Objective To construct a recombinant interfering RNA(miRNA)plasmid vector targeting annexinA3 and observe its impact on the growth and apoptosis of human gallbladder cancer cell line SGC-996.Methods Four small fragments of miRNA and one negative control were sequenced and cloned into vector pcDNATM 6.2-GW/EmGFPmiR.The recombinant plasmids were then transfected into gallbladder cancer cell line SGC-996 by positive liposomal transfection.The transcription and translation level of annexinA3 expression were detected by RT-PCR and Western Blot.Meanwhile, the proliferation and apoptosis of transfected tumor cells were determined by MTT and flow cytometry.Results The recombinant plasmids containing annexinA3 miRNA were successfully constructed.AnnexinA3 expression was significantly knocked down after miRNA plasmid transfection.Transfection of annexinA3 gene miRNA could effectively inhibit the proliferation of gallbladder cancer cell and promote apoptosis.Conclusion AnnexinA3 gene silenced by miRNA can induce the apoptosis of human gallbladder cancer cell line SGC-996.The process might serve as a potential approach for cancer gene therapy.

BACKGROUND:Cervical decompression and fusion internal fixation wil accelerate adjacent segment disc degeneration, and it is not clear whether single segment instaibility can increase the adjacent segment disc degeneration. OBJECTIVE:To study the changes of morphology, proteoglycan and col agen type Ⅱ in the adjacent intervertebral disc of the cervical instability models. METHODS:Sixteen New Zealand white rabbits were divided into two groups randomly, with eight rabbits in the control group and eight rabbits in the model group. The animal cervical instability models were made by destroyed partly annulus fibrosus and absorbed C 5/6 nucleus pulposus through anterior cervical puncture. After 12 weeks, the animal models were tested by X-ray film. Al rabbits were sacrificed and 10 mg nucleus pulposus of the intervertebral discs of C 4/5 cut from sagittal plane were harvested and stored under 0 ℃. The content of proteoglycan in nucleus pulposus was tested with phloroglucinol method. Then, the paraffin sections of intervertebral disc tissues were taken for hematoxylin-eosin staining and SABC immunohistochemical staining. RESULTS AND CONCLUSION:The notochord cells of C4/5 intervertebral discs in the experimental group was decreased, and being replaced by fibroblast-like cells. Round chondrocytes could be seen occasional y and intervertebral discs annulus fibrosus became rough and arranged disorderly, the hyaline degeneration and pigmentation were observed as wel as the fibrochondrocytes, and there was a gap between inner and outer annulus fibrosus. The content of proteoglycan was decreased in the nucleus pulposus, and there was significant difference between two groups. The col agen type Ⅱ in the degenerative disc nucleus pulposus and annulus fibrosus of the experimental group was lower than that of the control group. Cervical instability can lead to adjacent intervertebral disc degeneration with the morphological changes and decreased content of proteoglycan and col agen type Ⅱ.

Objective To observe the effects of butylphthalide on the expression of autophagy-related protein and mRNA in l-meth-yl-4-phenyl-pyridiniumion (MPP+)-induced SH-SY5Y cells, and to explore the protective effect and possible mechanism of butylphthalide to the cell model of Parkinson's disease. Methods The SH-SY5Y cells were divided into control group (A), MPP+group (B), rapamycin pre-treated+MPP+group (C) and Butylphthalide pretreated+MPP+group (D). The relative viability of SH-SY5Y cells induced by MPP+was measured with MTT assay, the morphology of SH-SY5Y cells was observed. The expression of microtubule associated protein 1 light chain 3 (LC3)-II/I and Beclin 1 protein was detected by Western blotting. And the expression of LC3-II/I and Beclin 1 mRNA were assayed by re-al-time quantitative reverse transcription-PCR (RT-PCR). Results The viability rates of cells were significantly lower in group B than in group A (t=20.270, P<0.001), and were significantly higher in groups C and D than in group B (t>8.770, P<0.001), however, there was no significantly difference between groups C and D (t=2.270, P=0.064). The expression of LC3-II/I and Beclin 1 was higher in group B than in group A (t>6.647, P<0.01), and was higher in groups C and D than in group B (t>3.630, P<0.01), however, there was no significantly differ-ence between groups C and D (t<2.238, P≥0.05). Conclusion Butylphthalide could prevent the injury of SH-SY5Y cells induced by MPP+, which may affect Parkinson's disease by inducing autophagy.

