The CO I gene sequences of Qianghuoyu, Pachytriton labiatus and Gehyra mutilata were achieved by PCR amplification and bi-directional sequencing. Furthermore, a pair of specific primers SJYW1 and SJYW2 in the non-conservative district were designed through sequence alignment. The PCR reaction condition was established by changing the annealing temperature and cycle numbers. The results showed that 350 bp DNA fragment was amplified from Qianghuoyu in PCR with annealed temperature at 54 °C and the cycle number was 25 cycles, whereas not any DNA fragment was amplified from P. labiatus and G. mutilata under the same reaction condition. This method is well-performed in the identification of Qianghuoyu for its excellent specificity and repeatability.
Animals ; Drug Contamination ; Medicine, Tibetan Traditional ; Polymerase Chain Reaction ; methods

OBJECTIVETo develop the chromatographic fingerprints of composite Folii Isatidis injection by HPLC.

METHODThe separation was performed on a Diamonsil C18 column with a mobile phase consisting of methanol-water-phosphoric acid as gradient eluent at the flow rate of 1.0 mL x min(-1). The UV detection was set at 254 nm.

RESULTA standard HPLC fingerprint procedure was developed for composite Folii Isatidis injection, with 20 common peaks and a similarity threshold of 9.0 established.

CONCLUSIONThis method was accurate, repeatable and useful for the quality control of composite Folii Isatidis injection.

Chromatography, High Pressure Liquid ; methods ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; isolation & purification ; Ginkgo biloba ; chemistry ; Injections ; Isatis ; chemistry ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results ; Uridine ; analysis

In order to explore the influence of purified donor regulatory T cells (T(Reg) cells) infused after allogeneic bone marrow transplantation (allo-BMT) on GVHD and hematopoietic reconstitution of mice, an allo-BMT model of C(57)BL/6-->BALB/c mice was established. CD4(+)CD25(+)T(Reg) cells were purified through bead-magnetic activated cells separated from donor mice peripheral blood. The recipient mice were randomly divided into three groups: CD4(+)CD25(+) T cells, CD4(+)CD25(-) T cells and RPMI 1640 culture medium. These cells and RPMI 1640 were infused into recipient mice by caudal veins at about 6 to 8 hours after allo-BMT respectively. Incidence of GVHD, pathological lesion of liver, spleen, small intestine, survival time and hematopoietic reconstitution in the recipients were observed after allo-BMT. The results showed that the time for WBC > 1.0 x 10(9)/L was (8.14 +/- 3.26) days, (17.62 +/- 5.71) days, (19.81 +/- 6.77) days and the time for Plt > 20.0 x 10(9)/L was (5.29 +/- 1.34) days, (8.97 +/- 3.44) days, (9.52 +/- 3.92) days in T(Reg) positive cell group, T(Reg) negative cell group and the blank control group respectively, and the recovery times of WBC and Plt in T(Reg) positive cell group were faster than that in T(Reg) negative cell group and the blank control group (P < 0.05). The scores of GVHD were (1.33 +/- 0.58), (1.80 +/- 0.27), (1.93 +/- 0.45) in three groups of mice at about 15 days after allo-BMT, respectively, the GVHD in T(Reg) positive cell group was slighter than that in T(Reg) negative cell group and the blank control group (P < 0.05). It was found that GVHD pathologic manifestations of the liver, spleen and small intestine in T(Reg) positive cell group were slighter in a certain extent than those in other two groups at about (25 - 30) days after allo-BMT. The mean survival time in three groups was (41.45 +/- 17.88) days, (18.75 +/- 14.39) days and (25.67 +/- 16.84) days after allo-BMT, respectively, which in the T(Reg) positive cell group was significantly longer than that in other two groups (P < 0.05). It is concluded that donor-T(Reg) cell infusion can mitigate the GVHD so as to reach hematopoietic reconstitution and prolong survival time after allo-BMT in mice.
Animals ; Female ; Graft vs Host Disease ; immunology ; prevention & control ; Hematopoietic Stem Cell Transplantation ; adverse effects ; methods ; Interleukin-2 Receptor alpha Subunit ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Random Allocation ; T-Lymphocytes, Regulatory ; immunology ; transplantation

Adult ; Coronary Aneurysm ; complications ; Endocarditis, Bacterial ; complications ; Fistula ; complications ; Humans ; Male

