Objective To establish an ICP-MS method for the determination of heavy metals,including copper(Cu),arsenium(As),cadmium(Cd) and plumbum(Pb),in five kinds of Chinese medicinal material.Methods The samples were digested by closed-vessel microwave.The four heavy metals were directly analyzed by ICP-MS,with elements 72Ge,115In,and 209Bi as the internal standards.Results For all of the analyzed heavy metals,the correlative coefficient of the calibration curves was in the range of 0.999 5～ 0.999 7.The recovery rates were in the range of 92.3 % ～ 108.0 %,and RSD were in the range of 2.4 % ～ 5.8 %.Conclusion The method is convenient,rapid,accurate and can be applied to determine four heavy metals,including Cu,As,Cd and Pb,in Chinese medicinal material.

Objective To determine the conditioning regimen suitable for mice allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods Twelve BALB/c mice were randomly divided into 2 equal groups to undergo X-ray irradiation by linear accelerator at the dose of 7.0 Gy (pure X-ray group) or 60Co source irradiation at the dose of 7.0 Gy (pure γ-ray group).Thirty mice were randomly divided into 2 equal groups to undergo X-ray irradiation and then infusion of bone marrow from donor mice via caudal vein (X-ray + transplantation group) or γ-ray and then infusion of bone marrow via caudal vein (γ-ray + transplahtation group).3,5,7,10,15,20,and 30 d later peripheral blood samples were collected to calculate the number of white blood cells (WBCs) and detect the chimeric rates of lymphocytes by flow cytometry.5,10,and 20 d after irradiation 15 mice were killed with their lung,liver,small intestine,spleen,and femurs taken out to undergo pathological examination.Results The survival rates during the period 5-15 days of the γ-ray + transplantation group were all significantly higher than those of the X-ray + transplantation group.The pathological changes of organs of the X-ray +transplantation group were all more severe than those of the γ-ray + transplantation group.Since the fifth day after transplantation cells originating from the donor began to appear in the peripheral blood.The chimeric rate of the γ-ray + transplantation group 10 days after transplantation was (95.53± 2.57) %.The chimeric rates 5,10,and 20 days after transplantation of the γ-ray + transplantation group were all significantly higher than those of the X-ray + transplantation group (t = 15.263,3.256,P < 0.05).The WBC count of both irradiation groups decreased to the lowest level 5 d later and began to increase 10 days after transplantation and the WBC counts of the γ-ray + transplantation group 10 and 20 days aftertransplantation were both significantly higher than those of the X-ray + transplantation group (t = 3.624,6.695 ,P < 0.05).The chimeric rats of the peripheral lymphocytes 10 and 20 days after transplantation of the γ-ray + transplantation group were both significantly higher than those of the X-ray + transplantation group (t = 12.317,8.295,P < 0.05).The homogeneity rate of transplantation of the γ-ray +transplantation group was better than that of the X-ray + transplantation group.Conclusions As a conditioning regimen in allogeneic hematopoietic stem cell transplantation γ-ray irradiation causes milder injury and accelerated reconstitution of hematopoiesis and immunity,in comparison with X-ray irradiation.

Objective To study the relationship between graft-versus-host disease (GVHD) and endothelium injury following hematopoietic stem cells transplantation in mice. Methods C57BL/6 mice as donors and Balb/c mice as recipients were randomly divided into 4 groups: control group, bone marrow transplantation group, GVHD group, GVHD mitigation group. The clinical manifestations,circulating endothelial cells and tissue pathological changes were observed at different time points after transplantation. Results No manifestations of GVHD were found in each group at the day 5, while those were found in GVHD group at the day 9 and all died within 15 days. The counts of endothelial cells in peripheral blood showed no significant difference at the day 5 between GVHD group (7. 34 ±1.26 cells/μl) and bone marrow transplantation group (11.51 ± 7. 40 cells/μl) or GVHD mitigation group (7. 36 ± 0. 16 cells/μl), while among three groups there was statistically significant difference at the day 9 (GVHD group: 153. 64 ± 35. 35 cells/μl vs bone marrow transplantation group: 10. 49 ±5. 61 cells/μl and GVHD mitigation group: 47. 82 ± 4. 69 cells/μl). The scores of pathological aGVHD had no significant difference at the day 5 between GVHD group (4. 33± 1. 53) and bone marrow transplantation group (3. 33 ± 0. 58) or GVHD mitigation group (4. 00 ± 1.73), while among three groups there was statistically significant difference at the day 9 (GVHD group: 10. 0 vs bone marrow transplantation group: 3. 33 ± 1.15 or GVHD mitigation group: 4. 33 ± 0. 58) and at the day 14 (GVHD group: 10. 33 ± 2. 58 vs bone marrow transplantation group: 2. 33 ± 1.25 or GVHD mitigation group 3. 33 ± 1.15). Conclusion Occurrence of GVHD causes endothelial damage again and injured endothelium worsens the GVHD.