BACKGROUND:Pelvic and acetabular fractures in elderly are frequently observed in the clinic, and the incidence gradual y increased. Moreover, it is difficult to conduct reduction and fixation due to their physical status and osteoporosis to different degrees. Locking plate is ideal fixation material in surgical treatment of acetabular fractures in elderly due to the special functions and good molding of screw and locking plate.
OBJECTIVE:To analyze the advantages and disadvantages of locking reconstruction plate used in the acetabular fracture for the aged patients and to improve the clinical effect of acetabular fractures in elderly.
METHODS:From January 2010 to February 2013, 21 aged patients with acetabular fracture were treated by open reduction and internal fixation with locking reconstruction plates made of titanium and 00Cr 18 Ni 14 Mo 3 stainless steel in the Department of Orthopedics Affiliated Hospital of Jining Medical University.
RESULTS AND CONCLUSION:Of 21 patients, 20 cases were fol owed up for 12-18 months (mean 14 months), but one dropped out. Al the fractures were healed within 4-6 months and no failure cases. According to Matta evaluation criteria, the satisfactory rate was 90%. According to the D’Aubigne scoring system, the excellent and good rate was 95%. These data indicated that locking reconstruction titanium plate is reliable, has low failure rate and satisfactory clinical effects for acetabular fracture in aged patients, and is ideal fixation material in treatment of acetabular fractures in elderly.

A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the determination of butoconazole in human plasma. Human plasma samples of 0.2 μL were pretreated by a single step protein precipitation procedure and analyzed using a high performance liquid chromatography (HPLC) electrospray tandem mass spectrometer system. The compounds were eluted isocratically on an Inertsil ODS-SP column (100 mm×2.1 mm, 3 μm), ionized using a positive ion atmospheric pressure electrospray ionization source and analyzed using multiple reaction monitoring (MRM) mode. The ion transitions monitored were m/z 412.8→165.1 for butoconazole and m/z 453.4→230.3 for the internal standard. The chromatographic run time was 3.5 min per injection, with retention time of 2.47 min and 2.15 min for butoconazole and repaglinide, respectively. The method was validated to be linear over the range of 20 to 8000 pg/mL (r>0.999) by using a weighted (1/x(2)) quadratic regression. The mean recovery rate was more than 86.7%, and the intra- and inter-day precision of the quality control samples (QCs) was less than 8.3% and the accuracy ranged from 96.0% to 110.2%, which indicated that the quantitative method was reliable and accurate. The method is simple, rapid, and has been applied successfully to a pharmacokinetics study of butoconazole nitrate suppositories in healthy Chinese females.

Abstract: Objective - - To establish a pre column derivatization high performance liquid chromatography method for detecting Methods dimethyl sulfate (DMS) in workplace air. DMS in workplace air was collected with mercaptopyridine impregnated （ silicone tube. The derivative of DMS and mercaptopyridine was eluted by mobile phase phase A: water, phase B: acetonitrile, ∶ the volume ratio was 40 60) , and separated with a C18 column, then detected with diode array detector and quantitated by a Results - standard curve. The linear range of DMS was 0.17 40.00 mg/L, with the correlation coefficient of 0.999 95. The detection limit and the lower limit of quantitation were 0.05 and 0.17 mg/L respectively. The minimum detection concentration and minimum quantitation concentration were 0.02 and 0.04 mg/m³, respectively (air sample volume of 4.5 L, 1.0 mL sample - - - solution). The average desorption efficiency was 98.40% 102.00%. The within run and between run relative standard deviations - - were 0.61% 3.92% and 1.71% 6.00%, respectively. The samples could be stored at room temperature for at least 14 days. Conclusion This method can be used to detect DMS in workplace air.

Abstract: Nitrobenzene compounds (NBCs) are widely used in the world. It has 40 isomers such as nitrobenzene, dinitrobenzene and nitrotoluene, that are highly toxic and difficult to degrade and can cause harm to human health in different degrees. At pres⁃ ent, there is no unified standard method and occupational exposure limit for the detection of NBCs in the air. In terms of sampling medium, solid adsorption tube is mostly used for trapping vapor state NBCs, and filter membrane and solid adsorption tube are mostly used in series for sampling coexist NBCs in vapor state and aerosol state. In the detection methods, gas chromatography and liquid chromatography are common, and ultraviolet spectrophotometry, Raman spectroscopy, ion migration spectrometry and some other rapid response methods and technologies are also used in the detection of NBCs. In the detection of NBCs by gas chro⁃ matography, capillary column separation is commonly used, and the main detectors are flame ionization detector, electron capture detector and mass spectrometry detector. It is of practical significance to establish a method with high sensitivity, strong practica⁃ bility, convenient operation, and can simultaneously collect and detect a variety of NBCs in different states.

