The chelating reaction between glutathione peptides and divalent cadmium ion was used as a typical model for investigating the coordination chemistry of SH-containing peptides and heavy metal ions, which was essential to understand the mechanism of intracellular cadmium detoxification.Here, the mutant (CM113R)7 αHL protein nanopore equipped with a first synthesized per-6-quaternary ammonium-β-cyclodextrin (p-QABCD) was used as nanoreactor and detector to investigate the single-molecule reaction of GSH molecules and Cd2+ ions.Different reaction pathways, intermediates, and products could be recognized.Cd2+-GSH reaction was highly dependent on the solution pH.The Cd(GSH)2 was formed at pH 7.4, while the Cd(GSH)2 and Cd2(GSH)2 were formed at pH 9.0.Cd2(GSH)2 was formed by two possible pathways: (1) A Cd2+ ion primarily coordinated with the thiol group of two GSH molecules to form Cd(GSH)2, and then the second Cd2+ ion quickly incorporated with the deprotonated amino group of Cd(GSH)2 to form Cd2(GSH)2;(2) Two Cd2+ ions separately coordinated with the thiol and deprotonated amino group of one GSH molecule to form Cd2(GSH)1, and the second GSH molecule quickly bounded to Cd2+ ions to form Cd2(GSH)2.This work studied the chelating reaction between metal ions and bioactive glutathione at single-molecule level without labeling and chemical modification, and would be important to further understand intracellular mechanisms of detoxification of heavy metals and greatly expand the research field of nanopore single-molecule technique.

Objective To investigate the effects of transplantated tumor capillary permeability under microbubble enhanced diagnostic ultrasound exposure.Methods Fourty eight rats were randomly divided into four groups after Walker 256 carcinomas were subcutaneousely implanted,named the control group (A),microbubbles group (B),ultrasound group (C),microbubbles and ultrasound group (D).Using Evans blue(EB) as the indicator for tumor capillary permeability,the microbubbles were injected through the vein with diagnostic ultrasound frequency as 1.75 MHz,mechanical index as 1.4.The spillover of EB was estimated under confocal laser microscope,meanwhile the contents of EB in the tumor tissues were calculated according to the standard curve and spectrophotometry.Results EB spillover was observed around the capillaries in the group D,while EB spillover was only observable on capillary in group A,B and C.The EB content was also significantly higher than those in group A,B and C.Conclusions Exposure to both diagnostic ultrasound and microbubbles results in increased capillary permeability of rat tumor tissues,which might become a potential therapeutic method on clinical chemotherapy.

Objective To investigate the feasibility of transfecting gene into HepG2 by therapeutic ultrasound combined with microbubble,and to explore the optimal ultrasonic parameters for high transfection efficiency.Methods HepG2 cells were cross-interval planted in 24-well plates.EGFP plasmid DNA and microbubbles were added to the cultured HepG2 and three parameters of the therapeutic ultrasound were optimized.Firstly,set ultrasonic intensity gradient (group A,from A1 to A5),which were 0.4 W/cm,0.8 W/cm2,1.2 W/cm2,1.6 W/cm2 and 2.0 W/cm2 respectively.Secondly,set the duty cycle gradient group (group B,from B1 to B3),which were 10％,20％ and 50％ respectively.Lastly,set the exposure time group(group C,from C1 to C3),which were 30 s,90 s and 180 s.After 24 hours,the expression of GFP was observed by fluorescence microscopy,the transfection rates was detected by flow cytometry,the cell death were assessed by trypan blue exclusion.Results The plasmid transfection efficiency was varied under different ultrasonic parameters.Within a certain range,the increasing of parameters can improve the transfection efficiency,but when the parameters were too large (eg.the intensity＞1.6 W/cm2 or 50％ duty cycle),the actual transfection efficiency decreased due to the increased cell death.When the ultrasonic intensity was 1.2 W/cm2,the transfection efficiency and mortality were better than the other subgroups of group A.When the duty cycle was 20％,the transfection efficiency and mortality rate were better than the other subgroups of group B.Continue to increase the exposure time over 90 s was not statistically significant (P ＞0.05) for transfection efficiency and cell mortality.Conclusions The ultrasound-targeted microbubble destruction is an efficient method for gene delivery in vitro.The transfection efficiency vary greatly under different parameters,so optimization parameters is conducive to the promotion of gene transfection.

