AIM: To establish the method for quality control of Zangfukang Supposit ory (Radix Przewalskii tanguticae, Pericarpium Zanthoxyli, Herba Oxytropis falca tae, etc.). METHODS: Radix Przewalskii tanguticae, Pericarpium Zanthoxyli, Her ba Oxytropis falcatae were identified by TLC, and the content of hyoscyamine in Radix Przewalskii tanguticae was determined by HPLC. RESULTS: The ethyl acetate-methanol-ammonia(17∶2∶1) was applied as the developer for the identification of Radix Przewalskii tanguticae on silica gel G coating plate. The cyclohexane-ethyl acetate-formic acid (30∶15∶1) was applied as the developer for the identification of Pericarpium Zanthoxyli on s ilic a gel G coating plate. The cyclohexane-ethyl acetate-formic acid (50∶10∶1) w as taken as the developer for the identification of Herba Oxytropis falcatae als o on silica gel G coating plate. Hyoscyamine showed a good linear relationship a t a range of 2.5～25?g,r=0.9999. The average recovery was 99.59%, and RSD was 1.34%. CONCLUSION: The methods are accurate and reliable, and can control the quality of this preparation effectively.

cases of vitreous haemorrhage secondary to Eales′ disease were selected for vitrectomy. According to the duration of vitreous haemorrhage, the patients were divided into two groups : Group 1 (20 eyes)-early vitrectomy group with a duration between 3 to 6 months; Group 2(20 eyes)-deferred vitrectomy group with a duration of more than 6 months . All the patients were followed up for a minimum period of 3 months following vitrectomy. The eyes in Group 1 showed a preoperative ultrasonic picture of complete posterior detachment, a final visual acuity of 0.6 or better in 13(65%) was achieved in mild vitreous organization on kinetic echography. Poor visual outcome in the deferred group was secondary to cystoid macular oedema, macular scar, macular pucker formation and macular degeneration. Improved visual outcome in the early vitrectomy group is probably due to haemorrhage and its products which have no time to damage the macula and to advance into macular traction and cystoid macular oedema.

Objective To observe the results of visual functional MRI(fMRI)of injury side with those of uninjured side,in patients with visual field defects and retrochiasmastic tumor.The potential clinical usefulness of visual fMRI data during functional examination on visual centre,defining the tumor field and surgery,was evaluated in patients with occipital lobe tumour.Design Case series.Participants Two patients with occipital lobe tumor and homonymous visual field defects,injury sides contrast with uninjured ones.Methods Patients were studied with fMRI using monocular checker stimulation(5Hz).The data obtained were analyzed with statistical parametric maps software(P

AIM: To establish the method for quality control of Gutongning Liniment(Herba Epimedii,Semen Strychni,Herba Ephedrae,etc.) METHODS: Herba Ephedrae,Radix Notoginseng,Ramulus Cinnamomi,Flos Caryophylli,Flos Daturae were identified by TLC,icariin content in Herba Epimedii and strychnine content in Semen Strychni were determined by HPLC. RESULTS: The identified characteristics of chromatography were distinct and the spots were clear.Icariin showed a good linear relationship at a range of 0.2-1.2 ?g,r=0.999 8.The(average) recovery was 10.3%,and RSD was 1.5%.Strychnine showed a good linear relationship at a range of 0.8-4.8 ?g,r=0.999 7.The average recovery was 99.17%,and RSD was 1.4%. CONCLUSION: The methods are accurate and reliable,and can be used to control the quality of this preparation effectively.

Objective To determine the effect of different serum insulin-like growth factor-Ⅰ(IGF-1)levels on mouse cancer.Methods A total of 120 male mice at 6 weeks of age(60 control mice and 60 LID mice)were subcutaneously injected colon tumor CT26 cell line.Each group was random divided into two subgroups respectively,every 10 mice of one subgroup were injected subcutaneously with growth hormone(GH)(1ms/kg)daily from the 10th,14th and18th days respectively until the 22nd days,and the other subgroup received saline injection.Results All mice treated with GH have higher level of IGF-1,compared with those treated with saline.High level of IGF-1 promoted the development of cachexia in these mice treated with GH from the 10th days.However,the level of IGF-1 has negative correlation with the cancer cachexia state for mice treated with GH from the 14th days.Conclusion Circulating IGF-1 and GH play an important role in tumor growth 4nd cachexia development in the early stage of cancer and can ameliorate the state of cachexia in the advanced stage.

