Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) is a widely used technique in the quantitative analysis of small molecules, part of peptides and proteins in biological matrix. When compared with LC, the LC-MS/MS posesses many advantages, such as high sample throughput, high sensitivity and resolutions, simple method development and simultaneous quantification of multiple components. At the stage of method development, the main practices for analysts are to validate analytical methods according to the regulated requirements. However, due to the quantitative mechanisms of LC-MS/MS, even though the used method is fully validated, analytical inaccuracy problem, which is the so called bioanalytical risk (pitfalls), may still exist, if we do not pay attentions to some details. In the present paper, the factors which may cause inaccuracy in quantitative analysis are summarized, especially those could not be found during routine method validation. Those factors include back transformation of unstable drug metabolites to analyte in the handling of biosamples, matrix effects, in-source conversion, and ion interference caused by ions with similar or same mass-to-charge ratio, etc. In addition to QA/QC oversight and strict method validation, the preventing strategies may contain the follows: selecting biosample processing method with high proficiency in eliminating matrix interferences, enhancing chromatographic separation, selection of suitable precursor-product ions and validation with incurred samples, etc.

Objective　According to Kaschin-Beck Disea se monitory standardization that had been adjusted by our country,we monitored the state of Kaschin-Beck Disease in Gansu province.Methods　So as to understand change of illness,we took methods of epidemiological investigation,clinical examination and X-ray diagnosis.Results　It is not detected in the clinical that patient suffered from more than I of KBD among 7～12 years old in Qingyang monitory netw ork.X-ray detectable rate is 3%,but 12 cases patients were showed in Zhangjiach uan.X-ray detectable rate is 22.22%.Conclusions　Illness was showed steady state and was con trolled in Qingyang region,but illness recurred clearly in Zhangjiachuan region.

Objective To study the relationship of surgical procedures with clinical effect in herniat-ed lumbar disc in order to improve operative methods and obtain a better outcome.Methods Retrospec-tive analysis was carried out in7235patients with herniated lumbar disc,who had been operated by removal of nucleus pulposus using small incision and fenestration in our hospital since1983,313cases of whom re-ceived second operation because of postoperative complications.There were187males and126females aging from27to62years(mean,45.8years),the incidence of morbidity was4.32%.During the same period,552patients,who had less approved operative result primarily treated in other hospitals,were admitted to our hospital.There were317males and235females aging from31to64years(mean,46.0years).Results The postoperative complications could be divided into short term(within1month after operation)and mid-dle to long term(more than1month after operation)groups.The total446cases with short term compli-cation were133cases primarily treated in our hospital and313cases in other hospital,including of re-herni a-tion or incomplete nuclear extirpation in77treated with second operation,discitis in106treated with con-servative therapy for 67and second operative management for 39,canal hematoma compression in76treated surgi cally,multi-level herniation and leakage in21and mislocalization in59re-operated,injury of nerve root or cauda equina in85treated with nerve exploration,release and anastomosis,residual materials in canal in17with removal of foreign body,venous thrombus in2,and arachnoiditis in3.The total424cases with middle to long term complications were185cases primarily treated in our hospital and239cases in other hospitals,including of nerve root adhesion in159,recurred disc herniation in122,segmental instability in81,and ia-trogenic spinal stenosis in62.Conclusion Many factors may influence the outcome of herniated lum bar disc which can be abolished by sufficient preparation,careful operation and proper managememt.

MTT Colorimetric method is usually applied for measuring the living animal cell number. By changing the reaction temperature and the reaction time as well as the colorimetric wavelength, the improved MTT colorimetric method was established to count the living bacterial cell number. This new method was used to measure the living cell concentration in the process for culturing bacteria PBW1. The results measured by the improved MIT colorimetric method and dilute plate method are similar. Compared with other methods including the dilute plate method, the improved MTT colorimetric method has many advantages such as accuracy, quickness.

The crude toxin was extracted from hypha and culture solution of Phyllosticta commelimecola through three different polarity solvent: benzinum, puncificatum ethyl acetate and chloroform. The result indicated that the toxin secreted by Phyllosticta commelimecola not only was in hypha but also in culture solution and the extracting effect of ethyl acetate was the best. The soybean median and PSK media can be respectively used as solid and liquid culture media to produce toxin and grow mycelium. The optimal cultural conditions for producing toxin were temperature 32℃,cultured period 14d, cultured ways shaking of 150r/min.