OBJECTIVETo observe the effect of purgation and detoxification therapy on gastrointestinal dysfunction of critically ill patients undergoing abdominal surgery.

METHODSTotally 56 inpatients with severe gastrointestinal dysfunction after abdominal surgery at ICU of Guangdong Provincial Hospital of Traditional Chinese Medicine were assigned to the treatment group and the control group, 28 in each group. All patients received routine Western medical treatment. Patients in the treatment group additionally took Modified Huanglian Jiedu Decoction (MHJD) and received electroacupuncture (EA) for 7 days. The first exhaust time, defecation time, scores for gastrointestinal dysfunction, mechanical ventilation time, ICU hospitalization time, and 28-day fatality rate were observed. Furthermore, serum levels of diamine oxidase (DAO) and D-lactic acid were detected at day 1, 3, and 7 after treatment.

RESULTSThe first exhaust time and the first defecation time in the treatment group were ahead of schedule, when compared with those of the control group (P <0. 05). Scores for gastrointestinal dysfunction, mechanical ventilation time, serum levels of DAO obviously decreased in the treatment group (P <0. 05). There was no statistical difference in serum levels of D-lactic acid, ICU stay time, the incidence of pulmonary infection, and 28-day mortality between the two groups (P >0. 05). Results of Logistic analysis showed that scores for gastrointestinal dysfunction were related with the incidence of pulmonary infection (P <0. 05).

CONCLUSIONMHJD combined EA could promote the recovery of gastrointestinal function in critically ill patients after abdominal surgery via improving intestinal barrier function, which was benefit for shortening mechanical ventilation time.

Critical Illness ; Defecation ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Electroacupuncture ; Gastrointestinal Diseases ; drug therapy ; Humans ; Lactic Acid ; Medicine, Chinese Traditional

OBJECTIVETo investigate the changes of intestinal microflora in two rat models of Pi-deficiency syndrome (PDS) to study the effect of Sijunzi Decoction (SJZD) on intestinal microflora alteration (IMA).

METHODSWistar rats allocated in Group A and B were established into two kinds of PDS models by Folium Sennae and Radix et Rhizoma Rhei respectively. Fecal samples were collected from the model rats at three phases, i. e. before modeling, after modeling and after treated with SJZD. Total DNA of microbial communities in them was extracted and amplified with ERIC-PCR to generate fingerprints for reflect the intestinal microflora configuration. Shannon-Weaver index (H') was used to analyze the diversity of rats' intestinal microflora, Sorenson pairwise similarity coefficient (Cs) was used to compare the similarity of different ERIC-PCR fingerprints, and Student's t test was used for analyzing the statistical significance of their differences.

RESULTSThe intestinal microflora in both groups obviously changed after modeling and recovered to the primary level after treated with SJZD. The H' of the ERIC-PCR fingerprints at the three phases was 2.20 +/- 0.12, 2.00 +/- 0.10 and 2.27 +/- 0.10 in Group A, and 2.09 +/- 0.02, 1.88 +/- 0.10 and 1.94 +/- 0.10 in Group B respectively, both showing significant differences among different phases (P<0.05). Accumulative curves of Cs showed that the proportion of Cs accounting for less than 60% was 0 in Group A and 19% in Group B before modeling, 75% and 52% for them respectively after modeling, and 4% and 24% after rehabilitation with SJZD.

CONCLUSIONThe H' of intestinal microflora in the two kinds of PDS models were significantly lowered, and could be recovered to the normal level by SJZD treatment (P<0.05). ERIC-PCR fingerprints can be used as a bio-criterion for evaluating intestinal microflora in patients with PDS.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Intestines ; drug effects ; microbiology ; Male ; Rats ; Rats, Wistar ; Syndrome

OBJECTIVETo analyze the status and characteristics of dental manpower in the center for disease controls (CDC) in Sichuan Province and to provide more evidence for strengthening the oral healthcare workforce in the CDC system.

METHODSA mass survey on dental manpower was made in CDCs in Sichuan Province through questionnaire investigation. Data were collected and entered with the Epidemiological Dynamic Data Collection (EDDC) platform and analyzed with SPSS 13.0 software.

