To explore the new method of discriminating Cistanche deserticola, Cynomorium songaricum and Orobanche pycnostachya by using PCR amplification of specific alleles. 30 samples of the different C. deserticola, 21 samples of C. songaricum and O. pycnostachya were collected. The total DNA of the samples were extracted, the ITS sequences from C. deserticola, C. songaricum and O. pycnostachya were amplified by PCR and sequenced unidirectionally. These sequences were aligned by using ClustulW. Specific primer was designed according to the ITS sequences of specific alleles, and PCR reaction system was optimized. Additionally, compare with the identification of specific PCR method and DNA sequence analysis method. The result showed that the 331 bp identification band for C. deserticola and the adulterants not amplified bands by a single PCR reaction, which showed good identification ability to the three species. PCR amplification of specific alleles can be used to identify C. deserticola, C. songaricum and O. pycnostachya successfully.
Alleles ; Cistanche ; classification ; genetics ; DNA Primers ; genetics ; DNA, Intergenic ; genetics ; DNA, Plant ; genetics ; Drug Contamination ; prevention & control ; Phylogeny ; Polymerase Chain Reaction ; methods

Objective To evaluate the use of double balloon endoscopy(DBE) in diagnosis of ulcerative lesions in small intestine.Methods Data of patients diagnosed as small intestinal ulcer under DBE during September 2003 and December 2007 at Nanfang Hospital were analyzed retrospectively.Results Ulcer in small intestine was detected by DBE in a total of 62 patients,including 48 males and 14 females,aging from 10 to 71 years old( mean 43.9 yr).The main clinical manifestations consisted of small intestinal hemorrhage(38/62,61.3%),abdominal pain(16/62,25.8%),abdominal distention(5/62,8.1%),loss of weight(2/62,3.2%),and diarrhea(1/62,1.6%).The ulcers were diagnosed endoscopically as Crohn's disease(CD) in 53 cases(85.5%),drug induced lesions in 4(6.5%),nonspecific chronic inflammation in 2(3.2%),lymphoma in 2(3.2%) and tuberculosis in 1(1.6%).They were all microscopically diagnosed as chronic inflammation.Of the 62 patients,32(51.6%) underwent surgery.In 30 cases of CD diagnosed by DBE,22 were confirmed by post-surgery pathology(malignant cells were found in 3),while in the other 8 cases,4 were diagnosed as lymphoma,3 as Behcet's disease and 1 as tuberculosis.Meanwhile,the 1 case of tuberculosis and 1 lymphoma diagnosed by DBE were confirmed as CD after operation.The overall accurate diagnosis rate of small intestinal ulcerative lesions by DBE was 68.8%(22/32).Conclusion DBE is valuable in diagnosis of ulcerative lesions in small intestine,but surgery should be included into consideration to confirm the diagnosis when necessary.

Objective To explore the effect of nuclear factor-kappaB(NF-?B) on cardiac myocyte apoptosis and understand the molecular mechanism of decrease of heart function in septic shock rats.Methods Twenty Wistar rats were randomly divided into the septic shock group and the normal control group.The septic shock group(n=10) were fixed with anaesthesia and made into 1.5 cm slices just along the abdomen midline.Then the roots of appendices were returned to the belly cavity after being ligated and punctured 4 times with the No.14 pinhead.After that,the slices were sewn up layer by layer.After 12 hours,the rats were all sacrificed,the blood taken and the serum separated.In the end,the heart specimens of the septic shock group were set aside.Meanwhile,the normal control group were dealt with in the same way except that they were not subjected to cecal ligation and puncture.Then NF-?B protein levels in cardiac tissue and the index of cardiac myocyte apoptosis were measured.Results NF-?B protein levels in the septic shock group(9 cases were strong masculine gender,1 case was middle masculine gender)elevated significantly compared with the normal control group(8 cases were negative gender,2 cases were weak masculine gender) in cardiac tissue(P

AIM AND METHODSThe distribution of ErbB2 in mouse testis, epididymidis, ovaries, oocyte-cumulus cells-complexes in oviducts and sperms was investigated immunohistochemically. To study the effect of c-erbB2 on mouse fertilization in vitro, various concentrations of c-erbB2 antisense oligonucleotides (c-erbB2 ASODNs) were incubated with sperms and oocyte-cumulus cells-complexes during fertilization in vitro. To explore possible mechanisms involved in fertilization, the relationship between c-erbB2 ASODNs and GABA, or dbcAMP, or verapamil during fertilization in vitro was also observed.