Objective To explore a proper dose of endothelial progenitor cells (EPCs)administration that can achieve optimal hematopoietic improving effectiveness in a murine allogeneic hernatopoietic stem cell transplantation (allo-HSCT) model.Methods Female Balb/c mice were lethally irradiated with 60Co source,and then were injected intravenously with 5 106 bone marrow cells from C57BL/6 mice (bone marrow transplantation group).In co-transfer experiments,5 × 104,1 ×105,5 × 105 or 1 × 106 donor EPCs (EPCs treated groups) were injected simultaneously with bone marrow cells.The recipients were monitored for survival,peripheral white blood cells,hematopoietic stem cells (HSCs) and bone marrow histology.Results Compared with bone marrow transplantation group,all EPCs treated groups had accelerated recovery of peripheral white blood cells (P＜0.05),platelets (P＜0.05) and HSCs (P＜0.05).When infused with less than 5 × 105 EPCs,these effective hernatopoietic improving phenomena showed a positive correlation with the administrated doses of EPCs.However,when infused with 1 × 106 EPCs,the mice showed lower survival rate (P＜0.05)and slower recovery of peripheral white blood cells (P＜0.05),platelets (P＜0.05) and HSCs (P＜0.05) than 5 × 105 EPCs treated grpup.Bone marrow histopathology analysis confirmed the above findings.Conclusion Co-transfer with donor EPCs can improve survival rate and hematopoietic reconstitution of recipient mice in allo-HSCT,and 5 × 105 EPCs should be a proper dose to achieve the best effectiveness.

OBJECTIVETo investigate the frequency of CYP2C19 polymorphisms involved in clopidogrel metabolism in Fujian Han population.

METHODSFrequencies of CYP2C19* 2, CYP2C19*3 and CYP2C19*17 in 1001 unrelated Fujian Han volunteers were determined with polymerase chain reaction-restriction fragment length polymorphism and direct sequencing method.

RESULTSThe frequencies of CYP2C19*2, *3 and *17 were 32.4%, 5.8% and 0.4%, respectively. According to genotyping results, intermediate metabolizers (CYP2C19 *1/*2 or *1/*3) and poor metabolizers (CYP2C19 *2/*2 and *2/*3) respectively accounted for 47.95% and 13.99% of all subjects. Above frequencies were similar to those of Japan, Korea, Singapore, Malaysia, Thailand and Chinese Dai, Mongolian,Li and Hui ethnics (P>0.05), but were significantly different from those of Chinese Kazakh and Uygur ethnics, and people from Iran, Russia, Italy, Poland, Norway, Canada native Indians, Bolivia, Egypt or Tanzania (P<0.05).

CONCLUSIONEthnic/regional diversity exist with regard to the prevalence of CYP2C19 polymorphisms. No significant difference were found between Fujian Han Chinese and Dai, Mongolian, Li and Hui from China or other populations from East and Southeast Asia, but higher frequencies of intermediate metabolizers and poor metabolizers compared with populations of Kazakh and Uygur in China, and people from Europe, South America and Africa.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Aryl Hydrocarbon Hydroxylases ; genetics ; China ; Cytochrome P-450 CYP2C19 ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Middle Aged ; Polymorphism, Genetic ; Ticlopidine ; analogs & derivatives ; metabolism ; Young Adult

OBJECTIVETo detect the inhibitory effect of all-trans retinoic acid(ATRA) on breast cancer stem cells (CSCs).

METHODSThe inhibitory effect of ATRA on MCF-7 and SK-BR-3 cell lines was analyzed using a Cell Counting Kit-8 (CCK-8). The proportion of CD44(+)CD24(-) tumor cells of the two cell lines were measured before and after the ATRA treatment, and the role of ATRA in the regulation of CSC self-renewing ability was evaluated with a tumor sphere assay. The tumor spheres were grown in an adherent culture to evaluate the ATRA-induced differentiation of breast cancer stem cells.