RESULTSSichuan Province had 0.15 hospitals providing oral health services and 0.38 dentists on average per 10,000 people. About 65.53% (135/206) of the CDCs had one department responsible for the oral health service. However, oral health care personnel comprised only 2.23% (237/10,624) of the personnel of the whole CDC system. About 64.67% (119/184) of county CDCs and 47.62% (10/21) of city CDCs knew well the dental health status of local residents. Less than 5% of the CDCs used the data and assisted in the policy making of public health administrators.

CONCLUSIONThe dental care personal deficit exists in the CDC system in Sichuan Province. The distribution and composition of dental manpower are not reasonable. The oral health service ability of CDCs in Sichuan Province should be strengthened and improved.

China ; Dental Care ; statistics & numerical data ; Dentists ; Humans ; Oral Health ; Surveys and Questionnaires

BACKGROUND:Bone marrow mesenchymal stem cel s are considered to have good proliferation and differentiation potentials. Sox9 is a transcription factor that is essential for chondrogenesis and has been termed as a“master regulator”of the chondrocyte phenotype. OBJECTIVE:To study the therapeutic effects of Sox9-transfected bone marrow mesenchymal stem cel s on knee osteoarthritis. METHODS:The bone marrow mesenchymal stem cel s were transfected with Lenti-Sox9-EGFP in vitro. The model of murine knee osteoarthritis was established by cutting off the anterior cruciate ligament. Thirty model mice were randomly divided into three groups, as normal saline group, bone mesenchymal stem cel group and Sox9-transfected bone mesenchymal stem cel group. 0.1 mL of normal saline, 0.1 mL of normal saline containing non-transfected bone marrow mesenchymal cel s (non-transfected group), or 0.1 mL of normal saline containing Sox9-transfected bone marrow mesenchymal cel s (Sox9-transfected group) was injected into the knee joint cavity of mice in the corresponding group, respectively. After 4, 8, 12 weeks, the repair of articular cartilage lesions was evaluated by toluidine blue and immunohistochemical staining. RESULTS AND CONCLUSION:The lesions of articular cartilage were more serious in the normal saline group, compared with the other two groups, and the difference became more obvious over time. Damaged articular cartilage was improved in the non-transfected group, but the improvement was less than that in the Sox9-transfected group. Immunohistochemistry staining revealed that in the Sox9-transfected group, the positive type II col agen expression was stronger than that in the other two groups, but this positive expression was decreased over time in al the three groups. These results suggest that Sox9-transfected bone marrow mesenchymal stem cel s promote the repair of damaged cartilage in mice with knee osteoarthritis.

ObjectiveTo investigate the differences of whole blood chemical elements and urinary fluorine between patients with endemic fluomsis and patients without endemic fluorosis,and to find out the elements associated with endemic fluorosis and further lay a theoretical basis for clarify the pathogenesis of the disease.MethodsUsing case-control study,100 children aged 8 - 12 with dental fluorosis in Wushan and Fengjie counties of Chongqing from December 2010 to February 2011,and 30 adults with skeletal fluorosis were enrolled as case group; 100 children aged 8 - 12 without dental fluorosis and 30 adults without skeletal fluorosis were enrolled as internal control group; and 50 children without dental fluorosis and 30 healthy adults were selected as external control group in non-epidemic areas in Yubei district.Whole blood copper,zinc,calcium,magnesium,iron and urinary fluorine of all subjects were determined,and differences of these indexes were compared between groups.ResultsThe levels of copper,zinc,calcium,magnesium,iron and urinary fluorine of children in the case group were (30.08 ± 2.83),(74.04 ± 9.75)μmol/L,(1.65 ± 0.29),(1.37 ± 0.17),(6.79 ± 1.27)mmol/L,and (0.73 ±0.37)mg/L,respectively; the levels of these elements of children in internal control group were (28.65 ± 3.96),(72.83 ± 11.35)μmol/L,(1.62 ± 0.27),(1.36 ± 0.18),(6.73 ± 1.22)mmol/L,and (0.48 ± 0.21)mg/L,respectively; in external control group were (32.03 ± 2.99),(77.78 ± 10.85)μmol/L,(1.41 ± 0.11),(1.43 ± 0.13),(7.66 ±0.55)mmol/L,and (0.49 ± 0.26)mg/L,respectively(all P＜ 0.05),the comparison between any two groups indicated the levels of copper,zinc,magnesium,iron of the case group were lower than that of external control group,urinary fluorine was higher than that of internal and external control groups(all P ＜ 0.05).The levels of copper,zinc,calcium,magnesium,iron and urinary fluorine of adult case were (26.93 ± 4.37),(95.89 ± 12.45)μmol/L,(1.50 ± 1.76),(1.56 ± 1.96),(8.15 ± 1.00)mmol/L,and (2.17 ± 0.99)mg/L; internal control group were (26.26 ±4.96),(94.86 ± 12.18)μmol/L,(1.57 ± 0.12),(1.46 ± 0.16),(7.64 ± 1.00)mmol/L,and (1.44 ± 1.22)mg/L;external control group were (26.20 ± 2.96),(96.52 ± 11.11)μmol/L,(1.48 ± 0.14),(1.45 ± 0.16),(7.81 ±0.91 )mmol/L,and (0.55 ± 0.21 )mg/L,respectively.The levels of magnesium,iron and urinary fluorine of case group were higher than that of internal control group,magnesium and urinary fluorine were higher than that of external control group(all P ＜ 0.05).ConclusionsIn vivo anti-fluorine elements are deficient in the areas with endemic fluorosis.Other chemical elements,the environment and genetic factors may be related to the pathogenesisof the disease,which needs a further comprehensive analysis.

