Objective To investigate the influence of different diluents(physiologic saline, dis-tilled water and human inactivated serum) on measurement of 15 items of biochemical parameters.Methods Fifteen items of biochemical parameters [alanine transaminase(ALT), aspartic transaminase (AST), alkaline phosphatase( ALP), gamma glutamyl transpeptidase ( GGT), creatine kinase ( CK),lactate dehydrogenase(LD), hydroxybutyrate dehydrogenase(HBDH), total bilirubin(TBIL), direct bilirubin(DBIL), total bile acid(TBA), ereatinine(Cr), uric acid(UA), eholesterol(CHO), glucose (GLU), and blood urea nitrogen(BUN)] were chosen. For each parameter, 45 serum samples with different eoncentrations of the parameter were collected. After diluted with different diluents(physio-logic saline, distilled water or human inactivated serum), the serum samples were detected by applying the fully automated biochemical analyzer. The mean value was calculated and statistical analysis was performed. Results There were some differences of detection results when the specimens were diluted with different diluents. ALT, AST, GGT, DBIL, and HBDH serum samples could be diluted by 10 times with physiologic saline, distilled water or human inactivated serums ALP and TBA serum sam-ples could only be diluted with inactivated serum, otherwise its result would be lower; GLU, TBIL samples could be diluted with distilled water and inactivated serums for BUN, CR, UA, CK, LDH,and CHO samples, physiologic saline or human inactivated serum might be optimal; if distilled water was chosen, the results of other parameters tented to decline except UA. It was BUN was improper to dilute the BUN samples with distilled water. In addition, there was no significant difference between the items diluted by 5 times and 10 times with physiologic saline. All the 15 items could be diluted with inactivated serum. Conclusion The inactivated serum should be the first choice of diluents to e-nure the accurate results of biochemical parameters. If the prepared inactivated serum is absent, we may choose other diluents according to the above-mentioned results.

Objective To study the prevention of gentamicin ototoxicity by resveratrol(Res) in guinea pigs.Methods Auditory brainstem response and cochlear preparation transmission electron microscope were used to evaluate the effects of hair cell on hearing threshold and morphology of hair cell. Serum levels of GM were also tested. It was comprised of five groups,each group had eight animals.GM and Res were tested alone and in combination,besides control group.Results GM group developed a progressive threshold shift. Injury in high frequency was significantly more severe than that in low frequency (P

Objective To establish a reversed-phase high performance liquid chromatograph ( RP-HPLC ) method for determination of equilibrium solubility and oil/water partition coeficient of phloridzin in different solvents. Methods A RP-HPLC method was established to detect the concentration of phloridzin in water and different organic solvents. The partition coefficients in the n-octanol-water/buffer solution systems of phloridzin were determined by shaking flask method. The Inertsil ODS-3 (4.6 mm×150 mm, 5μm) column was used and the detection wavelength was 284 nm.The flow rate was 1.0 mL??min-1, and acetonitrile-0.05% phosphoric acid(30??70)was used as mobile phase. Results The equilibrium solubility of phloridzin was 2.07 mg??mL-1 in water and 838.63 mg??mL-1 in methanol at 25 ℃.A good linear relationship of phloridzin was obtained within the range of 0.054 9-1.098 0 μg.The regression equation was Y=2 152.9X+7.26 (r=0.999 9).The solubility values of phloridzin were higher in ethanol and propylene glycol than in other solvents. Conclusion RP-HPLC method is simple, quick and accurate for the determination of phloridzin.Phloridzin was almost insoluble in petroleum ether and poorly soluble in water.The equilibrium solubility is higher in methanol than in other solvents. The apparent distribution coefficient of phloridzin varies significantly with pH under the alkaline conditions but less in the acidic solution.

BACKGROUND:At present, a variety of extracel ular matrix-derived scaffolds have been successful y applied for cartilage tissue engineering in experiment and clinical practice. OBJECTIVE:To summarize the application and research status of extracel ular matrix-derived scaffolds in cartilage tissue engineering. METHODS:A computer-based online search in PubMed, CNKI, CqVip and WanFang databases was performed using the keywords of“tissue engineering, cartilage, extracel ular matrix, scaffolds”in English and Chinese, respectively. A total of 1 140 literatures were retrieved, and final y 65 eligible literatures were included. RESULTS AND CONCLUSION:In terms of the components, extracel ular matrix-derived scaffolds are divided into monomeric natural polymers, mixed natural polymers, natural polymers compositing with synthetic polymers as wel as acel ular extracel ular matrix-derived materials. Extracel ular matrix-derived scaffolds hold good biocompatibility and degradability, and can promote proliferation and differentiation of choncrodytes;therefore, they as good bionic scaffolds have been applied for cartilage tissue engineering in clinical practice, However, poor mechanical properties and difficulty to molding should never be ignored. Further research should focus on improving the preparation technology by combining synthetic materials with extracel ular matrix-derived scaffolds for cartilage tissue engineering.