RESULTSATRA effectively inhibited the unsorted cells and stem cells, but the CSCs were more sensitive to ATRA. At a concentration of 10(-6) mol/L, the inhibitory rate of MCF-7 unsorted cells and stem cells were (8.66 ± 1.06)% and (21.09 ± 3.25)%, respectively (P = 0.004). For SK-BR-3 cells, the rates were (39.19 ± 1.47)% and (51.22 ± 2.80)%, respectively (P = 0.005). The self-renewing ability of the CSCs was impaired by ATRA at a concentration of 10(-6) mol/L. The rate of MCF-7 and SK-BR-3 stem cells to form tumor sphere was 5.2% (5/96) and 13.5% (13/96), respectively. For the control group, it was 86.5% (83/96) and 93.8% (90/96), respectively (P < 0.001). ATRA also promoted the CD44(+)CD24(-) subpopulation to differentiate. SK-BR-3 stem cells were grown in an adherent culture. After using ATRA, the proportion of CD44(+)CD24(-) cells was (48.1 ± 2.5)% and that of the control group was (86.6 ± 2.5)% (P < 0.001).

CONCLUSIONSATRA effectively inhibits breast NCSCs and CSCs, but CSCs are more sensitive to ATRA. ATRA impairs the self-renewing ability of CSCs and promotes CSCs to differentiate.

Antineoplastic Agents ; pharmacology ; Breast Neoplasms ; metabolism ; pathology ; CD24 Antigen ; metabolism ; Cell Differentiation ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Female ; Humans ; Hyaluronan Receptors ; metabolism ; Neoplastic Stem Cells ; cytology ; drug effects ; Tretinoin ; pharmacology

Oncolytic herpes simplex virus (HSV) can replicate in and kill cancer cells without harming normal tissue. G47delta is a third-generation HSV vector. In this study, the therapeutic effects of G47delta on human nasopharyngeal carcinoma (NPC) were determined in vitro and in vivo. The human NPC cell lines CNE-2 and SUNE-1, primary normal nasopharyngeal epithelial cells (NPECs), and immortalized nasopharyngeal cells NP-69 and NPEC2/Bmi1 were infected with G47delta at different multiplicities of infection (MOIs). The survival of infected cells was observed daily. Two subcutaneous models of NPC were established with CNE-2 and SUNE-1 in Balb/c nude mice. G47delta or virus buffer as control was injected into the subcutaneous tumors. Tumor size was measured twice a week, and animals were euthanized when the diameter of their tumors exceeded 18 mm or when the animals appeared moribund. For the NPC cell lines CNE-2 and SUNE-1, more than 85% and 95% of cells were killed on day 5 after G47delta infection at MOI = 0.01 and MOI = 0.1, respectively. Similar results were observed for an immortalized cell line NPEC2/Bmi-1. A moderate effect of G47delta was also found on another immortalized cell line NP-69, of which only 27.7% and 75.9% of cells were killed at MOI = 0.01 and MOI = 0.1, respectively. On the contrary, there was almost no effect observed on NPECs. The in vivo experiments showed that tumors in mice in the G47delta-treated group regressed completely, and the mice exhibited much longer survival time than those in the control groups. Our results suggest that the potential therapeutic effects of G47delta would be applicable for treatment of NPC patients in the future.
Animals ; Apoptosis ; Carcinoma ; Cell Line, Tumor ; Female ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Nasopharyngeal Neoplasms ; pathology ; therapy ; virology ; Oncolytic Virotherapy ; methods ; Oncolytic Viruses ; physiology ; Simplexvirus ; physiology ; Xenograft Model Antitumor Assays

OBJECTIVESTo investigate the transmission process of severe acute respiratory syndrome (SARS) and to evaluate the infectiveness of SARS patients in different periods of disease epidemics.

METHODSStandardized questionnaire was used to conduct case investigation and contact tracing by combining the field investigation and telephone interview. Transmission process, infectivity, transmission chain and contact history of SARS were studied through data analyses.

RESULTSOn 25th March 2003, a 91 year old man was admitted to Hospital J in Beijing with stroke and fever. He died on 30th March. From 31st March, there was an outbreak of SARS among his contacts in the family and in the hospital he was admitted to. Contacts would include his relatives, other co-patients and health care workers in the Hospital J. Chinese Field Epidemiology Training Program trainees conducted an investigation of the outbreak. Among the 207 contacts of the index cases through different generations, there were 36 cases of SARS (attack rate 17%) patients with one death. There were 12 cases having directly contact with the index case and 13 cases with one secondary case. The transmission chains of this outbreak could clearly be depicted. All the cases had close contacts during the symptomatic period of their index patients. Among the relatives, 85% of the cases had 3 - 5-day contact with their index patients after the onset of the illnesses. There was no significant difference between the two attack rates-70% for whose who had contact with the patient before and after illness onset) and 67% for those who only had contact after the onset of the illness. Out of the 44 social acquaintances and 38 of the family members who had contacts with the index patients during the incubation period, no one was found ill. Among the close contacts at the hospital who had no protection when providing care to the patient, the attack rate was found over 80%.