Objective To clone and express of Rv0091 encoding protein in Mycobacterium tuberculosis,identify and characterize of the enzyme activities.Methods Construct the Rv0091 prokaryotic expression plasmid,the vector was transformed into E.coli strain BL21trxB.After induced by IPTG,recombinant protein was purified by Ni2+-NTA chromatography and analyzed for purity by SDS-PAGE gels stained with Coomassie Blue.Immunological activity was identified by Western blot.The recombinant protein molecular weight was identified by Mass spectrometry.The enzyme-coupled assay detectes enzyme activity.Results The expression plasmid pET32a-Rv0091 was constructed and expressed in E.coli.BL21trxB,and the optimum expression system was conformed.The purity of the recombinant protein was more than 95％.Western blot analysis confirmed that recombinant protein was one of Mycobacterium tuberculosis proteins.Mass spectrometry identified the relative molecular weight and theoretical molecular weight was basically the same.Enzyme assay showed the recombinant protein able to catalyze the substrate MTA.Enzymatic properties showed that the optimal buffer for the phosphate and Hepes buffer,the poor thermal stability of the enzyme,the optimal temperature of 37℃,optimal pH10-12,when the pH ≤7,the protein denaturation and loss of some vitality.Conclusion The recombinant protein methylthioadenosine nucleosidase(MTAN) was obtained and enzyme activity was detected and plays a key role in the metabolism of Mycobacterium tuberculosis.

OBJECTIVETo appraise the significance of extravascular lung water index (EVLWI), pulmonary vascular permeability index (PVPI), and intrathoracic blood volume index (ITBVI) in the differential diagnosis of the type of burn-induced pulmonary edema.

METHODSThe clinical data of 38 patients, with severe burn hospitalized in our burn ICU from December 2011 to September 2014 suffering from the complication of pulmonary edema within one week post burn and treated with mechanical ventilation accompanied by pulse contour cardiac output monitoring, were retrospectively analyzed. The patients were divided into lung injury group ( L, n = 17) and hydrostatic group (H, n = 21) according to the diagnosis of pulmonary edema. EVLWI, PVPI, ITBVI, oxygenation index, and lung injury score ( LIS) were compared between two groups, and the correlations among the former four indexes and the correlations between each of the former three indexes and types of pulmonary edema were analyzed. Data were processed with t test, chi-square test, Mann-Whitney U test, Pearson correlation test, and accuracy test [receiver operating characteristic (ROC) curve].

RESULTSThere was no statistically significant difference in EVLWI between group L and group H, respectively (12.9 ± 3.1) and (12.1 ± 2.1) mL/kg, U = 159.5, P > 0.05. The PVPI and LIS of patients in group L were respectively 2.6 ± 0.5 and (2.1 ± 0.6) points, and they were significantly higher than those in group H [1.4 ± 0.3 and (1.0 ± 0.6) points, with U values respectively 4.5 and 36.5, P values below 0.01]. The ITBVI and oxygenation index of patients in group L were respectively (911 197) mL/m2 and (136 ± 69) mmHg (1 mmHg = 0.133 kPa), which were significantly lower than those in group H [(1,305 ± 168) mL/m2 and (212 ± 60) mmHg, with U values respectively 21.5 and 70.5, P values below 0.01]. In group L, there was obviously positive correlation between EVLWI and PVPI, or EVLWI and ITBVI (with r values respectively 0.553 and 0.807, P < 0.05 or P < 0.01), and there was obviously negative correlation between oxygenation index and EVLWI, or oxygenation index and PVPI (with r values respectively -0.674 and -0.817, P values below 0.01). In group H, there was obviously positive correlation between EVLWI and ITBVI (r = 0.751, P < 0.01) but no obvious correlation between EVLWI and PVPI, oxygenation index and EVLWI, or oxygenation index and PVPI (with r values respectively -0.275, 0.197, and 0:062, P values above 0.05). The total area under ROC curve of PVPI value for differentiating the type of pulmonary edema was 0.987 [with 95% confidence interval (CI) 0.962-1.013, P < 0.01], and 1.9 was the cutoff value with sensitivity of 94.1% and specificity of 95.2% . The total area under ROC curve of ITBVI value for differentiating the type of pulmonary edema was 0.940 (with 95% CI 0.860-1.020, P < 0.01), and 1,077. 5 mL/m2 was the cutoff value with sensitivity of 95.2% and specificity of 88.2%.

CONCLUSIONSEVLWI, PVPI, and ITBVI have an important significance in the differential diagnosis of the type of burn-induced pulmonary edema, and they may be helpful in the early diagnosis and management of burn-induced pulmonary edema.

