OBJECTIVE: To investigate the curative effects of nanometer silver dressing and recombinant bovine basic fibroblast growth factor gel on burn residual wounds.METHODS: Forty burn patients with residual wounds because of deep second degree burn and full-thickness burn, were randomly divided into control group and management group. There were 20 patients in both groups. The patients of management group were treated by nanometer silver dressing and recombinant bovine basic fibroblast growth factor gel. The patients of control group were treated by saline and paraffin absorbent gauze. Healing time, wound healing rates at different time points,cases of infected wound and results of bacterial culture before and 7 days following treatment, and drug adverse reaction were recorded.RESULTS: The healing time of management group was significantly shorter than the control group (P < 0.01). The wound healing rates of management group was significantly higher than the control group at different time points (P< 0.01). The cases of infected wound was significantly fewer than the control group after treating (P < 0.01). The pathogenic bacteria detection rate was significantly lower than the control group after 7 days (P < 0.01).CONCLUSION: There was better antibacterial activity, decurtating the healing time when the management of nanometer silver dressing and recombinant bovine basic fibroblast growth factor gel on burn residual wounds were put into practice.

Objective To investigate the effect of different programs of preconditioning with hyperbaric oxygen (HBO) on spinal cord ischemia-reperfusion injury (I/R) in rabbits. Methods Forty-five New Zealand rabbits aged 4-5 months weighing 2.0-2.5 kg were randomly divided into 5 groups: group S, sham operation ( n = 5);group IR, spinal cord I/R injury (n = 10);group H_(1～3) , the animals were pretreated with 100% O_2 at 2.5 ATA 1 h/d for 5 (group H_1 ), 10 (group H_2 ) , or 20 (group H_3 ) consecutive days respectively 24 h before spinal cord I/R. The animals were anesthetized with iv pentobarbital sodium 30 mg/kg. The artery in the ear and left femoral artery were cannulated for proximal and distal mean blood pressure monitoring. Spinal cord ischemia was produced by cross-clamping of abdominal aorta distal to renal artery for 20 min. Hind-limb motor function was assessed at 48 h after reperfusion according to the modified criteria established by Tarlov (0 = no spontaneous movement, 4= normal motor function) . The animals were then killed and the L_5 segment of the spinal cord was removed for detection of neuronal survival (by HE staining), apoptosis (by TUNEL) and degeneration (by Fluoro-Jade B staining). Results Preconditioning with 5 or 10 d of HBO improved the hind-limb motor function and preserved more normal neurons in the spinal cord after I/R injury. Both apoptotic and degenerative cell death were attenuated in H_1 and H_2 groups. There was no significant difference in hind-limb motor dysfunction and the number of normal neurons in the lumbar spinal cord between H_3 group and I/R group. Conclusion Preconditioning with 5 d or 10 d HBO induces tolerance against spinal cord I/R injury, whereas preconditioning with 20 d of HBO fails to protect the spinal cord from I/R injury.

In this study, the relationship between mycelium morphology and laccase production was studied. The results indicated that the morphology of P. ferulae pellets was changed when glass beads were added. Laccase production showed higher with spherical mycelium than with filamentous or flocculent mycelium. In addition, the spherical mycelium with a diameter of 0.2-0.4 mm highly affected laccase production. Effect of the composition of culture medium on pellets was investigated and results indicated that various concentrations of glucose, corn meal and wheat bran were important to the formation of pellets in diameter of 0.2-0.4 mm. Besides nutrients, the addition of non-nutritional substrates influenced the distribution of P. ferulae pellets. However, the production of laccase was not promoted by non-nutritional substrates.
Culture Media ; Fermentation ; Glass ; chemistry ; Industrial Microbiology ; Laccase ; biosynthesis ; Mycelium ; cytology ; growth & development ; Pleurotus ; cytology ; enzymology