Objective To investigate the distribution of water-borne fluoride and the current situation of endemic fluorosis in Yan'an city in 2006, and to evaluate the effect of water defluoridation project by improving driking water quality. Method In 2006 in Yan'an city, 5 samples from water source were collected in each selected village that was chosen according to 5 directions of East, West, South, North, and Central. Meanwhile, 1 sample from water source, 1 sample from water processing factory and 2 tap water samples were collected from each water defluoridation project. Water fluoride was determined by spectrophotometric method, teeth and skeletal fluorosis examination were performed by Dean method and "national criteria of endemic skeletal fluorosis diagnosis of China" in children aged 8-12 year and adults, respectively if water fluoride level > 1.00 mg/L Results Of 726 water samples from 293 villages tested, samples from 25 villages had higher fluoride( > 1.00 mg/L), and these villages covered a population of 11 610 people and most of these people were in Wuqi and Yanchuan counties. Water fluoride ranged from 0.10 mg/L to 3.50 mg/L, with median being 0.59 mg/L. Of 100 water samples from 25 water defluoridation projects, only 1 sample exceeded the national criteria in Yanchuan, and Wuqi counties, respectively,with fluoride level being 1.85 mg/L and 1.60 mg/L, respectively, and population exposed was 3083 and 708, respectively, with water fluoride ranged 0.30 - 2.00 mg/L In the examination of 1281 children aged 8 - 12, we detected 238 cases of dental fluorosis, and the detection rate reached 18.58%; 13 900 adults were checked, and 375 cases were confirmed of skeletal fluorosis, a detection rate reached 2.70%. Conclusions Yan'an has a wide range of water with high fluoride and severe fluorosis people. The water defluoridation projects need to be further improved. The task of prevention of endemic fluorosis is still arduous, and we should speed up the implementation of comprehensive water defluoridation measures.

Fibronectin containing extra domain A (EDA+ FN),a functional glycoprotein participating in several cellular processes,correlates with chronic liver disease.Herein,we aim to investigate the expression and secretion of EDA+ FN from hepatocytes in nonalcoholic fatty liver disease (NAFLD) and the underlying mechanisms.Circulating levels of EDA+ FN were determined by ELISA in clinical samples.Western blotting and flow cytometry were performed on L02 and HepG2 cell lines to analyze whether the levels of EDA+ FN were associated with endoplasmic reticulum (ER) stress-related cell death.Circulating levels of EDA+ FN in NAFLD patients were significantly higher than those in control subjects,and positively related with severity of ultrasonographic steatosis score.In cultured hepatocytes,palmitate up-regulated the expression of EDA+ FN in a dose-dependent manner.Conversely,when the cells were pretreated with 4-phenylbutyrate,a specific inhibitor of ER stress,up-regulation of EDA+ FN could be abrogated.Moreover,silencing CHOP by shRNA enhanced the release of EDA+ FN from hepatocytes following palmitate treatment,which was involved in ER stress-related cell damage.These findings suggest that the up-regulated level of EDA+ FN is associated with liver damage in NAFLD,and ER stress-mediated cell damage contributes to the release of EDA+ FN from hepatocytes.

Objective To observe the effect of limited laminectomy combined with foraminal stenosis decompression on preventing C5nerve root palsy and improving neurological function. Methods From March,2014 to May,2016,69 patients with multi-segment cervical spondylotic myelopathy underwent surgical treatment in our hospital were included.Thirty-eight patients(group A)were treated with limited lami-nectomy combined with foraminal stenosis decompression and internal fixation,and 31 patients(group B)under-went routine laminectomy and internal fixation.The postoperative neurological recovery rate,cervical curvature index(CCI)and C5palsy rate were recorded and analyzed. Results No spinal cord and nerve injury occurred during the operation.The width of laminectomy was(16.8±2.1)mm in group A,and was significantly less than(21.7±2.5)mm in group B(t=8.849,P<0.001).There was no significant difference in operation time and intraoperative blood loss between two groups(t<0.439,P>0.05).The Japanese Orthopaedic Association (JOA) score increased continuously after surgery in both groups (F>42.996, P<0.05), and no significant difference was found between them at each time point(t<1.021,P>0.05).The cervical curva-ture index improved after surgery(F>86.379,P<0.05),and no significant difference was found between them at each time point(t<0.943,P>0.05).The spinal cord drift distance was(3.6±0.7)mm in group A,and ws signifi-cantly shorter than(2.5±0.5)mm in group B(t=7.602,P<0.001).There was no significantly difference in the neu-rological recovery rate between two groups(t=0.724,P=0.471).The C5palsy rate was lower in group A(5.2%,2/38)than in group B(22.5%,7/31)(χ2=4.514,P=0.034). Conclusion Limited laminectomy combined with foraminal stenosis decompression could relieve the spinal cord com-pression and restrict the excessive back drift, promote the neurological function recovery and reduce the inci-dence of C5palsy.