RESULTSErbB2 oncoprotein was observed in epithelial cells in epididymis, sperms and cumulus cells. C-erbB2 ASODNs inhibited the rate of fertilization in vitro in a dose-dependent way. The fertilization rate of the control group, low (5 micromol/L), medium (10 micromol/L), high (20 micromol/L) concentration c-erbB2 ASODNs group, and nonsense at oligonucleotides group (20 micromol/L) was 38.3%, 19.6%, 10.7%, 5.0%, and 33.8% respectively. Integral optical density immunostaining of ErbB2 in sperms was notably reduced. Medium and high concentration of c-erbB2 ASODNs notably inhibited cumulus cells adhering to inner wall of Petri dish. Treated alone with GABA or dbcAMP, the rate of fertilization was increased. Both GABA and dbcAMP partially inversed the ASODNs inhibition effect on fertilization rate, but neither of them showed significant effect on sperm integral optical density of ErbB2 immunostaining. In contrast, verapamil inhibited fertilization rate. Co-treated with c-erbB2 ASODN, verapamil showed synergic inhibiting effect on fertilization with c-erbB2 ASODN. Verapamil also inhibited the expression of c-erbB2 in sperms.

CONCLUSIONIt is suggested that c-erbB2 is closely correlated with fertilization. Ca2+ may inhibit fertilization in vitro through regulation the expression of c-erbB2 gene in sperm cells, while both of GABA and dbcAMP may affect the process of fertilization through the way other than c-erbB2 expression in sperm cells.

Animals ; Bucladesine ; pharmacology ; Calcium ; physiology ; Epididymis ; physiology ; Female ; Fertilization ; physiology ; Fertilization in Vitro ; Male ; Mice ; Mice, Inbred Strains ; Oligonucleotides, Antisense ; pharmacology ; Oocytes ; physiology ; Ovarian Follicle ; physiology ; Receptor, ErbB-2 ; physiology ; Sperm-Ovum Interactions ; Verapamil ; pharmacology ; gamma-Aminobutyric Acid ; pharmacology

OBJECTIVETo study the characteristics of EV71 JN200804 strain infection in one-day old BALBI c mjce and to establish a animal model of EV71 infection , and to provide information and technical support for the evaluation of the EV71 vaccine and antiviral medicine.

METHODSOne-day old BALBic mice were infected with EV71 JN200804 strain through oral( PO) ,intracranial(IC) ,intraperitoneal (IP), intramuscular (IM) routes, respectively. All mice were sacrificed at paralysis of hind limbs and collected organs for viral isolation, RT -PCR and pathological examination, and the electrophysiology were detected before sacrifice.

RESULTSAll mice infected through IC, IP and IM routes were paralyzed in hind limbs at 4-5 days and died at 7 days about, the hypokinesia and lethargy of mice were observed through PO routes. The viruses could be isolated and detected in the muscle from mice infected through IC, IP and IM routes and in the spinal cord through IC routes by viral isolation and RT-PCR. The neurogenic and myogenic disorders were detected by electromyography. Histopathologically, the varied pathologic changes were observed in the mouse cerebellum , spinal cord , muscle , heart, lung, liver and kidney.

CONCLUSIONEV71 JN200804 strain can infect one-day old BALBI c mice and induce paralysis of hind limbs, its animal infection model may apply to study of EV71 infection pathogenesis and antiviral medicine, and evaluation of the EV71 vaccine.