Blood Gas Analysis ; Blood Volume ; Burns ; complications ; Capillary Permeability ; Diagnosis, Differential ; Extravascular Lung Water ; Humans ; Lung ; blood supply ; Lung Injury ; physiopathology ; therapy ; Monitoring, Physiologic ; Pulmonary Edema ; diagnosis ; etiology ; ROC Curve ; Respiration, Artificial ; Retrospective Studies

Ranpirnase (onconase, ONC) is a new drug, with weak RNase activity and strong cytotoxicity to various tumor cells in vitro and in vivo. This study is to obtain recombination onconase (rONC) with high bioactivity. Based on the codon preference of Pichia pastoris, we designed and synthesized the gene according to cDNA sequences of ONC and the α mating factor's prepeptide. We screened positive clones after transforming the recombination plasmids into P. pastoris X-33, GSS115 and SMD1168. We screened the best combination of seven different vectors and host strains. Moreover, we optimized culture condition in shake flasks and 10 L bioreactor, and purified rONC from the supernatant after inducing it with 0.25% methanol by aqueous two-phase extraction coupling G50 molecular exclusion method. The highest rONC production was 13 mg/L in pPICZα-A/X-33/ONC combination under the condition of pH 5.5 and 23 degrees C in shake flasks for 7 d; and that the highest rONC production was 180 mg/L when the induction is performed in the lower basic salt medium with pH 5.5 in the 10 L bioreactor for 7 d. The yield of rONC is more than 90% at a purity of above 95%. rONC can kill various tumor cells in vitro. The expression and purification of rONC would be useful for further investigation of this new drug.
Antineoplastic Agents ; metabolism ; Bioreactors ; Cell Line, Tumor ; Codon ; DNA, Complementary ; Genetic Vectors ; Humans ; Pichia ; metabolism ; Recombinant Proteins ; biosynthesis ; Ribonucleases ; biosynthesis

Objective To investigate the effect and regulatory mechanism of autophagy related multifunc-tional protein p62/SQSTM1 on biological behavior in non-small cell lung cancer(NSCLC).Methods RT-qPCR was used to detect the expression of p62 in normal lung cells and NSCLC cells.CCK-8,wound-healing and Transwell assays were used to detect the effects of inhibition and promotion of p62 expression on the proliferation,migration and invasion in NSCLC cells.Western blotting was used to detect the effects of inhibition and promotion of p62 expression on the expression of apoptosis-related proteins(Bcl-2 and Bax)and autophagy-related proteins(ATG5 and Becline1)in NSCLC cells.A nude mouse transplantation tumor experiment was used to detect the effect of inhibiting p62 expression on the tumor volume and mass of NSCLC cells in vivo.Results Compared with that in normal lung cells,the expression level of p62 in A549 cells was the highest.Cell function experiments in vitro showed that inhibition of p62 expression reduced the abilities of proliferation,migration and invasion in A549 cells,and suppressed autophagy and induced apoptosis.Consistently,p62 overexpression has the opposite effects.In addi-tion,animal experiments in vivo showed that inhibition of p62 expression decreased the tumor volume and mass of tumor-bearing mice.Conclusion p62 could promote the growth of NSCLC A549 cell in vivo and in vitro by modu-lating autophagy.

