Objective To improve the related substances analysis method of cobamamide.And to further research light degradation characteristics of cobamamide.Methods Determined the related substances of solid and aqueous solution of cobamamide after light degradation by high performance liquid chromatography(HPLC) conditions,which is Merck Hibar C18 column (4.6 mm ×250 mm, 5μm), 0.05 mol/L KH2PO4 solution:acetonitrile (90:10) -80% acetonitrile as mobile by gradient elution and detection wavelength 260 nm.And compared their light stability.The main three kinds of light degradation impurities were determinate from LC-MS.Results Gradient elution made the light degradation impurities separate better.The results of precision and reproducibility tests increased to RSD =0.2% (n=6) and RSD =8% (n=5) from RSD =3.8% (n =6) and RSD=38%(n=5). Cobamamide solution was very sensitive to light, the preparation should be strict dark operation.Two of the light degradation impurities were adenosine and hydroxycobalamin, with the relative response factor 2.5 and 0.7.Conclusion New method is specific, durable and reproducible, which can be used for quality control of cobamamide.

Animals ; Enterovirus A, Human ; genetics ; Enterovirus Infections ; virology ; Humans ; Reverse Genetics ; methods

BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) are conveniently cultured and separated in vitro because theirimmunogenicity is low. Therefore, BMSCs are suitable for cell transplantation. Research has shown that BMSCs are potential to repair neurological defect. OBJECTIVE: To determine whether in vitro cultured BMSCs can be transplanted to repair peripheral nerve injury or not, and to investigate its mechanisms. DESIGN, TIME AND SETTING: Randomized controlled animal study This study was performed in Department of Toxicology, Public Health College of Jilin University from March 2006 to March 2007.MATERIALS: Fifty healthy female Wistar rats aging 2 months and six 1-week-old female Wistar rats were used for extraction of BMSCs. Rabbit-anti-nerve growth factor (NGF) monoclonal antibody was provided by Santa Cruz Company. METHODS:BMSCs were separated and cultured with adherent method. In the 3rd generation, BMSCs were preiabeled with bromodeoxyuridine (BrdU) 48 hours before transplantation. Fifty healthy Wistar rats were selected to prepare sciatic nerve crush injury models with clamping method.Subsequently, rats were randomly divided into transplantation group and control group, with 25 rats in each group. Rats in the transplantation group underwent transplantation of BrdU-labeied BMSCs at nerve injured sites; while, the same volume DMEM was injected into rats in the control group. MAIN OUTCOME MEASURES: Injured nerve in the transplantation group suffered from anti-BrdU staining 1, 2, 4, and 6 weeks after surgery. Distal injured nerve in both groups suffered from NGF immunohistochemical staining 1, 2, 4, and 6 weeks after surgery. Image analysis system was adopted to analyze integrated absorbance of positive expression. Gait analysis was performed every week after surgery to measure sciatic nerve function index, and it was also adopted to measure regenerated nerve conduction velocity 6 weeks after surgery. Subsequently, amount and inner diameter of medullated nerve fibers were calculated after luxol fast blue staining, while wet weight of experimental-lateral gastrocnemius muscle and cross section area of muscle fiber were measured at the same time. RESULTS: Fifty rats were included in the final analysis. BrdU-labeled positive cells could be found at injured nerve in the transplantation group 1, 2, and 4 weeks after surgery. Integrated absorbance of NGF protein expression in the transplantation group was significantly higher than that in the control group 1 and 2 weeks after surgery (P < 0.01), but there were no significant differences between the two groups 4 and 6 weeks after surgery (P > 0.05). Sciatic nerve function index in the transplantation group superiorly recovered to that in the control group 3-6 weeks after surgery. Furthermore, 6 weeks after surgery, nerve conduction velocity, amount and diameter of medullated nerve fibers, wet weight and cross section area of gastrocnemius muscle in the transplantation group were significantly higher than those in the control group (P < 0.05-0.01). CONCLUSION: BMSCs can be transplantated into injuried nerve tissue, and promote the recovery of nerve function in the micro-enviroment, improve NGF expression in an early phase may be one of its mechanisms.

PurposeAs a main method to detect thyroid nodules, ultrasonography seems to have a rather low accuracy in differentiating benign and malignant ones. The present study aims to explore the diagnostic value of thyroid imaging reporting and data system (TI-RADS) in identifying benign and malignant thyroid nodules.Materials and Methods A total of 168 patients with thyroid nodules conifrmed pathologically (with 251 suspicious nodules) underwent ultrasonography and were further grouped into category 3-5 according to TI-RADS classiifcation standard. The results were retrospectively compared with histopathological ifndings. The sensitivity, speciifcity, accuracy, positive predictive value and negative predictive value of TI-RADS classiifcation in the diagnosis of benign and malignant thyroid nodules were calculated, and the differences in ultrasonic features between benign and malignant thyroid nodules were also compared.Results The surgical and pathological findings showed that 94 nodules were benign and 157 were sinister; TI-RADS regarded 93 nodules were benign and 185 were malignant. The difference of the two means of identification was significant (χ2=149.6,P<0.01). The sensitivity, speciifcity, accuracy, positive predictive value and negative predictive value of TI-RADS were 91.2% (144/157), 85.1% (80/94), 89.3% (224/251), 91.2% (144/158) and 86.0% (80/93), respectively. The ultrasonic manifestations of benign and malignant nodules were signiifcantly different with regard to aspect ratio, echogenicity, shape, calciifcation inside the nodule (χ2=8.7-121.4,P<0.01).Conclusion TI-RADS classiifcation standards have a high accuracy in the differentiation of benign and malignant thyroid nodules as a means of ultrasound examination, and thus can serve as an important guiding method in clinical diagnosis and treatment.