Animals ; Disease Models, Animal ; Enterovirus A, Human ; immunology ; physiology ; Enterovirus Infections ; immunology ; virology ; Mice ; Mice, Inbred BALB C

Objective To investigate the type 2 diabetes mellitus(T2DM)prevalence and related risk factors in adult population with obesity in Tianjin. Methods With stratified cluster randomized sampling, 2888 obese people with BMI≥28 kg/m2, aged 18 years old and over were selected from three urban and three rural regions of Tianjin, in 2006. Information on risk factors was collected with questionnaire through face-to-face interview by trained workers and data on fasting blood glucose(FBG)was collected at the same time. 2hrPPG was tested among the people who' s FBG ≥6.1 mmol/L at the hospital. Prevalence of T2DM was calculated and the distribution of T2DM in the described subgroups and the risk factors analyzed with SPSS software. Results The prevalence of T2DM in adult population with obesity was 11.74%, with females(13.90%)higher than males (8.75%). The prevalence rates of T2DM were statistically different among different groups, classified by age, education, occupation, district and BMI. Results from the univariate and multivariate logistic regression analysis showed that the risk factors of T2DM were age(OR=1.383, 95% CI: 1.254-1 .525)and sex(OR= 1.591,95% CI: 1.230-2.059)while the protective factor was fruit intake(OR=0.867, 95% CI: 0.774-0.971). Conclusion The prevalence of T2DM in adult with obesity was considered to be high. The distribution of T2DM in different subgroups and affecting factors of T2DM in obese adults were different from general population.

Objective To investigate the spiral CT features of gastric carcinoma in the invasion and metastasis and its correlation with the expression of phosphatase and tensin horology deleted on chormosome ten(PTEN)and basic fibroblast grouth factor(bFGF).Methods Spiral CT plain scan and triphasic enhanced scans were performed in 83 patients.The postoperative specimens were embedded with paraffin to obtain 5?m thickness tissues and stained with HE and immunohistochemistry.Spiral CT findings were compared with the expression of phosphatase and tensin horology deleted on chormosome ten(PTEN)and basic fibroblast growth factor(bFGF).Results(1)The accuracy of spiral CT in T and N staging of gastric carcinoma was 94.0%(78/83)and 89.2%(74/83),respectively.(2)The expression of PTEN was 47.0%(39/83)in gastric carcinoma.The expression of PTEN in T_(3.4)(40.8%,29/71)and N_(1+2) (38.3%,23/60)gastric carcinoma was significantly lower than that of T_2(10/12)and N_0(16/23)gastric carcinoma,respectively(X~2=7.439,P=0.006;X~2=6.511,P=0.011).(3)The expression of bFGF was 63.9%(53/83)in gastric carcinoma.The expression of bFGF in T(3.4)(70.4%,50/71)and N_(1+2) (71.7%,43/60)gastric carcinoma was significantly higher than that of T_2(3/12)and N_0(10/23)gastric carcinoma,respectively(X~2=7.314,P=0.007;X~2=5.724,P=0.017).(4)Both PTEN-positive expression and bFGF-positive expression were detected in 16 specimens.The expression of PTEN(41.0%, 16/39)was negatively correlated with that of bFGF(30.2%,16/53)(r=-0.447,P=0.000). Conclusion Spiral CT triphasic enhanced scans combined with biologic characteristics can improve diagnostic accuracy of gastric carcinoma in the invasion,metastasis and prognosis.

Background Qiang minority is minority groups of China with the special habits and customs and living condition. So whether the spectrum of disease and bacteria spectrum in conjunctiva are similar with Han nationality is worth paying attention. Objective Present survey was to obtain the data about bacterial species in conjunctival sac in Qiang minority population with the age 40 years old and more and the compare with matched Han nationality population. Methods This survey study was performed as the standardized training and protocol. A total of 212 eyes of 106 individuals from Qiang minority in Beichuan county and 640 eyes of 320 subjects from Han nationality in Mianyang city received questionnaire survey and ophthalmological examination. The secretion of the inferior palpebral conjunctival sac was embrocated and inoculated on blood plate for 48-72 hours. The bacteria was separated and identified. This study was approved by the Ethic Committee of Sichuan Provicial People' s Hospital. Orally informed consent was obtained before the medical procedure. Results All the examinee finished the survey and examination with a good compliance. No significant difference was found in the demography between these two groups of population. The multiple bacterial positive rate in conjunctival sac was 59. 4% in Qiang minority and that of Han people was 66. 3% with a considerably difference between them (χ2 = 2. 27,P = 0. 13). The multiple bacterial species were simultaneously detected in 26.2% in Qiang minority population and 11.88% Han people, showing evidently difference (χ2 = 106. 40, P = 0. 00 ) . The positive rate of corynbaccterium in conjunctival sac of Qiang minority was statistically lower than that of Han people (20. 7% versus 45. 0% ,χ2 =31. 75 ,P = 0. 00) ,but there was no statistical difference in the positive rate of staphylococcus epidemics between two groups (χ2 = 1. 89 ,P = 0. 17). Conclusion The bacteria positive rate in conjunctiva sac is resemble in the population over 40 years in both the Qiang minority and Han nationality. The simple bacterial species is found in majority people in two groups of subjects. The positive rate of multiple bacterial strains coexistence is more in the Qiang minority. The bacterial strains is different between Qiang minority and Han nationality.