BACKGROUND:Accumulative evidence supports that co-culture technology can be applied to construct the tissue-engineered cartilage with excellent biological characters. OBJECTIVE:To elaborate the co-culture concept and conclude and analyze seed cell sources, cel mixed ratio, spatial y-defined co-culture models and biomaterials in co-culture systems to conclude and analyze the biological characters of tissue-engineered cartilage, and to prospect progression of co-culture systems in cartilage tissue engineering. METHODS:The first author retrieved the databases of PubMed, Web of Science, and CNKI for relative papers published from January 1976 to May 2016 using the keywords ofco-culture, co-culture systems;articular cartilage, chondrocytes, mesenchymal stem cells;tissue engineering, articular cartilage tissue engineeringin English and Chinese, respectively. Finally 60 literatures were included in result analysis, including 1 Chinese and 59 English articles. RESULTS AND CONCLUSION:Co-culture technology emphasizes the role of microenvironment in terms of various physical, chemical and biological factors in the cell processing. In cartilage tissue engineering, co-culture systems contribute to maintain the viability and natural cell phenotype of chondrocytes and induce cartilage differentiation of mesenchymal stem cells. In addition, co-culture technology provides a novel way for cartilage tissue engineering to overcome the shortage of chondrocytes and repair injury to the cartilage-subchondral bone. However, the mechanisms of cell-cell interaction in co-culture systems still need to be explored in depth, so as to optimize the co-culturing conditions and construct perfect tissue-engineered cartilage.

Objective To deepen the understanding of chronic disseminated candidiasis(CDC)in patients with acute leukemia(AL).Methods CDC was investigated in 119 AL patients who received induction chemotherapy from August 2004 to May 2005.Clinical manifestations,laboratory tests,imaging modalities,diagnosis and treatment were investigated retrospectively.Results Three patients(2.5%) were identified to be suffering from CDC.All the three patients had an absolute neutrophil count (ANC)＜0.5 × 109/L for more than 15 days.Two patients had normal ANC when they were diagnosed to have CDC.The common manifestations in these three patients were persistent fever,splenohepatomegalia and percussion pain in hepatic region.Meanwhile,2 of them were accompanied with cough,expectoration and dyspnoea.The abnormal laboratory test observed during the course of infection in two of them was increase of alkaline phosphatase.Computed tomography scan showed multiple hypodense lesions in the liver and spleen in all the three patients:two of them showed multiple nodular patchy shadOW$in lungs.Nuclear magnetic resonance imaging showed multiple abnormal signal in liver,spleen and kidneys in one of the patients.Two patients had positive bleed fungal cultures and histologic examination in one of the patients were positive for Candida tropicalis.Two patients received amphotericin B therapy empirically,but it was replaced by amphotericin B colloid dispersion (ABCD) later in one and combined with voficonazole in another because of unresponsiveness to the drug.One patient took a favorable turn after receiving ABCD therapy for 45 d,which was replaced by voriconazole because of the emergence of fever after disconfinuation of ABCD.All the three patients received further chemotherapy smoothly after the diagnosis of CDC.Conclusion The diagnosis of CDC remains difficult.Fungal blood cultares and histologic examination have been considered in many studies as the golden standard for the diagnosis of CDC.Amphotericin B is the cornerstone of treatment in patients with CDC and lipid formulations of amphotericin B can be used in CDC patients who are intolerant of or refractory to conventional amphotericin B.Voriconazole has a favorable response for refrectory/relapse patients and could be used for second line trectment.The development of CDC in patients with acute leukemia does not preclude further chemotherapy.

Objective To set up a multiplex real time quantitative PCR method to detect the expression of WT1 and MDR1 gene simultaneously in acute leukemia patient. Methods Total RNA was extracted from k562 cell line and was reverse transcribed to cDNA by the outer primers of WT1 and MDR1 respectively. The cDNA of WT1 and MDR1 were purified and digested by Bam H Ⅰ and Bgl Ⅱ , and then the two fragments were ligated to form the recombinant fragment WT1 + MDR1. The outer forward primer of WT1 and outer reverse primer of MDR1 were used to amplify the recombinant fragment WT1 + MDR1. The PCR product was purified and cloned into pMD18-T vector, and then transferred into E. coli DH-5α. A new kind of WT1-MDRl-contained standard plasmid was obtained from the positive colony. The recombinant plasmid was verified by digestion with restriction enzyme and PCR amplification. A multiplex real time quantitative PCR method was set up with FAM-labeled MDR1 probe and VIC-labeled WT1 probe in one reaction tube. The WT1 and MDR1 gene expression was detected in forty-seven AL patients and thirty-two controls by this method. Seven patients were followed-up to elucidate the relationship between the gene expression levels and clinical prognosis. Results The recombinant plasmid was confirmed by EcoR1 digestion and PCR amplification. The multiplex real time quantitative PCR technique could reach the sensitivity of WT1 and MDR1 gene up to 102 copy/μl. The standard curve slopes were 0. 999 and 0. 998. The WT1 [ 37 000( 163-6 370 000 )copies/μg RNA ] and MDR1 [ 76 200( 179-18 000 000 )copies/μg RNA ]expression levels of AL patients were significantly higher as compared to the controls [ 258( 0-643 ) copies/μg RNA and 333( 0-779 )copies/μg RNA ]( Z= 6. 755,6. 736, P ＜ 0. 01 ). Following up seven patients with similar regimen of chemotherapy, the WT1 and MDR1 expression correlated to the clinical course. Three AL patients with WT1 and MDR1 expression levels (2 170 and 86 900, 1 130 and 5 860, 1 170 and 586 copies/μg RNA )significantly decreased after chemotherapy and kept in the low range ( 370 and 560,138 and 980, 150 and 690 copies/μg RNA ), and had a favorable outcome. Three AL patients with WT1 and MDR1 expression levels ( 1 600 and 11 800, 24 800 and 968, 48 200 and 1 100 000 copies/μg RNA )decreased after initial chemotherapy, but increased significantly afterwards (20 314 and 25 660,184 364 and 31 530, 15 680 and 878 000 copies/μg RNA ),and suffered clinical relapse. One patient with high WT1 and MDR1 expression levels ( from 81 600 and 1 200 000 copies/μg RNA to 124 100 and 7 632 400 copies/μg RNA )showed the persistence of disease. Conclusions A multiplex real time quantitative PCR method to detect WT1 and MDR1 gene simultaneously is constructed successfully. The expression of WT1 and MDR1 may provide useful information for AL patients prognosis.