[ ABSTRACT] AIM: To investigate the effects of bone marrow mesenchymal stem cells ( BMSCs) modified by programed death ligand-1 immunoglobulin ( PDL1 Ig) gene on immune rejection of orthotopic liver transplantation in rats. METHODS:Rat BMSCs were cultured and identified.The protein expression of PDL1 Ig in the BMSCs 72 h after infection with pAdEasy-1/PDL1 Ig was detected by Western blot.Mixed lymphocyte reaction was used to detect the inhibitory effect of BMSCs on the viability of T-lymphocytes in peripheral blood.The male Wistar rats were used as donors (n=40), and the male SD rats were used as recipients ( n=40 ) .The rat model of orthotopic liver transplantation was established by im-proved cuff method for observing acute rejection.The rats were randomly divided into control group, BMSCs treatment group, BMSCs/GFP treatment group and BMSCs/PDL1 Ig treatment group with 10 pairs each.Five rats were executed at the 7th day and the remains were used for measuring the survival time.RESULTS:The expression of PDL1 Ig in the BM-SCs was detected after pAdEasy-1/PDL1 Ig infection.The effect of BMSCs/PDL1 Ig on the viability of the lymphocytes was stronger than that of BMSCs/GFP.The level of IL-4 in BMSCs/PDL1 Ig group was significantly higher than that in the other 3 groups, while the levels of IFN-γand IL-2 were significantly decreased.The liver function in BMSCs/PDL1 Ig group was significantly improved and the levels of ALT, AST and TBil were almost recovered to normal at the 7th day after transplan-
tation.Severe rejection reaction was observed in control group, and rejection reactions were decreased with different degrees in BMSCs treatment group and BMSCs/GFP treatment group.Much slighter rejection reaction and significantly longer sur-vival time were showed in BMSCs/PDL1 Ig group than those in the other 3 groups.CONCLUSION:PDL1 Ig-modified BM-SCs inhibit the rejection of liver transplantation in rats and induce the immune tolerance, and the effect is better than that of BMSCs alone.

Objective To investigate the changes of the expressions of heat shock proteins 70 (HSP70) and insulin and HSP70 protect effect on the gastric mucosa of rats with scald injury, and explore the relationship between insulin and HSP70 . Methods With a model of 30 total body surface area (TBSA) full-thickness burned rats,the expression and distribution of HSP70 in the gastric mucosa was detected with immunohistochemical method and analyzed by a micro-image analysis system, and at the same time the pathological changes of the gastric mucosa tissue of each group were analyzed by Western blot and immunohistochemistry at the 3rd,6th,12th,24th and 48th hour postburn. Results The expression of HSP70 obviously decreased at the 48th hour post scald injury. The expression of HSP70 in the treatment group was significantly higher than that in the scalded group at most time points except the 12th(9.40±1.52,P＝0.065) hour and at equal parallel time［(6.80±1.10,8.60±0.55,10.80±1.64,11.40±1.34),P＜0.05］. The gastric mucosal injury index in the scalded group was significantly higher than that in the treatment group and at equal parallel time ［(4.05±0.36,11.97±1.15,20.98±2.83,13.92±0.94,1.60±0.55),P＜0.05］. In pathological observation, the control group manifested the intact gastric mucosal tissue formation, the scalded group showed obvious gastric mucosal tissue injury in the early phase of scald injury, while the treatment group showed less severe injury than the scald group. A positive correlationwas found in the gastric mucosal injury index and HSP70(r＝0.904,P＜0.01) and also between the serum glucose and HSP70(r＝0.961,P＜0.01).Conclusions Insulin increased the expression of HSP70 and decreased the gastric mucosal injury index in the gastric mucosal tissue of SD rats in the phase of scald injury. It may be one of the vital mechanisms of insulin protecting the gastric mucosal tissue.

