[Objective] To investigate the factors influencing the detection of HLA-C expression by flow cytometry. [Methods] A total of 12 hematopoietic stem cell suspension samples from peripheral hematopoietic stem cell volunteer donors were randomly collected after CD34⁺ cell counting detection. The influence of detecting different number of nucleated cell (500 000, 50 000 and 5 000), sequential order of red blood cell lysis and antibody incubation, and the HLA-C antibody with varied remaining time from the expiration date on the detection results of HLA-C expression by flow cytometry were investigated, respectively. The significance of differences between different groups was analyzed through Student t test. [Results] There was no significant difference in the proportion of HLA-C positive cells and mean fluorescence intensity (MFI) among the three groups with different nucleated cell numbers detected (500 000, 50 000 and 5 000) (P>0.05). The sequential order of red blood cell lysis and antibody incubation had no influence on the proportion of HLA-C positive cells (P>0.05), but HLA-C MFI value was significantly lower when antibody incubation was performed after red blood cell lysis than that when antibody incubation was performed before red blood cell lysis (P<0.05). The proportion of HLA-C positive cells and MFI value detected by HLA-C antibody remaining 24 months from the expiration date were significantly higher than those detected by HLA-C antibody remaining only 5 months from the expiration date (P<0.05). [Conclusion] The present study has investigated the factors of influencing HLA-C expression level by flow cytometry, the results have important reference and application value for standardizing the experimental operation of HLA-C expression and improving the accuracy and comparability of detection results.

Objective:To establish the prokaryotic expression system of Taenia solium (Ts) 14-3-3.2, and observe the expression of Ts14-3-3.2 protein at the stages of Ts adult and cysticercus. Methods:Based on the Ts14-3-3.2 gene sequence obtained by the Department of Parasitology, Zunyi Medical University in the previous study, the whole gene was synthesized by PCR-based accurate synthesis (PAS) method. After double digestion with restriction enzymes Nde Ⅰ and Xba Ⅰ, the plasmid pCzn1 was ligated to construct a recombinant plasmid pCzn1-Ts14-3-3.2. Then it was transformed into Escherichia coli ArcticExpress competent cells to induce the expression of Ts14-3-3.2 protein. The expression products were analyzed and identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and coomassie blue staining. The purified Ts14-3-3.2 recombinant protein was obtained by Ni-affinity chromatography. New Zealand rabbits were immunized with the recombinant protein to produce Ts14-3-3.2 polyclonal antibody. Western blotting was used to detect the expression of Ts14-3-3.2 protein at the stages of Ts adult and cysticercus. Results:The recombinant plasmid pCzn1-Ts14-3-3.2 was successfully constructed. After induced expression, Ts14-3-3.2 target protein bands appeared in the supernatant and precipitated at the relative molecular weight of about 29.31 × 10 3. The purified Ts14-3-3.2 recombinant protein with His label could be recognized by anti-His monoclonal antibody, and the Ts14-3-3.2 polyclonal antibody with titer of 1 ∶ 512 000 was obtained. Western blotting showed that Ts14-3-3.2 protein was expressed at the stages of Ts adult and cysticercus. Conclusions:The prokaryotic expression system of Ts14-3-3.2 is successfully established, and the Ts14-3-3.2 polyclonal antibody with relatively higher purity and titer is obtained. The Ts14-3-3.2 protein is expressed at the stages of Ts adult and cysticercus.

Parasitic diseases are diseases caused by parasites invading the human body, and 14-3-3 family plays an important role in the life activities of parasites, such as infection, growth and development, reproduction, and so on. Carrying out systematic research on it will help to understand the distribution and biological functions of the 14-3-3 family in parasites, and provide help for the immunological diagnosis and prevention of parasitic diseases. This article reviews the recent progress in the study of 14-3-3 family of important human parasites.

OBJECTIVETo establish a kidney deficient aging model (KDAM), assess it in antioxidant capacity, HPAT axis function and bone metabolism, and compare with D-galactose aging model.

METHODAging rat model was established by injecting D-galactose solution, meanwhile dexamethasone solution was injected to establish kidney deficient aging model. Then these models were evaluated by serum MDA (malondialdehyde) and GSH-Px (glutathione peroxidase), liver SOD (superoxide dismutase), adrenal, thymus and spleen index, CD4(+), CD8(+), and serum COR (cortisol), BGP (bone Gla-protein), plasma ACTH (adrenocorticotropic hormone) and CRH (corticotropin-releasing hormone).