OBJECTIVE: To investigate the biological properties of Caski cell lines induced by exposing to the space environment. METHODS: Caski cells were carried in "Shen Zhou IV" airship. After 7 days of spaceflight, cells survived and were monocoloned, and the experimental methods such as cell morphological observation, the cell proliferation assay, flow cytometry cell cycle analysis, the soft agar assay, and tumorigenesis assay were used to analyze cell growth characteristics and malignant phenotypes. RESULTS: Altogether 1440 strains subclonal cell lines were established and 4 strains were screened. Compared with the control group, mutated cells appeared to have multiple cell morphological changes. Strains numbered 44F10 and 17E3 were screened due to their increased cell proliferation and tumorigenesis, and their cell cycles were induced to progress from G(1) to S phase, while strains 48A9 and 31F2 were opposite to 44F10 and 17E3 in cytological events. The average population double time of ground nomal control group, ground simulant control group, strains numbered 44F10, 17E3, 48A9 and 31F2 groups were 56.54, 58.44, 52.96, 51.46, 101.76 and 88.47h, respectively; compared with the control group, the average double time of strains numbered 44F10 and 17E3 was decline, but with no statistical significance. However, compared with the control groups, the average double time of 48A9 and 31F2 was significant increased (P<0.05). The colony formation rates were 9.7%, 9.3%, 14.7%, 12.1%, 0 and 0.1%, respectively, and the difference between the ground control groups and the other groups was significant (P<0.01); 6 groups of above-mentioned Caski cells were inoculated subcutaneously in Babl/c nude mice respectively. Forty-seven days later, the formed tumors in the nude mice were statistically analyzed and tested. The average weight of tumors of the above-mentioned groups was 0.066, 0.066, 0.175, 0.249, 0.011 and 0.018g. The difference between the ground control groups and other groups was significant (P<0.05). CONCLUSION Spaceflight may affect the physiological characteristics of tumor cells and the variation is complicated.
Animals ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Female ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Neoplasms, Experimental ; pathology ; Space Flight ; Transplantation, Heterologous ; Uterine Cervical Neoplasms ; pathology ; Weightlessness

OBJECTIVETo observe changes in T cell subsets in children with sepsis and their prognosis, and to investigate the clinical significance of these changes in the occurrence and development of sepsis.

METHODSFifty children with severe sepsis and 150 children with general sepsis were enrolled as subjects, and 50 age-matched healthy children were included as controls. The percentages of CD3(+), CD4(+) and CD8(+) T cells in peripheral blood and CD4(+)/CD8(+) ratio were measured by flow cytometry. The pediatric critical illness score (PCIS) was calculated within 24 hours of admission.

RESULTSThe children with severe sepsis showed significantly lower percentages of CD3(+), CD4(+) and CD8(+) T cells CD4(+)/CD8(+) ratio and PCIS than the controls and children with general sepsis (P<0.01). Among the 200 cases of sepsis, the percentages of CD3(+), CD4(+) and CD8(+) T cells, CD4(+)/CD8(+) ratio and PCIS were significantly lower in the cured group than in the deceased group.

CONCLUSIONSChildren with sepsis have different degrees of cellular immunosuppression, and the degree of cellular immunosuppression is significantly correlated with the severity of the disease. Detection of T cell subsets in peripheral blood is of great significance for evaluating immune function and judging disease severity in children with sepsis.

CD4-CD8 Ratio ; Child, Preschool ; Female ; Humans ; Male ; Prognosis ; Sepsis ; immunology ; T-Lymphocyte Subsets ; immunology