Objective　To study the effect of acute serious smoke inhalation injury to unilateral lung on contralateral lung and the therapeutic efficacy of massive bronchoscopic lavage with saline to the primary injured bronchoalveolus at early stage after smoke inhalation on mitigating the secondary damage of the other lung and so as to confirm the therapeutic validity of lung lavage to smoke inhalation injury. Methods　Fifteen mongrel dogs were insufflated with sawdust smoke into left lung and then randomized into 2 groups. The dogs in group A were maintained intravenous glucose saline transfusion and breathed air freely for 24 h after injury. Those in group B received a massive bronchoscopic lavage with 250 ml of saline to injured lung at 1 hour after injury and then were administrated similarly to those in group A. Gas exchange function of bilateral lungs in the process were observed and pathologic and phyiopathologic changes of the lung specimen were examined after the process. Results　While the total lavaging volume was kept to 250 ml, the liquid volume remained in left lung after each operating was from 90 ml to 140 ml. All animals had a significant decline in pulmonary function after smoke inhalation injury. PaO2 in group B decreased significantly soon after the lavage and then increased gradually to exceeding that in group A. The left lung showed serious edema similarly in two groups but in right lung the water content was less, dynamic and static compliance was more in group B than in group A. Conclusion　Bronchoalveolar massive lavage at early post-injury stage to injured lung after unilateral lung smoke inhalation injury can mitigate secondary damage of the non-injured lung. Lung lavage can decrease the intensity of secondary systemic inflammatory reaction and show a therapeutic validity to smoke inhalation injury.

Objective　To investigate the existence, intensit y and persisting time of biologic activity of bronchoalveolar lavage fluid (BALF ) collected from dogs with acute severe smoke inhalation injury in early post-i njury stage. Methods　BALF was collected 1 h after the est ablishment of acute severe sawdust smoke inhalation injury in 5 dogs, and the fl uid was perfused into the lungs of Wistar rats in the amount of 5 ml/kg (gro up C). Normal saline (group A) and BALF from normal dog (group B) were perfused into the lungs of rats and served as control. The respiratory rate, PaO2, lung water content and the expanding stability of lungs in all rats were determ ined at the time points of 4,12 and 24 h after the purfusion. Results　 Compared with the rats in group A and B, the rats in group C had higher mortality, wider range in RR, higher lung water content, PaO2 decreased obviou sly and lower lung expanding stability. The rats in control groups showed sl i ght mechanic obstruction in their airways in the course of experiment. Meanwhile ，the rats in group C showed higher oxidative activities and lower total anti-o xidative activities in lung tissues. Conclusion　It is certai n that the BALF collected from dogs with acute severe smoke inhalation injury in early post-injury stage could induce obvious injury in lung structure of norma l rats, showing certain mechanic obstruction in small airways. The injuring act ivity of the BALF can be alleviated ultimately 24 h after the perfusion of the B ALF.

To elucidate further sequence selectivity and nature of the binding of anticancer drugs to DNA, the interaction between anticancer drugs, which are minor groove ligands (distamycin A, DM and netropsin, NP) and intercalator (mitoxantrone, MT), and DNA were studied by electrospray ionization mass spectrometry. The 2 : 1 specific complex of DM and AT-rich DNA were observed principally, while only 1 : 1 specific complex of NP and AT-rich DNA were observed. MT specifically binds to GC-rich DNA. In addition, DM binds to DNA containing 5 A/T bases minor groove almost in a 2 : 1 mode and does not bind to DNA containing 3 A/T bases minor groove. NP binds most strongly to DNA containing 4 A/T bases minor groove. The 1 : 1 specific complex of MT and 6-mer DNA was also observed. The result of competitive binding experiment shows that DM binds more strongly to AT-rich DNA than NP does. These results provide bases for investigating the mechanism of interaction between the drugs and DNA and for improving the structure of target drug.
Antineoplastic Agents ; chemistry ; DNA ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; methods