RESULTCompared with the normal group, the aging model group and the kidney deficient aging group showed significant decrease in liver SOD activity (P < 0.01 on average) and significant increase in serum MDA content (P < 0.01 on average) , and the kidney deficient aging group revealed remarkable decline in plasma ACTH content (P < 0.05). Compared with the normal group and the aging model group, the kidney deficient aging model group's weight, serum GSH-Px decreased (P < 0.01, P < 0.05), adrenal index decreased (P < 0.05, P < 0.01), serum COR decreased (P < 0.05 on average), plasma CRH increased (P < 0.05, P < 0.01), serum BGP content significantly decreased (P < 0.01 on average), value of CD4(+), CD8(+) decreased (P < 0.05, P < 0.01), CD4(+)/CD8(+) increased, but without significant difference.

CONCLUSIONThe kidney deficient aging model shows significant decrease in antioxidant capacity, dysfunction of HPAT axis disorder and abnormal bone metabolism. However, D-galactose aging model only shows a significant difference in antioxidant capacity.

Aging ; drug effects ; physiology ; Animals ; Antioxidants ; metabolism ; Disease Models, Animal ; Galactose ; adverse effects ; Humans ; Kidney ; physiopathology ; Kidney Diseases ; chemically induced ; metabolism ; physiopathology ; Liver ; enzymology ; Male ; Malondialdehyde ; metabolism ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

OBJECTIVETo study the anti-inflammatory effect of the combination of Epimedii Folium, Ligustri Lucidi Fructus and dexamethasone on asthma and its effect in inhibiting adverse reaction against hormone.

METHODDexamethasone had been injected intraperitoneally into asthmatic model rats for 2 weeks, together with the oral administration of Epimedii Folium and Ligustri Lucidi Fructus. The contents of IL-4, INF-gamma, IL-5 and GCR in BALF, serum COR, plasma ACTH and hypothalamic CRH were observed.

RESULTThe combination of Epimedii Folium, Ligustri Lucidi Fructus and dexamethasone can significantly decrease the contents of IL-5 and IL-4 in BALF and ACTH and CRH content in plasma, increase the content of IFN-gamma and GCR in BALF and balance Th1/Th2.

CONCLUSIONThe combination of Epimedii Folium, Ligustri Lucidi Fructus and dexamethasone has a better anti-inflammatory effect on asthmatic model rats, and play a protective role in the pathway of endogenous dexamethasone.

Animals ; Asthma ; drug therapy ; immunology ; Bronchoalveolar Lavage Fluid ; immunology ; Cytokines ; analysis ; Dexamethasone ; administration & dosage ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; administration & dosage ; Ligustrum ; Male ; Rats ; Rats, Sprague-Dawley ; Th1 Cells ; immunology ; Th2 Cells ; immunology

Objective To evaluate the efficacy and safety of percutaneous nephrolithotomy (PCNL) under ultrasonic guidance for the treatment of upper urinary calculus. Methods From June 2004 to July 2009, 1289 patients with upper urinary tract calculi were treated with PCNL under ultrasonic guidance, 386 cases of them were upper ureteral calculi, 463 cases were single renal calculi,355 cases were multiple renal caculi, 85 cases were staghorn calculi. Calculus size was 0.8 cm× 1.2cm-9.0 cm× 5.3 cm,mean 2. 1 cm× 3.1 cm. Nine hunderd and thirty-five cases were with hydronephrosis. Results All patients were performed PCNL successfully, without conversion to open surgery or death. Mean operation time was 90(55-220)min, mean operative blood loss was 175(60-800)ml. Thirty-one cases were given blood transfusion during operation or post operation, 14 cases were given super-selective renal artery embolism for bleeding control. 1105 cases were treated with single tract, 108 cases with double tracts, 76 cases with three tracts. Calculus clearance rate of onestage PCNL was 85.03%(1096/1289);193 cases were found with residual calculi, 67 of them were given extracorporeal shock-wave lithotripsy, 126 of them were given two-stage PCNL, calculi were removed complelety in 85 cases. Total calculus clearance rate was 91.62%(1181/1289). Conclusions PCNL under ultrasonic guidance is an effective and safe method for the treatment of upper urinary calculus with advantages of accurate positioning, high calculus clearance rate, minimal trauma, and fewer complications. The key of successful PCNL is selection of patients, grasp of operation technique, and availability of other equipements.

OBJECTIVETo study the neuro-endocrine adjustment effects of Herba Epimedii and Fructus Ligustrilucidi on the asthmatic rats.

