OBJECTIVE: To determinate triptolide and wilforlide A in biological samples by liquid chromatography-tandem mass spectrometry (LC-MS/MS) method and to verify the method. METHODS: After 0.4 mL blood, urine or 0.4 g hepatic tissues with internal standard were extracted by ethyl acetate, they were separated on a Allure PFP Propyl (100 mm x 2.1 mm, 5 µm) with a mobile phase of methanol-20 mmol/L ammonium acetate using gradient elution. For mass spectrometric detection, electrospray ionization (ESI⁺) in positive mode was elected and the data was collected using multiple-reaction monitoring (MRM). RESULTS: The linearity was good (r > 0.995 0) and the limit of detection was 2 ng/mL or 2 ng/g for triptolide and wilforlide A. The recovery was 61.08%-102.98%. The intra-day and inter-day precision was less than 12.58% for each biological sample, and the accuracy was 90.61%-105.80%. CONCLUSION This method is simple, convenient and good selective, and could be applied to analysis of triptolide and wilforlide A in different biological samples. And the method may provide technical support for forensic medicine identification, clinical diagnosis and treatment of tripterygium wilfordii Hook. f. poisoning.
Chromatography, High Pressure Liquid ; Diterpenes/urine* ; Epoxy Compounds/urine* ; Humans ; Oleanolic Acid/urine* ; Phenanthrenes/urine* ; Sensitivity and Specificity ; Tandem Mass Spectrometry

AlM:To investigate the effect of peripheral vision control technology for delaying the development of juvenile myopia.METHODS:A total of ninty-nine cases of 12 ~18 year-old myopic patients were randomly divided into two groups. The experimental group was peripheral vision control technology group whose members wore the special lenses which can help correct the hypermetropic defocus of peripheral retina. The other was control group whose members wore ordinary monofocal lenses. All the subjects needed to accept re-examination every 3mo and be recorded the data of dioptre, corneal curvature and axial length.RESULTS: After 18mo, the dioptre and axial length of two groups had increased in varying degree. But the data's different quantity of the corneal curvature in each group had no statistical significance ( P > 0. 05 ). Themyopia deepen quantity in experimental group was-0. 65±0. 65D and its axial growth was 0. 23±0. 22mm, and the myopia deepen quantity in control group was -1. 17 ± 0. 50D and its axial growth was 0. 41 ± 0. 17mm. Under the circumstance of valid data, the increment of myopia and axial length in experimental group was lower than that in control group (P<0. 05).CONCLUSlON: Peripheral vision control technology can delay the development of juvenile myopia effectively.

OBJECTIVE: To develop a liquid chromatography-tandem mass spectrometry (LC-MS/MS) screening method for 45 poisonous alkaloids in blood. METHODS: Identification was based on the compound's retention time and two precursor-to-production transitions. The method involved a liquid-liquid extraction (LLE) followed by LC-MS/MS with multiple-reaction monitoring (MRM). When 1 mL of blood was extracted with diethyl ether at pH = 9.2 with SKF525A as the internal standard, the target compounds were analyzed with LC-MS/MS in the positive ionization mode. RESULTS: The target alkaloids had good linearity (r>0.995 1), both the intra-day precision and inter-day precision being less than 14.77%. The limits of detection ranged from 0.05 to 25 ng/mL in blood. CONCLUSION The method is selective and sensitive in detecting poisonous alkaloids with a total running time of 12 minutes; therefore it was successfully applied to some actual cases of suspected alkaloids poisoning.
Alkaloids/chemistry* ; Chromatography, Liquid/methods* ; Forensic Medicine ; Humans ; Liquid-Liquid Extraction ; Mass Spectrometry ; Sensitivity and Specificity ; Tandem Mass Spectrometry/methods*

Objective To observe relationship between the phlegm and adhesion molecule and further explore the functions of the phlegm in the metastatic potential of the tumors. Methods Examin the expression difference of ICAM-1,E-cad,MMP-9 of gastric cancer by enzyme linked immunosorbent assay between the phlegm-stamp type and non-phlegm-stamp type,then reducing phlegm by chinese medicine treatment and comparing theirs expression variation.Results The expressions of ICAM-1,E-cad, MMP-9 were obviouslv different between the phlegm group and non-phlegm group[(403.6±99.7)μg/L,(9.08±1.69)mg/L,(465.0±96.64)μg/L &(319.9±81.4)μg/L,(7.56±1.15)mg/L,(228.1±43.79)μg/L].After using xiao-tan-san-jie recipes,the expressions of ICAM-1 and E-cad of the phlegm group were obviously lower than before(P<0.05);the expression of MMP-9 was also falling down,but the difference was not significative.Conclusion There is a correlation between the phlegm and adhesion molecule, which could take action in tumor metastams by affecting adhesion molecule expression.

AIM:To investigate how to design personalized cataract surgery programs to achieve surgical correction of preoperative corneal astigmatism with surgical astigmatism under the guidance of corneal topography, improve postoperative visual quality and reduce the cost of treatment.
METHODS: Totally 202 cases ( 226 eyes ) cataract patients were divided into randomized treatment group and individualized treatment group. According to the method and location of the incision, randomized treatment group were divided into 8 groups. Surgical astigmatism after different incision were calculated with the use of preoperative and postoperative corneal astigmatism through vector analysis method. Individualized treatment groups were designed personably for surgical method with reference of every surgically induced astigmatism, the surgical method chooses the type of surgical incision based on close link between preoperative corneal astigmatism and surgically induced astigmatism, and the incision was located in the steep meridian. The postoperative corneal astigmatism of individualized treatment group was observed.
RESULTS: Postoperative corneal astigmatism of individualized treatment group were lower than that of 3.0mm clear corneal tunnel incision in the randomized treatment group, there were statistically significance difference, while with 3. 0mm sclera tunnel incision group there were no statistically significance difference. After 55. 8% of patients with the use of individualized surgical plan could undergo the operation of extracapsular cataract extraction with relatively low cost and rigid intraocular lens implantation, the per capita cost of treatment could be reduced.
CONCLUSION: Personalized cataract surgery programs are designed to achieve surgical correction of preoperative corneal astigmatism under the use of corneal topography, improve postoperative visual quality and reduce the cost of treatment.

