Characteristics and measurement principles of reference methods in clinical biochemistry were described.Implementation of reference systems is one of the most effective approaches to improve the accuracy and comparability of clinical laboratory test results.Reference methods are the key components of reference systems.Reference methods should have measurement uncertainties that meet the requirements of the intended use,and thus should be based on reliable measurement principles.For the well-defined biochemistry analytes,reference methods have been almost all based on instrumental analysis.Isotope dilution mass spectrometry (ID/MS) is considered most reliable and has been the major analytical principle of the reference methods.ID/MS analysis is accurate but expensive.Use of other validated instrumental analyses as reference measurement principles would be justified.

Proteolytic degradation has been a severe problem when Pichia pastoris is employed to express recombinant proteins.One alternative method to circumvent this problem is to construct protease gene disruptant.However,the main study of gene disruption is focused on nonrecombinant Pichia pastoris rather than recombinant strain.In our study,we established two different methods to directly disrupt PRC1 and KEX1 gene in recombinant Pichia pastoris.On the basis of this,we further discussed and compared the application and advantages of both methods.

Objective To evaluate high resolution manometry in the diagnosis of hiatal hernia.Methods Clinical data were reviewed on 20 patients suffering from gastroesophageal reflux who had laparoscopic Toupet fundoplication for preoperative tentative diagnosis of hiatal hernia.Preoperative diagnosis of hiatal hernia was made collectively by endoscopy,X-ray examination,24 hour esophageal pH monitoring and high resolution manometry before surgery.Results Preoperative diagnosis of hiatal hernia was made in 3 patients by X-ray examination,in 9 patients by high resolution manometry.11 patients were finally diagnosed with hiatat hernia intraoperatively.X ray was consistent with intraoperative diagnosis in 27％ cases.Intraoperative and endoscopic diagnoses were 55％.High resolution manometry and intraoperative diagnoses were consistent in 82％.Lower esophageal sphincter length was (1.92 ± 0.38) cm in hiatal hernia group and (2.10 ± 0.92) cm in non-hiatal hernia group (t =0.60,P ＞ 0.05),lower esophageal sphincter pressure (respiratory min) was (0.64 ±0.55) kPa in hiatal hernia group and (1.31 ± 1.07) kPa in nonhiatal hernia group(t =1.80,P ＞ 0.05),and lower esophageal sphincter pressure (respiratory mean) was (1.43 ±0.92) kPa in hiatal hernia group and (2.57 ± 1.33) kPa in non-hiatal hernia group(t =2.26,P ＜0.05).The reflux parameters,including the percent total time pH ＜ 4,and DeMeester score,were significantly greater in hiatal hernia group than in non-hiatal hernia group (all P ＜ 0.05).Conclusions Hiatal hernia patients are with poor esophageal antireflux competency and severe reflux.High resolution manometry is more valuable in the diagnosis of hiatal hernia than endoscopy or X-ray examination.

Objective To discuss the feasibility and curative effect of broncho-alveolar lavage(BAL)through bronchofiberscope in the treatment of severe toxic pulmonary edema caused by irritative gas.Methods 16 cases se- vere toxic pulmonary edema caused by irritative gas were performed BAL through bronchofiberseope.The index of oxygen in arterial blood,clinical and radiological changes before,during and 2 hours after BAL were observed.Results 2 hours after BAL through bronchofiberscope,the partial pressure of oxygen in arterial blood(PaO2)obviously in- creased,the partial pressure of carbon dioxide in arterial blood(PaCO_2)did not change much.PaO_2 and PaCO_2 had no obvious change before and during BAL.The shadow area in the X-ray film of chest obviously decreased 24 hours af- ter BAL.In all 16 cases,13 cases were cured,1 case got improvement,and 2 cases died.The curative rate was 81%. Conclusion BAL through bronchofiberscope could clear the noxious substance in airway and improve the ventilation function.It was safe and had confirmed curative effect.

Objective To choose proper proton magnetic resonance spectroscopy(~1XH-MRS) parameters to fit for practical femoral marrow cavity and to produce short-timed,well-repeated and excellent ~1H-MRS images.Methods The tentative study of ~1H-MRS on the normal femoral bone marrow in 26 volunteers was performed with a 1.5 T MR after the informed consent.The single-voxel spectroscopy and stimulated echo acquisition mode were used for ~1H-MRS collection.~1H-MRS parameters for 12 volunteers were 128 acquisitions,1 cm?1 cm?1 cm volume of interest(VOI)size and repeatedly 2—3 times within the same location.~1H-MRS parameters for another:14 volunteers were different numbers of acquisition (128 and 256 times,respectively)and different VOI sizes(2 cm?2 cm?2 cm and 1 cm?1 cm?1 cm, respectively).Results For ~1H-MRS with 1 cm?1 cm?1 cm size of VOI and 128 times of acquisition with the full width haft max of water≤8—12 Hz,the base-line was steady and the signal-noise ratio was high up to 11.31.~1H-MRS was different in the different femoral locations showing the maximum peak sites at near 0.90 ppm(?10~(-6))or 1.65 ppm,but~1H-MRS within the same location was always same or similar with different VOI sizes(1 cm?1 cm?1 cm or 2 cm?2 cm?2 cm)or different numbers of acquisition(128 or 256 times).~1H-MRS acquisition time was not related with the size of VOI but with the numbers of acquisition.128 and 256 times of acquisition cost 199 s and 391 s,respectively.Conclusion With the technique of small size of VOI(1 cm?1 cm?1 cm)and decreased numbers of acquisition(128 times),it is propable to get well-repeated and excellent ~1H-MRS within less time.It is also more practical for clinics to achieve ~1H-MRS of the femoral marrow with the proper technique.