While the medicine pattern of biomedicine turn to biological-psychology-society, the medical trouble communication becomes more and more important in the medical service. Good medical trouble communication ability is the essential condition of doctor. As oral cavity clinicians, only by gasping the principle of communication can we appropriately utilize some skills of communication exchange,establish the good medical trouble relations with the patient and achieve the good treatment result finally.

Aim Drug resistance is one of the major hinders on cancer treatments. α-enolase ( eno1 ) was closely related to the generation and development of drug resistance. This article aims to study the effect of eno1 on cell growth and drug resistance in human chro-nic myeloid leukemia cell line K562/A02 . Methods We screened three eno1 stable silencing cells K562/A02-sheno1 and its control cells K562/A02-shcon. Cell count assay was performed to test cell growth, MTT assay was used to test cell proliferation, flow cytometry was used to test the intra-cellular Rho123 content, the expression of genes were tested by real-time PCR assay and western blot assay on mRNA level and protein level, respectively. Results eno1 was o-ver-expressed in K562/A02 cells and its expression was increased by ( 2. 85 ± 0. 56 ) times and ( 1. 43 ± 0. 05 ) times on mRNA level and protein level com-pared to K562 cells. However, there was no difference in cell growth rate between K562/A02 cells and K562 cells. K562/A02-sheno1 cells showed lower cell growth rate and higher drug sensitivity to anti-cancer drugs taxol and doxorubicin. Moreover the Rho123 content was increased in K562/A02-sheno1 cells. The expression of MDR1 decreased in both mRNA level and protein level in K562/A02-sheno1 cells. Conclusion eno1 silencing could suppress cell growth, reverse drug resistance and increase its drug sensitivity in K562/A02 cells, and the mechanism was associated with the MDR1 gene.

Objective To investigate that the relationship of serum concentration of copeptin ,procalcitonin(PCT )and early diagnosis ,prognosis in patients with cerebral hemorrhage‐associated pulmonary infection .Methods One hundred and twenty pa‐tients with acute cerebral hemorrhage ,acute cerebral hemorrhage‐associated pulmonary infection and 60 healthy individuals (the control group) admitted in our hospital from January 2012 to December 2013 were selected and detected for serum copeptin and procalcitonin .The differences of serum copeptin ,procalcitonin levels were compared in controls ,in patients with cerebral hemor‐rhage and cerebral hemorrhage‐associated pulmonary infection and their correlation was analyzed .Results The levels of serum copeptin in the cerebral hemorrhage‐associated pulmonary infection were significantly higher than that in control group and the cer‐ebral hemorrhage (P<0 .05) .The levels of serum procalcitonin in control group and the cerebral hemorrhage were significantly lower than that in the cerebral hemorrhage‐associated pulmonary infection ,the levels of serum C‐reactive protein ,copeptin ,procalci‐tonin and the APACHEⅡ scores of the patients with survival group were significantly lower than those with non‐survival group (P<0 .05) .Conclusion The levels of serum copeptin and procalcitonin are correlated intimately with cerebral hemorrhage‐associat‐ed pulmonary infection .They are important clinical parameters to reflect the early diagnosis and prognosis of cerebral hemorrhage‐associated pulmonary infection .

Objective To explore the effects of herpes simplex virus 2 (HSV-2) latency-associated transcript open reading frame 3 (LAT ORF3) gene on Vero cells against cisplatin-induced apoptosis.Methods Recombinant plasmid enhanced green fluorescent protein-open reading frame 3 (named pEGFP-ORF3) was constructed and transfected into Vero cells; then,reverse transcription (RT)-PCR was performed to detect the expression of the target gene.Cisplatin of 3 mg/L was selected to induce the apoptosis in Vero cells.Cultured Vero cells were transfected with empty plasmid and induced by cisplatin (pEGFP-C2 group),transfected with recombinant plasmid pEGFP-ORF3 and induced by cisplatin (pEGFP-ORF3 group),only induced by cisplatin (cisplatin-induced control group),or remained untreated (normal control group).Subsequently,fluorescence microscopy was conducted to observe apoptotic bodies,Giemsa stain to observe the morphology of cell nuclei,methyl thiazolyl tetrazolium (MTT) assay to evaluate cell proliferation,and flow cytometry to assess cell apoptosis.Data were assessed by using SPSS 13.0 software,and statistical analysis was carried out by one-way ANOVA and t test.Results HSV-2 333 LAT ORF3 gene was successfully cloned.The eukaryotic expression plasmid for LAT ORF3 was constructed,and the expression of LAT ORF3 gene in Vero cells was confirmed by RT-PCR.Giemsa stain showed blue-staining nuclei and pale cytoplasm in recombinant plasmid-transfected and cisplatin-induced Vero cells with a normal shape.The value of cell proliferation (absorbance at 490 nm) by MTT assay was 2.56 ± 0.21 in pEGFP-ORF3 group,similar to that in the normal control group (2.66 ± 0.13,P ＞ 0.05),but significantly higher than cisplatin-induced control group (1.65 ± 0.11,P ＜ 0.05) and pEGFP-C2 group (1.56 ± 0.18,P ＜ 0.05).As far as the apoptosis rate was concerned,no significant difference was observed between pEGFP-ORF3 group and normal control group (4.03％ ± 1.04％ vs.2.13％ ± 0.09％,P ＞ 0.05),but pEGFP-ORF3 group was statistically lower than pEGFP-C2 group (19.45％ ± 2.05％,P ＜ 0.05).Conclusion The transfected HSV-2 LAT ORF3 gene could protect Vero cells from cisplatin-induced apoptosis.