OBJECTIVETo invesitigate the clinical effects of percutaneous closed reduction and cannulated screw internal fixation combined with external fixation in the treatment of unstable pelvic fractures.

METHODSFrom April 2006 to May 2009,29 patients with pelvic fractures of rotatory instability were treated with closed reduction and screw internal fixation combined with external fixation. There were 19 males and 10 females with an average age of 31 years old (ranged from 19 to 53 years). Based on the Tile classification,17 cases were type CI and 12 case were type C2. Tornetta standard and Majeed score were used to evaluated the clinical effect after operation.

RESULTSAll patients were followed up from 10 to 24 months with an average of 16 months. There were no nerve injuries and other organ injuries, only one case of infection was found and it was cured with wound dressing. Time of fracture union was from 14 to 18 weeks with the mean of 16.2 weeks. No loosening, slippage or breakage of the screw were found. According to the Tornetta standards,14 cases obtained excellent results, 10 good, 4 fair and 1 poor. The Majeed score was 87.2 ± 11.3, 16 cases got excellent results, 9 good and 4 fair.

CONCLUSIONClosed reduction and screw internal fixation combined with external fixation is an effective way to treat unstable pelvic fractures with Tile type C1 and Tile type C2.

Adolescent ; Adult ; Child ; External Fixators ; Female ; Fracture Fixation, Internal ; methods ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Pelvic Bones ; injuries ; surgery

Objective To establish a method of reverse hybridization to detect five subfamilies of low risk Human Papillomaviruses(HPV6,11,42,43 and 44)and eighteen subfamilies of high risk HPV (HPV16,18,31,33,35,39,45,51,52,53,56,58,59,66,68,73,83 and MM4)in one reaction.Methods Special probes for twenty-three HPV subfamilies were fixed on nylon membrane bars,biotin labeled general primers mediated polymerase chain reaction(GP-PCR)were applied in HPV DNA amplification.PCR amplified DNA fragments were reversely hybridized with special probes that were fixed on the membranes. All samples(136)detected by reverse hybridization method were paralleled with the methods of Hybridization Capture Ⅱ(HC-Ⅱ)and sequencing.Results Positive rate of the 136 samples detected by reverse hybridization was 41.9%,while HC-Ⅱ 42.6% and sequencing 40.4%.Reverse hybridization detection indicated coherence with the other two methods(Kappa 0.8644 and 0.9089,respectively).While sequencing was lab standard for DNA test,the sensitivity was 96.36%,specificity was 95.06%,accuracy was 95.59%.Conclusions Method of reverse hybridization is adaptable to 23 kinds of HPV subfamilies, which can confirm the exactly subfamilies of HPV infection.This method is adaptable in clinical detection of HPV,with high sensitivity,high specificity,simply and convenient operation and the results are easily to be read.

A high Streptolydigin-producing strain Streptomyces lydicus AS 4.2501-L8 with genetic stability was obtained from the original strain S. lydicus AS 4.2501-P28 by mutagenesis with UV and acridine orange, followed by resistance selection. Its yield of Streptolydigin was 177.0 ?g/mL, 96.2% higher than the original strain.

Objective To examine the changes in the expression of voltage-gated sodium channel?subunit mRNA in the dorsal root ganglion(DRG)and the role it plays in the neuropathie pain.Methods Thirty- two male SD rats weighing 250-400g were randomly divided into 2 groups:groupⅠneuropathic pain(SNL,n= 20)and groupⅡsham operation(n=12).Neuropathic pain was produced by ligation of right sciatic nerve according to Seltzer.Paw withdrawal latency to noxious thermal(PWHL)and mechanical(PWML)stimulation were measured before(baseline)and 1,2,3,5,8,11,14,28 day after sciatic nerve ligation(SNL).DRG at L_(4,5) was isolated on the 14th day after SNL in 8 SNL and 4 sham-operated animals for determination of sodium channel?subunit mRNA expression(by in-situ hybridization).Results PWHL and PWML were significantly decreased on the 2nd-28th day after SNL as compared to the baseline in SNL group.There was no significant difference in?_1 subunit mRNA expression between the 2 groups.The?_2 subunit mRNA expression in DRG was hardly detectable.The?_3 subunit mRNA expression in DRG on the operated side was significantly higher in SNL group than in sham-operation group.Conclusion The up-regulation of sodium channel?_3 subunit mRNA expression in DRG may play an important role in neuropathic pain.

Objective To clarify the cellular localization of triptolide and to explore its in-cell action sites.Methods 4-(Bro-momethyl)-7-methoxycoumarin was employed to label triptolide,then labelled triptolide was incubated with human hepatoma carci-noma cells.Subsequently,incubated cells were subjected to stain with fluorescent dye DiI or PI,which were specific to cytoplasmic membrane system and nucleus,respectively.Results Compared with the non-triptolide control,coumarin labelled triptolide shown a light blue fluorescence under UV excitation;Co-localization with DiI showed that triptolide exist in cytoplasm and(or)on cell mem-brane;Co-localization with PI showed that triptolide located in cell nucleus.Moreover,microscopic observation indicated that the fluorescence intensity in nucleus was denser than that in cytoplasm.Conclusion The presnt study demonstrate that triptolide main-ly act in nucleus,followed by acting in cytoplasm and(or)on cell membrane.