?AIM: To evaluate the effects on recovering of corneal wound and postoperative discomfort of different methods for primary pterygium.?METHODS: Forty-seven cases ( 60 eyes ) of primary pterygium were excised under microscope with limbal epithelial transplantation, with sharp dissection ( 24 cases, 30 eyes, Group A) and blunt dissection (23 cases, 30 eyes, Group B).All cases were followed up for 1d to 1mo.?RESULTS: The recovering of corneal wound was better in Group B on 1st day and 3rd day after surgery.Pain, photophobia and tears, foreign body sensation were more serious in group A on 1st day after surgery with a statistically significant difference (P=0.005,0.015,0.012). Pain, photophobia and tears, foreign body sensation were more serious in Group A on 3rd day after surgery with a statistically significant difference ( P=0.019,0.018, 0.015).There was no statistically significant difference on 1wk and 1mo after surgery (P>0.05).? CONCLUSION: Compared with sharp dissection, primary pterygium excised with blunt dissection can significantly improve recovering of corneal wound and postoperative discomfort.

Objective: To observe the level of pro-opiomelanocortin (POMC) mRNA expression in the hypothalamic arcuate nucleus in young and senile rats. Methods: POMC mRNA was determined with in situ hybridization, the hybridization signals and areas of POMC mRNA-positive cells were measured with computer image analysis system. Results: Compared with young rats, average gray values of POMC mRNA-positive neurons in the hypothalalmic arcuate nucleus and near region in the senile rats decreased, but the numbers of POMC mRNA-positive neurons did not change significantly. Conclusion: The decrease of POMC mRNA expression level in individual neurons in hypothalamic arcuate nucleus of senile rats is one of the causes that decrease the expression of peptides-derived from POMC gene.

OBJECTIVETo investigate the regulatory effects of nerve growth factor (NGF) on basal and capsaicin-induced release of neuropeptide substance P (SP) in primary cultured embryonic rat dorsal root ganglion (DRG) neurons.

METHODSDRGs were dissected from 15-day-old embryonic Wistar rats. DRG neurons were dissociated and cultured, and then exposed to different concentrations of NGF (10 ng/mL, 30 ng/mL, or 100 ng/mL) for 72 h. The neurons cultured in media without NGF served as control. RT-PCR were used for detecting the mRNAs of SP and vanilloid receptor 1 (VR1) in the DRG neurons. The SP basal and capsaicin (100 nmol/L)-induced release in the culture were measured by radioimmunoassay (RIA).

RESULTSSP mRNA and VR1 mRNA expression increased in primary cultured DRG neurons in a dose-dependent manner of NGF. Both basal release and capsaicin-evoked release of SP increased in NGF-treated DRG neurons compared with in control group. The capsaicin-evoked release of SP also increased in a dose-dependent manner of NGF.

CONCLUSIONNGF may promote both basal release and capsaicin-evoked release of SP. NGF might increase the sensitivity of nociceptors by increasing the SP mRNA or VR1 mRNA.

Analgesics, Non-Narcotic ; pharmacology ; Animals ; Capsaicin ; pharmacology ; Cells, Cultured ; Dose-Response Relationship, Drug ; Embryo, Mammalian ; Ganglia, Spinal ; cytology ; Gene Expression Regulation ; drug effects ; Nerve Growth Factor ; pharmacology ; Neurons ; drug effects ; RNA, Messenger ; metabolism ; Radioimmunoassay ; methods ; Rats ; Rats, Wistar ; Substance P ; genetics ; metabolism

Objective:To investigate the expression of Fas、FasL and the apoptosis of liver cancer cell line HepG2 transfected with LIGHT and IFN-? gene mediated by Cationic liposome.Methods:HepG2 cells were divided into two groups(the solo transfection of LIGHT gene and the combined transfection of LIGHT and IFN-? genes) and the control groups(no transfection).HepG2 cells were cellected at 12h,24h and 48h after transfection.The apoptosis of HepG2 cells and the expression of Fas and FasL of the HepG2 cells were investigated with flow cytometry.Results:After transfection,the apoptosis of HepG2 cells increased,and the apoptosis of combined transfection group was higher than the solo transfection of LIGHT(P

Forty batches of Lonicerae Japonica Fse i collected extensively and prepared as the test solution. Their chromatographic fingerprints and anti-influenza virus IC50 value (half maximal inhibitory concentration) were determined respectively. Then Unscrambler software was used, and spectrum-efficient correlation analysis was done for chromatographic fingerprints data and IC50 data by partial least squares regression method, to establish spectrum-efficient correlation model for anti-influenza virus of Lonicerae Japonicae Flos. Then the other 10 batches of Lonicerae Japonicae Flos were used to verify the model and explore the adaptability of this spectrum-efficient correlation model based on partial least squares regression method. The mathematical model obtained R2 of 0.969489 and RM-SEC of 0.070691 for calibration set; R2 of 0.959042 and RMSECV of 0.084005 for cross validation set. The verification experiment results showed that the relative error between the predicted values and measured values was within 10% in all 10 hatches, and within 5% in 80% of them. The results showed that the established spectrum-efficient correlation model could be used to evaluate the biological activity of anti-influenza virus of Lonicerae Japonicae Flos by determining its HPLC fingerprints.
Antiviral Agents ; analysis ; pharmacology ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; pharmacology ; Flowers ; chemistry ; Least-Squares Analysis ; Lonicera ; chemistry ; Molecular Structure ; Orthomyxoviridae ; drug effects