Objective To establish a flow cytometric measurement of detecting minimal residual disease(MRD) according to the leukemia-associated immunophenotypes in children with acute lymphoblastic leukemia(ALL) and to explore the significance of MRD detection in ALL children for a individualized treatment. Methods A variety of four-color fluorescent antibody combinations were used to investigate the children's normal bone marrow. The normal bone marrow pattern at two-parameter plots was established to identify the residual tumor cells, seventy-five bone marrow samples from newly diagnosed ALL children were analyzed with four-color cytometry to determined the optimal combinations which can clearly distinguish the tumor cells from normal cells. The bone marrow samples were monitored with the combination panel in 60 patients at the end of induction therapy and follow-up treatment. Cytomorphology test, PCR amplification of 29 fusion genes as well as IgG and TCR gene rearrangements were performed simultaneously. Results Sixty-nine cases (92.0%) could be identified for effective antibody combinations to monitor MRD by four-color cytometry. Fusion genes or IgG and T cell receptor (TCR) gene rearrangements can be detected in 21 cases (28.0%) to monitor MRD by PCR. No MRD can be detected in 25 bone marrow samples at the end of induction therapy and follow-up treatment. Four-color cytometry could detect as low as 0.021%-4.130% residual leukemia cells. Conclusion MRD can be monitored by flow cytometry which is faster than PCR, and the sensitivity is superior to morphology method.

This study was to evaluate the cellular inhibition effects induced by anti-AGR2 monoclonal antibody 18A4 in two AGR2-negative melanoma cells, A375 and B16-F10, treated with external oncoprotein AGR2. MTT assay and colony formation assay were performed to detect the cell proliferation rate. Flow cytometry analysis was performed to evaluate cell cycle transition. Cell migration rate was analyzed by wound healing assay. Cell morphological changes were detected by phalloidin staining. The expression of p53 was detected by Western blotting and immunofluorescence. Results showed that 18A4 inhibited the AGR2-dependent tumor properties including enhanced proliferation, accelerated cell cycle, increased cell migration and morphological changes of cells. In addition, by Western blot analysis and immunofluorescence, AGR2 blocking antibody 18A4 also restored p53 expression that was repressed by external AGR2 treated by chemotherapeutic drug doxorubicin. These findings suggest that 18A4 is able to inhibit the cellular tumorigenic properties induced by external AGR2 and is a potential drug against fumor.

Objective:To evaluate the efficacy of esketamine combined with different doses of remimazolam for induction of general anesthesia in pediatric patients.Methods:One hundred and sixty pediatric patients of either sex, aged 3-6 yr, of American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ, with body mass index of 13-20 kg/m 2, undergoing elective general anesthesia under a laryngeal mask, were divided into 4 groups ( n=40 each) by the random number table method: esketamine combined with propofol group (KP group) and esketamine combined with different doses of remimazolam group (0.2, 0.3, 0.4 mg/kg) groups (KR1 group, KR2 group, KR3 group). Esketamine 0.8 mg/kg was intravenously injected in the preanesthesia room. After entering the operating room, propofol 2.5 mg/kg was intravenously injected in KP group, and remimazolam 0.2, 0.3 and 0.4 mg/kg were intravenously injected in KR1, KR2 and KR3 groups, respectively. When the child lost consciousness and the Modified Observer′s Assessment of Alertness/Sedation Scale score<1, sufentanil and mevacurium were intravenously injected. When the Modified Observer′s Assessment of Alertness/Sedation Scale score≥1, rescue sedation was performed, and 3 min later the laryngeal mask airway was inserted. The onset time of sedation, response to laryngeal mask airway placement, rescue sedation, hypotension, tachycardia, bradycardia, bucking, hiccup, injection pain and apnea were recorded, and the increase rate of perfusion index (PI) was calculated. Results:No response to laryngeal mask implantation occurred in the four groups. Compared with KP group, the onset time of sedation was significantly prolonged, the incidence of hypotension, bradycardia, injection pain and apnea was decreased, the incidence of tachycardia was increased, and the increase rate of PI was decreased in KR1, KR2 and KR3 groups, and the rate of rescue sedation and incidence of bucking were increased in KR1 and KR2 groups ( P<0.05). Compared with KR1 group, the onset time of sedation was significantly shortened in KR2 group and KR3 group, and the rate of rescue sedation and incidence of bucking were decreased in KR3 group ( P<0.05). Compared with KR2 group, the onset time of sedation was significantly shortened, and the rate of rescue sedation was decreased in KR3 group ( P<0.05). There was no significant difference in the increase rate of PI, hypotension, bradycardia, tachycardia, injection pain and apnea among KR1, KR2 and KR3 groups ( P>0.05). There was no significant difference in the incidence of hiccup among the four groups ( P>0.05). Conclusions:Esketamine 0.8 mg/kg combined with remimazolam 0.4 mg/kg can be safely and effectively used for anesthesia induction and has milder inhibition of respiration and circulation as compared with esketamine combined with propofol in pediatric patients.

