA purification kits. The rbcL and two pair of specific primers sequences were amplified. The specific amplified products were sequenced in forward directions. All specific sequences were aligned and were analyzed. The results indicated that L rotatum can be identified by specific primers from Digeda-4 Tang, Digeda-8 San, Digeda-4 San, and C. bungeana medicinal materials can be identified by specific primers from Li Dan Ba Wei San, Yi He Ha Ri-12 and A Ga Ri-35. PCR amplification of specific alleles can stably and accurately distinguish raw medicinal materials in MPM.
Alleles ; DNA Primers ; genetics ; DNA, Plant ; genetics ; Medicine, Mongolian Traditional ; Molecular Sequence Data ; Plants, Medicinal ; classification ; genetics ; Polymerase Chain Reaction ; methods

OBJECTIVETo explore the correlation between the recurrence of cerebral infarction and aspirin resistance (AR)/Chinese medical (CM) constitutions.

METHODSTotally 413 cerebral infarction patients took Aspirin Enteric-coated Tablet (100 mg per day) while receiving routine therapy, 5 days at least in a week. They were followed-up for 12 months. Aspirin sensitivity (AS) was determined using turbidimetry. CM constitutions among patients with different AS were compared. Ratios of AR patients and AS patients of different CM constitutions in cerebral infarction recurrent patients were compared. Platelet membrane glycoproteins (GP) II b HPA-3 gene polymorphism was detected by polymerase chain reaction (PCR) method. Correlation between recurrence of cerebral infarction and AR, bb genotypes, CM constitutions times AS were analyzed by Logistic regression.

RESULTSTotally 11 patients dropped out, 101 (25.12%)with recurrent cerebral infarction and 301 (74.88%) without recurrent cerebral infarction. There were 152 (37.81%) AR patients and 250 (62.19%) AS patients. AR accounted for 26.6% (80/ 301) and AS accounted for 73.4% (221/301) in non-recurrent cerebral infarction patients. AR accounted for 71.3% (72/101) and AS accounted for 28.7% (29/101) in recurrent cerebral infarction patients. There was statistical difference in AR and AS ratios (χ2 = 64.287, P = 0.000). The proportion of yin deficiency constitution (YDC) was the largest [28.3% (43/152)] in AR patients. The proportion of blood stasis constitution (BSC) was the largest [23.6% (59/250)] in AS patients. There was statistical difference in CM constitutions between AR patients and AS patients (χ2 = 21.574, P < 0.01). The former 4 recurrent rates occurred in AR patients of YDC, BSC, damp-phlegm constitution (DPC), qi deficiency constitution (QDC). YDC occupied the first place [22.4% (34/152)]. The former 4 recurrent rates occurred in AS patients of BSC, QDC, DPC, damp-heat constitution (DHC). BSC occupied the first place [3.2% (2/250)]. Compared with non-recurrent cerebral infarction patients and AS patients, bb gene occurred most often, but aa gene and ab gene occurred obviously lesser in non-recurrent cerebral infarction patients and AR patients (χ2 = 20.171, χ2 = 55.139, P < 0.01). AR and bb gene were positively correlated with recurrent cerebral infarction (OR = 18.423, P = 0.000; OR = 1.304, P = 0.028). Body constitutions interacted with AS (OR = 0.707, P = 0.000).

CONCLUSIONSRecurrent cerebral infarction was closely related to AR and constitutional types. The recurrence rate was higher in AR patients of YDC. GP I b HPA-3 bb genotype might be a risk factor for AR and recurrent cerebral infarction.

Aspirin ; therapeutic use ; Body Constitution ; Cerebral Infarction ; Drug Resistance ; Humans ; Medicine, Chinese Traditional ; Neoplasms ; Recurrence ; Yin Deficiency

To silence the expression of K-RASAsn12 in human pancreatic cancer cell line by vector-based RNAi(RNA interference) technique,two single-strand DNA sequences encoding mutant-specific shRNA (short haipin RNA) for K-RASAsn12 were synthesized and then inserted into pSilenCircle. The recombinant plasmid was called pSC-K-RASAsn12. According to the same method, pSC-GFP encoding shRNA for GFP was gained. Both recombinant plasmids were transfected into human pacreatic cancer cell line AsPC-1 and BxPC-3. The expression level of K-RASAsn12 was detected by semi-quantitative RT-PCR and Western blot. The result indicated that the recombinant plasmid edcoding mutant-specific shRNA for K-RASAsn12 can inhibit significantly the expression of K-RASAsn12 without affection of wild-type K-RAS(K-RASWT)in Human Pancreatic Cancer Cell Line.

Objective To detect the Yersinia pestis plasmid and molecular weight along the Qinghai-Tibet Railway.Methods Yersinia pestis plasmids molecular weight detected and analyzed using alkaline lysis,phenol-chloroform extraction of Yersinia pestis plasmid by agarose gel electrophoresis.Results The 18 Yersinia pestis strains of Qinghai-Tibet Railway contained 6×106,45×106,52×106,65×106,92×106plasmid,varing in the range of the 52×106-92×106.Conclusions The Yersinia pestis of Qinghai-Tibet Railway has a standardplasmid graphics,with the biggest Yersinia pestis plasmid changing in a certain regular degree,which providessignificance in the study of plague natural foci of the spatial structure and the genetic.characteristics of Yersiniapestis.