Objective: To study the antiseptic effect of compound lysostaphin disinfectant and its preventive effect on infection of artificial dermis after graft on full-thickness skin defect wound in rats. Methods: (1) Each one standard strain of Klebsiella pneumoniae, Acinetobacter baumannii, and Staphylococcus aureus were selected. Each 20 clinical strains of Klebsiella pneumoniae, Acinetobacter baumannii, and Staphylococcus aureus were collected from those isolated from wound exudates of burn patients hospitalized in our wards from January 2014 to December 2016 according to the random number table. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of compound lysostaphin disinfectant to above-mentioned strains were detected. The experiment was repeated 3 times. Compared with the corresponding standard strain, the clinical strain with higher MIC and/or MBC was considered as having decreased sensitivity to the disinfectant. The percentage of strains of each of the three kinds of bacteria with decreased sensitivity was calculated. (2) Artificial dermis pieces were soaked in compound lysostaphin disinfectant for 5 min, 1 h, 2 h, and 4 h, respectively, with 21 pieces at each time point. After standing for 0 (immediately), 12, 24, 36, 48, 60, 72 h (with 3 pieces at each time point), respectively, the diameters of their inhibition zones to standard strains of Klebsiella pneumoniae, Acinetobacter baumannii, and Staphylococcus aureus were measured. The experiment was repeated 3 times. The shortest soaking time corresponding to the longest standing time, after which the disinfectant-soaked artificial dermis could form an effective inhibition zone (with diameter more than 7 mm), was the sufficient soaking time of the disinfectant to the artificial dermis. (3) Forty Sprague-Dawley rats were divided into post injury day (PID) 3, 7, 14, and 21 sampling groups according to the random number table, with 10 rats in each group. A full-thickness skin defect wound with a diameter of 20 mm was made on both sides of the spine on the back of each rat. Immediately after injury, the artificial dermis without any treatment was grafted on the wound on left side of the spine (hereinafter referred to as control wound), while the sufficiently soaked artificial dermis with compound lysostaphin disinfectant was grafted on the wound on right side of the spine (hereinafter referred to as disinfectant wound). On PID 3, 7, 14, and 21, the gross condition of wounds of all the surviving rats was observed, and the new infection rates of control wounds and disinfectant wounds were calculated. Then, the rats in the sampling group with corresponding time were killed, and the full-thickness wound tissue containing artificial dermis was collected for quantitative analysis of bacteria. Bacteria content of the uninfected control wounds and that of the uninfected disinfectant wounds were compared. Data were processed with chi-square test and Wilcoxon rank sum test. Results: (1) The MIC of compound lysostaphin disinfectant to standard strains of Staphylococcus aureus, Klebsiella pneumoniae, and Acinetobacter baumannii were 1/32, 1/32, and 1/512 of the original concentration of the disinfectant, respectively, and the MBC were 1/32, 1/16, and 1/512 of the original concentration of the disinfectant, respectively. The percentages of clinical strains of Klebsiella pneumoniae, Acinetobacter baumannii and Staphylococcus aureus with decreased sensitivity to compound lysostaphin disinfectant were 15% (3/20), 20% (4/20), and 10% (2/20), respectively. (2) After being soaked in compound lysostaphin disinfectant for 2 and 4 h, the longest standing time, after which the artificial dermis could form an effective inhibition zone against Klebsiella pneumoniae, Acinetobacter baumannii, and Staphylococcus aureus, were 24, 36, and 48 h respectively, longer than 12, 24, and 24 h of soaking for 5 min and 24, 24, and 36 h of soaking for 1 h. The sufficient soaking time of compound lysostaphin disinfectant to artificial dermis was 2 h. (3) On PID 3, no infection symptom was observed in all the wounds, and so both the new infection rate of control wounds and that of disinfectant wounds were 0. The artificial dermis was transparent but not well connected with the wound. On PID 7, the new infection rate of control wounds was 20.00% (6/30), which was obviously higher than 3.33% (1/30) of disinfectant wounds, χ²=4.043, P<0.05. On the infected wound, a large amount of purulent exudates were observed, and the artificial dermis was not connected with the wound and degraded partially. On the uninfected wound, artificial dermis was transparent and had a partial connection with the wound. On PID 14 and 21, no new infected wound was observed, and so both the new infection rate of control wounds and that of disinfectant wounds were 0. There was no obvious improvement on the infected wounds. The collagen layers of artificial dermis in the uninfected wound established a good connection with the wound and were separating from the silica gel layer gradually. Infection occurred in 2, 3, 1 control wound (s) in PID 7, 14, and 21 sampling groups, respectively, and in 1 disinfectant wound in PID 14 sampling group. The bacteria content of the infected wounds tissue was 0.79×10⁶ to 7.22×10⁹ colony-forming unit (CFU)/g. The bacteria content of uninfected control wounds tissue in PID 3, 7, and 14 sampling groups were (3.43±1.88)×10^2, (2.37±0.43)×10^3, and (8.40±1.03)×10³ CFU/g, respectively, which were significantly higher than (0.33±0.12)×10^2, (0.43±0.17)×10^3, (2.16±0.52)×10³ CFU/g of uninfected disinfectant wounds tissue (Z=-3.780, -3.554, -3.334, P<0.05). The bacteria content of uninfected control wounds tissue and that of uninfected disinfectant wounds tissue in PID 21 sampling group were similar (Z=-0.490, P>0.05). Conclusions Compound lysostaphin disinfectant has quite strong antibacterial ability against Klebsiella pneumoniae, Acinetobacter baumannii, and Staphylococcus aureus. Clinical strains of the three kinds of bacteria were highly sensitive to compound lysostaphin disinfectant. Saturation of absorption of compound lysostaphin disinfectant achieves in artificial dermis after 2 hours′ soaking. After 24, 36, and 48 hours′ standing, the soaked artificial dermis still has the antibacterial effect on Klebsiella pneumoniae, Acinetobacter baumannii, and Staphylococcus aureus, respectively. The infection rate and the bacteria content of full-thickness skin defect wound in rats are all decreased when grafted with soaked artificial dermis.