BACKGROUND: Fracture of the proximal femur is common in the elderly. Biological artificial femoral head replacement is one of the main methods of surgical treatment, but in the patients with chronic renal disease, the surgical efficacy and safety are variously reported. OBJECTIVE: To analyze the middle- and short-term effects of biological artificial femoral head replacement in elderly patients with proximal femoral fracture of chronic renal disease. METHODS: Biological artificial femoral head replacement was used to treat 53 patients with proximal femoral fractures (aged 75 years and over). They were divided into two groups according to renal function: chronic renal disease group (n=25) and non-chronic renal disease group (n=28). Operation time, intraoperative blood loss, intraoperative blood transfusion, postoperative drainage volume of 24 hours, and partial and complete weight-bearing time, hospitalization time, hospitalization expenses and complications were compared between the two groups. Harris hip joint function score, dual energy X-ray bone density, prosthesis loosening, periprosthetic osteolysis and implant survival rate were compared in the two groups during follow-up. RESULTS AND CONCLUSION: (1) In the non-chronic renal disease group, one patient died of respiratory failure at 1 month after replacement, and one case was refurbished at 3 months after replacement. In the chronic renal disease group, one patient died of cardiopulmonary failure at 3 months after replacement. Other patients in both groups were followed up for 1-8 years. The repair efficacy was satisfactory. (2) The excellent and good rate of hip function Harris score was 83% and 85% at 6 months after operation in chronic renal disease group and non-chronic renal disease group, respectively (P > 0.05). (3) No significant differences in operation time, blood loss, the volume of drainage 24 hours after the operation, the partial and complete weight-bearing time and dual energy X-ray bone density were determined between the two groups. The blood transfusion was larger; hospitalization time was longer; hospitalization expenses were more in the chronic renal disease group compared with the non-chronic renal disease group. (4) Patients in the two groups experienced different degrees of osteolysis, but no obvious prosthesis loosening was found. One patient in the non-chronic renal disease group underwent revision because of repeated dislocation of the prosthesis. The survival rates of the prosthesis in the chronic renal disease group and non-chronic renal disease group were 100% and 96% respectively (P > 0.05). (5) For elderly patients with proximal femoral fracture of chronic renal disease, the biological artificial femoral head replacement obtained good repair efficacy, and the function of the limbs was improved obviously with careful preoperative preparation.

AlM: To describe the expression of hypoxia-inducible factor-1α ( HlF-1α) and erythropoietin ( EPO ) in rats' corneal and evaluate its potential effect on corneal neovascularization ( CNV) growth.
METHODS:The young SD rats (3mo) was chosen and randomly divided into 2 groups, which were experimental group and normal control group. CNV model was established by alkali burn, and the length and area of CNV was observed everyday after operation by slit lamp. After that, the expression of HlF-1α and EPO was measured by SABC and RT-PCR methods at 1, 3, 5, 7, and 14d after alkali burn. The data was analyzed by SPSS 20. 0.
RESULTS:The area of CNV was increasing at 1, 3, 5, 7, and 14d after alkali burn, and the peak point appear at 7d. The growth speed was decreased after 14d. SABC method told us that no HlF-1αand very tiny amount EPO was detected at normal rats' corneal. The expression of the two factors increased at 1d after alkali burn in corneal epithelium and endoderm. The results of RT - PCR showed that a few amounts of HlF-1α and EPO mRNA were detected at normal group. The expression of the two factors was increased at 3d after alkali burn, and the peak value was found at 7d, however, it was decreased at 14d. Statistical difference was found at different time (P<0. 05).
CONCLUSlON: HlF- 1α and EPO is closely related to CNV.

AlM: To demonstrate the effect of comprehensive treatments for meibomian gland dysfunctional dry eye.
METHODS:Totally, 86 cases (172 eyes) in our hospital, who suffered from meibomian gland dysfunctional dry eye, were measured by BUT, CFS, the machine of infrared meibomian gland function and testing meibomian gland after extrusion. The scores of meibomian gland function missing and secretions were observed, which as a diagnosis, and assessed the effectiveness of treatment, after took comprehensive treatments in different stages ( comprehensive treatments including cleaning, hot covering, massaging, TobraDex, Pranoprofen and Lubricant, etc. ) .
RESULTS:All patients were treated and observed for 3wk, in which 46 cases (92 eyes) of cured, 29 cases (58 eyes) effective, 11 cases (22 eyes) ineffective, the total effective rate was 87. 2%. After 6wk, 68 cases (136 eyes) of cured, 13 cases (26 eyes) effective, 5 cases (10 eyes) ineffective, the total effective rate was 94. 2%, not appeared adverse ocular and systemic reactions in the process of treatment.
CONCLUSlON: Taken comprehensive treatments for meibomian gland dysfunctional dry eye could effectively shorten the course of the disease, quickly remove meibomian gland tube obstruction, restore the meibomian gland secretion and stable the lipid layer, and could reduce the discomfort of patients with dry eye positively and effectively.

Objective To investigate whether IL-17 could stimulate the vascular endothelial growth factor (VEGF) production on HaCaT cells alone. We also investigated whether shikonin could inhibited the proinflamation effects of interleukin-17(IL-17) acting on HaCaT cells. MethodsWe examined the expression of VEGF by double antibody sandwich enzyme-linked immunosorbent assay ( ELISA ) and realtime polymerase chain reaction(RT-PCR) in HaCaT cells and the cell supernatant. The viability of HaCaT cells in the drug group was detected by the Cell Counting Kit-8 (CCK-8). ResultsThe expression of VEGF in different time IL-17-stimulated groups on HaCaT cells and the cell supernatant were higher than the control group( P＜0.001 ). The expression of VEGF in different drug treatment groups on HaCaT cells and the cell supematant were lower than the stimulated group by IL-17 ( P＜0. 001 ). The cell viability of different drug treatment groups have no significant difference( P＞0.05 ). ConclusionWe show that IL-17 specifically and time-dependently augmented and induced VEGF expression on HaCaT cells and the cell supernatantThen shikonin markedly inhibited the increase tengency of IL-17 effection on HaCaT cells and the cell supematant level.