ABO Blood-Group System ; Erythroblastosis, Fetal ; epidemiology ; Humans ; Infant, Newborn

OBJECTIVETo explore the mechanism of the eye-acupuncture for treatment of acute cerebral ischemia-reperfusion injury.

METHODSThirty-two healthy SD rats were randomly divided into a normal group, a sham operation group, a model group and an eye-acupuncture group, 8 rats in each group. The rat model of cerebral ischemia-reperfusion was established with thread occlusion method in the model group and the eye-acupuncture group. The eye-acupuncture group was treated by eye-acupuncture at "liver region", "upper energizer area", "lower energizer area" and "kidney region" for 20 min immediately after reperfusion and at 30 min before sampling. No treatment was done in the normal group and the sham operation group, and no thread occlusion was performed in the sham operation group. The Neurologic impairment was scored and the methods of immunohistochemistry staining, western-blotting and real-time fluorescent quantitation polymerase chain reaction (RQ-PCR) were taken to detect the expression of the aquaporin protein 4 (AQP4) and its mRNA in cerebral cortex after reperfusion for 3 hours.

RESULTSThe neurologic impairment score of 1.50 +/- 0.54 in the eye-acupuncture group was significant lower than 2.63 +/- 0.92 in the model group (P < 0.01). The expression of the AQP4 protein by immunohistochemistry and western-blot respectively were 116.33 +/- 10.24 and 0.53 +/- 0.04 in the normal group, 118.97 +/- 12.72 and 0.55 +/- 0.07 in the sham operation group, and 129.30 +/- 18.36 and 0.67 +/- 0.08 in the eye-acupuncture group, with statistical significance compared to 150.88 +/- 15.82 and 0.94 +/- 0.04 in the model group (all P < 0.01), and there were significant differences between the eye-acupuncture group and the normal group (both P < 0.01). The tendency in the expression of AQP4 protein and its mRNA in all the group were almost the same.

CONCLUSIONThe eye-acupuncture therapy can relieve the cerebral ischemia-reperfusion injury and the protective mechanism is related to the downregulation of the cerebral AQP4 expression.

Acupuncture Therapy ; Animals ; Aquaporin 4 ; genetics ; metabolism ; Brain ; metabolism ; Brain Ischemia ; complications ; genetics ; metabolism ; surgery ; Disease Models, Animal ; Eye ; anatomy & histology ; Female ; Gene Expression ; Humans ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; etiology ; genetics ; metabolism ; therapy

Acute Disease ; Animals ; Ginkgo biloba ; Glutathione ; analysis ; Lung ; drug effects ; metabolism ; pathology ; Malondialdehyde ; analysis ; Paraquat ; toxicity ; Phytotherapy ; Plant Extracts ; therapeutic use ; Poisoning ; drug therapy ; Rats ; Superoxide Dismutase ; analysis ; Treatment Outcome

The effects of femoral nerve electrostimulation (FNES) on ischemia-reperfused myocardium were examined in the urethane- anesthetized rats to determine whether FNES may provide cardioprotection and to observe the possible mechanism. The area at risk (AR) and infarct area (IA) were determined using Evans blue and nitro-blue tetrazolium staining, respectively. Infarct size (IS) was defined as 100xIA/AR (%). The results are as follows: (1) During 30 min myocardial ischemia and subsequent 120 min reperfusion, the myocardial infarct size occupied (54.96+/-0.82)% of the area at risk. (2) FNES of high frequency (10 V, 100 Hz, 1 ms) significantly reduced myocardial infarct size to (36.94+/-1.34)% (P<0.01), indicating the cardioprotective effect FNES of high frequency on myocardial ischemia-reperfusion, while FNES of low frequency (10 V, 10 Hz, 1 ms) had no effect on myocardial infarct size. (3) Pretreatment with either naloxone (5 mg /kg, i.v), a nonselective opioid receptor antagonist, or glibenclamide (5 mg /kg, i.v), a K(ATP) channel antagonist, completely abolished the cardioprotection of FNES (100 Hz) from myocardial ischemia-reperfusion. It is suggested that FNES of high frequency can protect myocardium from ischemia-reperfusion injury. The possible mechanism is that FNES of high frequency may induce the release of opioids from the central nervous system, and the activation of opioid receptors in the heart results in an opening of myocardial K(ATP) channels which can protect myocardium.
Animals ; Electric Stimulation ; methods ; Femoral Nerve ; physiopathology ; Glyburide ; pharmacology ; Male ; Myocardial Infarction ; pathology ; Myocardial Reperfusion Injury ; pathology ; prevention & control ; Naloxone ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, Opioid ; metabolism