Objective To investigate the correlations of plasma homocysteine(Hcy)level and apolipoprotein E gene polymorphism with Alzheimer' s disease(AD)and mild cognitive impairment (MCI).Methods A case-control study in 66 AD patients(AD group),64 MCI patients(MCI group) and 54 healthy controls(control group)was conducted.Plasma Hcy level and ApoE polymorphism were determined and analyzed.Results Plasma Hcy levels were significantly higher in AD and MCI groups than in control subjects(both P＜0.001).AD patients also showed increased plasma Hcy levels as compared with MCI patients(P＜0.001).Logistic regression analysis indicated that the increased plasma Hcy level was a risk factor for AD and MCI(OR＝ 1.435 and 1.312,both P＜0.001).ApoE ε3/3 was the most common genotype in AD,MCI and control groups,and ε3/4 and ε4/4 genotypes were more common in AD group and MCI group than in control group(both P＜0.05).The ε4 allele frequency of ApoE was 24.2％ and 23.4％ in AD or MCI group respectively,and 6.5％ in control group(AD or MCI vs.control,P＜0.05).The analysis by multiplicative interaction model showed that the odd ratio for MCI was 23.3 in patients with only hyperhomocysteinemia(Hhcy,Hcy＞ 15 μmol/L),12.6 in patients with carrying ε4 allele,and 46.7 in patients with both Hhcy and carrying ε4 allele,which indicated that there was interaction between hyperhomocysteinemia and carrying e4 allele.Conclusions Hyperhomocysteinemia and ApoE ε4 allele are correlated with dementia and also have additive interactions.

To characterize the CD45RA+and CD45RO+T lymphocyte subsets in the peripheral blood of patients with cGVHD induced by allogeneic haematopoietic stem cell transplantation ( allo-HSCT ) and to explore their relations with the disease.Methods:The peripheral blood was collected from 64 patients after allo-HSCT,including 21 non-cGVHD patients,15 light grade cGVHD patients,18 mild grade cGVHD patients and 10 severe grade cGVHD patients,then CD4+CD45RA+,CD4+CD45RO+, CD8+CD45RA+and CD8+CD45RO+T lymphocyte subsets were detected by flow cytometry ( FCM).Results: Compared with the control,the percent of CD4+CD45RA+T lymphocyte in patients with light,mild and severe grade cGVHD decreased markedly (P<0.05),the percent of CD4+CD45RO+T lymphocyte increased markedly (P<0.05).But there were not obviously change in the patient with different grade cGVHD.The percent of CD8+CD45RA+,CD8+CD45RO+T lymphocyte did not change obviously.Conclusion:CD4+CD45RA+and CD4+CD45RO+may play an important role in the pathogenesis of cGVHD.