METHODRat asthma model was duplicated by OVA (ovalbumin) through sensitizing and challenging. Fifty male rats were randomly divided into normal group, model group, adjustment group of Herba Epimedii and Fructus Ligustrilucidi, Peibenfang group and Asimei capsule group. Investigating levels of ET (Endothelin), NO, iNOS (inducible NOS ), and cNOS (constitutive NOS) in blood serum and BALF (bronchoalveolar lavage fluid ), CORT (corticotrophin) in serum, ACTH (adrenocorticotropin hormone) in plasma, CHR (corticotropin release hormone) in hypothalamus, protein expression of GCR (glucocorticoid receptor) in lung tissue.

RESULTThe adjustment of Herba Epimedii and Fructus Ligustrilucidi could inhibit ET and NO content in BALF (all P < 0.05), decrease the level of iNOS in serum and BALF (P < 0.01 or P < 0.05), and increase the level of cNOS in serum and BALF (P < 0.01 or P < 0.05), raise the concentration of serum CORT (P < 0.01), enhance the protein expression of GCR in lung tissue (P < 0.05).

CONCLUSIONThe preventive and therapeutic effect of Herba Epimedii and Fructus Ligustrilucidi on asthma relates to their adjustment effect on ET/NO and HPA axis.

Animals ; Asthma ; blood ; drug therapy ; metabolism ; Bronchoalveolar Lavage Fluid ; chemistry ; Disease Models, Animal ; Endothelins ; blood ; Humans ; Hypothalamo-Hypophyseal System ; Lung ; metabolism ; Male ; Nitric Oxide ; blood ; Nitric Oxide Synthase Type II ; blood ; Pituitary-Adrenal System ; drug effects ; metabolism ; Plant Extracts ; administration & dosage ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Glucocorticoid ; metabolism

To observe the effects of dexamethasone (DXM) on changes of syndrome and hypothalamic-pituitary-adrenal (HPA) axis of asthmatic rats at different stages of intervention (before withdrawal, during withdrawal and after withdrawal).

Objective To compare the safety of laparoscopic live donor nephreetomy(LDN) and open live donor nephrectomy(ODN), evaluate the kidney function and blood pressure of living donors during 1 year follow-up. Methods Thirty cases of LDN and 30 eases of ODN were retrospectively reviewed. The operation time, warm ischemia time, operative blood loss, time to post-operative intake and time to ambulation of the 2 grouups were compared. According to the modified Clavien classifica-tion system procedure-related complications were described and compared. Serum creatinine(SCr) le-vels, blood pressure and 24-h urine protein excretion were measured before nephreetomy and 1 d, 7 d, 3 months, 6 and 12 months after nephrectomy. Glomerular filtration rate (GFR) were measured preo-pratively and at 6 and 12 months postoperatively. These data were statistically analyzed. Results The operation time was (98. 6+13. 6)rain and (96.3+19. 5)rain in the LDN and ODN groups, re- spectively. Warm ischemia time in the LDN group was (90.6±15.1)s, in the ODN group was (86.4±12.3)s. Operative blood loss was (105.2±634.8)ml and (206.3±126.4)ml in the LDN and ODN groups(P<0.01). For the time to post-operative intake and time to ambulation, LDN group was (28.5±2.9)h and (25.8±63.8)h, ODN group was (38.6±63.3)h and (36.5±65.3)h(P<0.01). Perioperative complications rates were 6.6%(2/30) and 23.3%(7/30) for LDN and ODN, respective-ly. SCr was (109.1±7.5), (105.4±69.5), (96.6±10.7), (89.4±11.5), (91.6±69.3)/zmol/L in the LDN group and (107.3±69.6), (103.3±68.4), (95.4±69.1), (90.5±13.6), (90.3±11.7)μmol/L in the ODN group 1 day, 7 days, 3 months, 6 months and 12 months after nephrectomy. The mean GFR of LDN and ODN was 64.7 and 65.8 ml/min at 6 months after nephrectomy, 65.9 and 67.5 ml/min at 12 months postoperatively, which were significantly different comparing with preoperative mean GFR in each group(P<0.05) but no significant difference was found between 6 months and 12 months after nephrectomy and between the 2 groups at the same time point respectively(P>0.05). Mean 24 h protein excretion was elevated after either LDN or ODN during 1 year followup, but was not significantly different either between predonation and 1 year after nephrectomy or between the 2 groups at the same period. Blood pressure increased or decreased slightly with the duration of follow-up,no significant blood presure changes were found before and after nephrectomy or between the two groups at the same period postoperatively. Conclusions LDN has the advantages of minimal trauma, less operative blood loss and quicker convalescence. It is safe and and has no adverse effects regarding kidney function and blood pressure during the first year after living kidney donation comparing to ODN.