The prescriptions including Polygoni Multiflori Caulis that built by Pro. Yan were collected to build a database based on traditional Chinese medicine (TCM) inheritance assist system. The method of association rules with apriori algorithm was used to achieve frequency of single medicine, frequency of drug combinations, association rules between drugs and core drug combinations. The datamining results indicated that in the prescriptions that including Polygoni Multiflori Caulis, the highest frequency used drugs were parched Ziziphi Spinosae Semen, Ostreae Concha, Ossis Mastodi Fossilia, Salviae Miltiorrhizae Radix Et Rhizoma, Paeoniae Rubra Radix, and so on. The most frequent drug combinations were "Polygoni Multiflori Caulis-parched Ziziphi Spinosae Semen", "Ostreae Concha-Polygoni Multiflori Caulis", and "Polygoni Multiflori Caulis-Ossis Mastodi Fossilia". The drug association rules of confidence coefficient 1 were "Ostreae Concha-->Polygoni Multiflori Caulis", "Poria-->Polygoni Multiflori Caulis", "parched Ziziphi Spinosae Semen-->Polygoni Multiflori Caulis", and "Paeoniae Alba Radix-->Polygoni Multiflori Caulis". The core drug combinations in the treatment of insomnia were Ossis Mastodi Fossilia, Polygoni Multiflori Caulis, Salviae Miltiorrhizae Radix et Rhizoma, Ostreae Concha, Polygalae Radix, Margaritifera Concha, Poria, and parched Ziziphi Spinosae Semen. And the core drug combinations in the treatment of obstruction of Qi in chest were Salviae Miltiorrhizae Radix Et Rhizoma, Polygoni Multiflori Caulis, parched Ziziphi Spinosae Semen, Trichosanthis Fructus, Allii Macrostemonis Bulbus, and Paeoniae Rubra Radix.
Data Mining ; methods ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Humans ; Medicine, Chinese Traditional ; methods ; Plant Stems ; chemistry ; Polygonaceae ; chemistry ; Prescriptions

Analyzed the prescriptions for phlegm retention syndrome that built by Ma Peizhi by the association rules and clustering algorithm, the frequency of drug usage and the relationship between drugs could be get. And from that we could conclude the experiences for phlegm retention syndrome of Ma Peizhi of menghe medical genre. The results of the analysis were that 18 core combinations were dig out, such as Citri Exocarpium Rubrum-Eriobotryae Folium-Citri Reticulatae Pericarpium. And there were 9 new prescriptions were found out such as Aurantii Fructus-Citri Exocarpium Rubium-Eriobotryae Folium-Citri Reticulatae Pericarpium. The results of the analysis were proved that Ma Peizhi of Menghe Medical Genre was good at curing phlegm retention syndrome by using the traditional Chinese medicine of mild and light, such as the medicines of mild tonification, and clearing damp and promoting diuresis.
Algorithms ; Data Mining ; Databases, Factual ; Drug Prescriptions ; standards ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Mucus ; chemistry ; Practice Patterns, Physicians' ; Respiratory Tract Diseases ; drug therapy

The clinical prescriptions of hemoptysis that built by Menghe physician Ma Peizhi were collected, and were analyzed by the method of unsupervised data mining, such as association rules and clustering algorithm. From the prescriptions, we got the frequency of drugs, the association rules among drugs, 10 core drug combinations and 5 new prescriptions. Accordingly, after the analysis, Menghe physician Ma Peizhi had rich experience in the treatment of hemoptysis. His therapy for hemoptysis was clearing heat and moistening lung, dispel heat from blood to stop bleeding, and enriching yin and blood. And we can also make a conclusion that the TCM inheritance support system is of great practical value for minning the old doctors' clinical experiences.
Data Mining ; Databases, Factual ; Drug Prescriptions ; standards ; Drugs, Chinese Herbal ; chemistry ; therapeutic use ; Hemoptysis ; drug therapy ; Humans ; Practice Patterns, Physicians'