CONCLUSIONSAll the secondary cases of this outbreak had a history of direct and close contacts to the index patients after the onset of the illness. There was no evidence indicating that SARS cases were infectious during their incubation period.

Aged ; Aged, 80 and over ; China ; epidemiology ; Humans ; Infectious Disease Transmission, Patient-to-Professional ; Male ; Severe Acute Respiratory Syndrome ; epidemiology ; prevention & control ; transmission

Objective:To discuss the topical action characteristics of the biological transmission of moxibustion heat via temperature collection and numerical modeling.Methods:Temperature of moxibustion was measured at multiple points at a distance of 3 cm to obtain the moxibustion temperature field nephograms by the high-accuracy temperature measure array.Finite element analysis was used to imitate the three-dimensional dynamic distribution of temperature in acupoint tissues.Results:Through numerical analysis,the one-dimensional,two-dimensional and three-dimensional distributions of temperature in human acupoint tissues at 5 min of moxibustion were established.The result showed that moxibustion heat mainly transmitted from the surface of the tissue to the internal,and the influence of moxibustion heat decreased with the depth of the tissue.The analysis of the nephograms of acupoint tissue temperature at 5,10,15 and 20 min of moxibustion showed that with the increase of the moxibustion time,the temperature in acupoint tissues constantly rose,and the transmission depth of moxibustion heat also further expanded inside acupoint.Conclusion:By establishing the three-dimensional dynamic model of heat transmission inside acupoint tissues with the biological parameters of human tissues and the temperature values obtained,this study used finite element analysis software ANSYS 14.0 and discovered the rules in the transmission of heat in body tissues during moxibustion,and the features in moxibustion heat transmission (from the proximal to the distant) and heat penetration (from the surface to the internal).This study provides theoretical and experimental support for the application of moxibustion in clinical practice.

OBJECTIVETo study the role of O(6)-methylguanine-DNA methyltransferase (hMGMT) in the development of human lung cancer.

METHODSReverse transcription-polymerase chain reaction (RT-PCR) method was applied to measure hMGMT mRNA expression in 150 lung cancer specimens, 40 normal lung tissues, and in the peripheral mononuclear blood cells from 50 lung cancer cases and 50 normal controls. The protein expressions of p53, C-MYC and K-RAS were assessed by immuno-histochemistry. The effects of some exposure factors on the expression of hMGMT gene were analyzed. The relationships between hMGMT gene and cancer related genes p53, C-MYC and K-RAS were investigated.

RESULTSThe mRNA of hMGMT was low or absent in 49 of 150 (32.7%) lung cancer specimens, whereas 2 of 40 (5%) normal lung tissues had reduced the levels of hMGMT mRNA. The low expression of hMGMT seemed to be a risk factor of lung cancer, with a OR of 9.22 (2.05-57.65). Reduced expression levels of hMGMT mRNA were observed in 10 of 50 (20%) lung cancer patients' peripheral mononuclear blood cells, and 2 of 50 (4%) blood cells among normal controls. When investigating the exposure factors which affecting the expression of hMGMT gene, we noticed that smoking was suppressing the expression of hMGMT gene. Interestingly, over-expression of K-RAS oncogene was significantly correlated with low expression of hMGMT (P < 0.05). However, the expressions of p53 and C-myc were not correlated with the status of hMGMT gene.

CONCLUSIONhMGMT might play an important role in the development of human lung cancer. Low expression of hMGMT gene seemed to be a risk factor for lung cancer which could be used as a valuable biomarker on susceptibility of human lung cancers.

Adult ; Aged ; Biomarkers, Tumor ; Carcinoma, Squamous Cell ; enzymology ; genetics ; China ; epidemiology ; DNA Repair ; genetics ; Female ; Genes, ras ; genetics ; Humans ; Lung Neoplasms ; enzymology ; genetics ; Male ; Middle Aged ; O(6)-Methylguanine-DNA Methyltransferase ; biosynthesis ; genetics ; metabolism ; Point Mutation ; RNA, Messenger ; biosynthesis ; genetics ; Smoking ; adverse effects ; ras Proteins ; biosynthesis ; genetics