This study was aimed to identify and distinguish Metacordyceps liangshanensis recorded by the Sichuan Province Traditional Chinese Medicine Standard from its adulterants and its relative species by combining ITS and COI barcode sequences in order to study the feasibility of this new method. After extracting DNA of 28 species of Cordyceps samples, DNA were amplified and sequenced. And then, ITS and COI sequences were received. Codon-Code Aligner V3.7.1 and Mega 5.0 were used to analyze the variable site and construct the N-J tree. The results showed that the minimum ITS inter-specific K-2P distance was relatively higher than the maximum intra-specific K-2P distance. The inter-specific sequence divergence between M. liangshanensis and its adulterants exhibited high while intra-specific sequence divergence exhibited low. And COI one was the same case. N-J tree of both ITS and COI indicated that same genus belonged together and each species belonged to relatively independent branch. It was concluded that based on the ITS and COI gene, the technology of DNA barcode can be an excellent identification of M. liangshanensis, its adulterants and its relative species. It provided technical support for the further research on species molecular identification and phylogenetics of Cordyceps .

Objective To explore the effects of application of self-made pipe protective belt for catheter of thrombolysis on treatment of patients with lower limb ischaemia via femoral artery puncture. Methods From March 2016 to December 2016,116 patients with lower limb ischaemia treated with femoral artery puncture in a tertiary hospital in Hebei Province were recruited using convenience sampling method. We divided all patients into the ex-perimental group (58 cases) and the control group (58 cases) according to random number table method. For the experimental group,we used sterile transparent dressings to fix the catheter of thrombolysis,then applied self-made pipe protective belt to fix it.In the control group,we used traditional sterile transparent dressings to fix the catheter of thrombolysis,then used self-adhesive elastic bandage for external fixation by cross overlapping. Incidence rate of accidental extubation,pressure ulcer related to medical equipment (tee joint) and medical adhesive-related skin in-jury from two groups were observed. Results The incidence rate of accidental extubation in the experimental group was lower than that in the control group (P<0.01);there were statistically significant differences in the pressure ul-cer related to medical equipment (tee joint)and medical adhesive-related skin injury between two groups (P<0.05). Conclusion During catheter-directed thrombolysis for patients with lower limb ischaemia via femoral artery punc-ture,the self-made pipe protective belt can reduce the incidence of accidental extubation,avoid adverse events such as related to medical equipment (tee joint)and medical adhesive-related skin injury effectively,and ensure safety of the patients.

BACKGROUND: The researches of Pulling and Rotating Oblique Pulling Manipulation focus on clinical research and biomechanical changes of lumbar functional units, and the mechanical characteristics of manipulation are little reported.OBJECTIVE: To study the characteristics of the Pulling and Rotating Oblique Pulling Manipulation by digital method, so as to provide quantitative basis for inheritance, teaching and learning, promotion, and basic research of the manipulation.METHODS: The force-time curve and the force of the manipulators were tested and recorded with the multi-point membrane pressure measurement system. The kinetic parameters (the average load force, the average minimum force and the maximum impact force) were shown. The values of = the flip time, flip speed and impulse were measured by the test analysis system.RESULTS AND CONCLUSION: (1) The average load force was (145.86±34.80) N, duration was (1.43±0.46) s, the average minimum force was (72.24±13.87) N, the maximum impact force was (446.21±143.98) N, the flip time was (0.55±0.15) s, flip speed was (914.52±259.18) N/s, and the impulse was (256.21±82.30) N?s. (2) The rising slope of the impulse was (93.96±6.94), and the falling slope was (-82.70±26.10). (3) To conclude, the characteristics of manipulation from hand are analyzed in views of digitization, which provides an objective evaluation index for Lin's manipulation.