Objective: To evaluate an assay permitting amplification of target 5′-untranslated region (5′-UTR) sequences directly from clinical specimens and distinction among serotypes of enterovirus (EV). Methods: A total of 518 rectal swabs and 148 nasal swabs tested positive by pan-enterovirus real-time PCR were collected. 5′-UTR and the viral protein 1 (VP1) gene fragments were amplified and sequenced separately for serotyping. The inconsistent samples by 5′-UTR and VP1 serotyping were further determined by using the serotype-specific RT-PCR. Results: A total of 553 (83.0%) samples were detected by 5′-UTR serotyping and 318 (47.7%) were detected by VP1 serotyping in all 666 positive specimens, and there was significant difference in the detection rates between two methods in rectal and nasal swabs (P<0.001). For the rectal swabs, the mainly detected serotypes were CoxA6 (217), CoxA16 (88), EVA71 (40), CoxA10 (28) and CoxA4 (27) by 5′-UTR serotyping. Compared with the VP1 serotyping, the sensitivity and specificity of 5′-UTR serotyping were 57.1%-100% and 67.4%-98.1% respectively, with varied consistence with serotypes (kappa value 0.214-0.283). For the nasal swabs, the most frequently detected serotype was EVD68, with the sensitivity of 100%, the specificity of 91.1%, and the poor consistence (kappa value 0.217). CoxA6, CoxA16, EVA71, CoxA10 and EVD68 were further confirmed by serotype-specific RT-PCR. Using VP1 serotyping combined with serotype-specific RT-PCR as a reference method , the effect of performance of 5′-UTR serotyping on diagnosis was increased. Conclusions The performances of 5′-UTR serotyping in enterovirus vary with serotypes. The application of 5′-UTR serotyping should be considered comprehensively according to the purpose of the study.

To produce recombinant phospholipase A(1) (PLA(1)) by Escherichian coli, the pla gene encoding PLA(1) was amplified from Serratia liquefaciens by PCR and cloned into two vectors pET20-b(+) and pET28-a(+). The two recombinant plasmids were then transformed into E. coli BL21 (DE3) individually to express PLA(1). E. coli BL21(DE3)/pET28a-pla yielded extracellular PLA(1) with an activity of 40.8 U/mL in batch cultivations of shaken flasks by auto-induction, which was accounted for 91% of total enzyme activity. On the basis of primal auto-induction medium, the optimized fermentation medium of PLA(1) contained tryptone 10 g/L, yeast extract 5 g/L, glucose 0.8 g/L, lactose 5 g/L, Na2HPO4 25 mmol/L, KH2PO4 25 mmol/L and 1 mmol/L MgSO4 (final concentration). Glycine (7.5 g/L) was added 6 h after inoculated. After incubated at 37 degrees C for 24 h, extracellular enzyme activity reached 128.7 U/mL.
Cloning, Molecular ; Culture Media ; Escherichia coli ; genetics ; growth & development ; metabolism ; Fermentation ; Lactose ; pharmacology ; Phospholipases A1 ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Serratia liquefaciens ; enzymology