OBJECTIVE To optimize the molding process of Xiebai Granules(XG)using the Box-Behnken design-response surface method combined with the BP neural network method,and to evaluate the consistency of particle quality between different bat-ches by establishing physical fingerprint.METHODS Dry paste powder was used as the main drug,dry granulation was adopted,and the forming rate,dissolution rate,moisture absorption rate and angle of repose of the granules were used as evaluation indexes,the ex-cipients dextrin,maltodextrin and lactose of the particles,were screened by single factor test combined with simplex-lattice design and entropy weight method,and the optimal excipient ratio was selected.The entropy weight method combined with the Box-Behnken de-sign-response surface method and the BP neural network algorithm were used to optimize the process parameters,and the process veri-fication was carried out.The physical fingerprint was used to comprehensively characterize the bulk density(Da),hygroscopicity(H),moisture(HR),tap density(Dc),angle of repose(α),Hausner ratio(IH),relative uniformity index(Iθ),Carr index(IC),and in-terparticle pore number(Ie),and the consistency of particle quality in different batches was evaluated.RESULTS The optimal ratio of excipients was dextrin 15%,maltodextrin 48%,and lactose 37%.The optimal process parameters were conveying speed 95 r·min-1,pressure wheel speed 4 r·min-1 and hydraulic pressure 7 MPa.The similarity of the physical fingerprints of the five bat-ches of XG was greater than 0.98.CONCLUSION The optimized molding process of XG is stable and feasible,and the quality of different batches of XG is stable,which can provide a reference for the development and industrial scale-up production of XG.

OBJECTIVE To optimize the molding process of Xiebai Granules(XG)using the Box-Behnken design-response surface method combined with the BP neural network method,and to evaluate the consistency of particle quality between different bat-ches by establishing physical fingerprint.METHODS Dry paste powder was used as the main drug,dry granulation was adopted,and the forming rate,dissolution rate,moisture absorption rate and angle of repose of the granules were used as evaluation indexes,the ex-cipients dextrin,maltodextrin and lactose of the particles,were screened by single factor test combined with simplex-lattice design and entropy weight method,and the optimal excipient ratio was selected.The entropy weight method combined with the Box-Behnken de-sign-response surface method and the BP neural network algorithm were used to optimize the process parameters,and the process veri-fication was carried out.The physical fingerprint was used to comprehensively characterize the bulk density(Da),hygroscopicity(H),moisture(HR),tap density(Dc),angle of repose(α),Hausner ratio(IH),relative uniformity index(Iθ),Carr index(IC),and in-terparticle pore number(Ie),and the consistency of particle quality in different batches was evaluated.RESULTS The optimal ratio of excipients was dextrin 15%,maltodextrin 48%,and lactose 37%.The optimal process parameters were conveying speed 95 r·min-1,pressure wheel speed 4 r·min-1 and hydraulic pressure 7 MPa.The similarity of the physical fingerprints of the five bat-ches of XG was greater than 0.98.CONCLUSION The optimized molding process of XG is stable and feasible,and the quality of different batches of XG is stable,which can provide a reference for the development and industrial scale-up production of XG.

Objective:To study the effects of trioxygen pretreatment on cerebral ischemia/reperfusion (I/R) injury in rats.Methods:A total of 24 clean grade male Sprague-Dawley (SD) rats were randomly divided into Sham group, brain I/R group (I/R group) and Ozone pretreatment group (Ozone group), with 8 rats in each group. The animals were routinely fed, and the operation was performed 5 days after the intervention of Ozone group by intraperitoneal injection of trioxygen water (concentration 80 mg/L, 0.01 mL/g), and the Sham group and I/R group were injected with equal volume normal saline. The Sham group only separated the arteries without ligation, and the I/R group and Ozone group established the rat cerebral I/R model. Neurological deficit score (NDS) was performed 2 hours after ischemia and modified neurological deficit score (mNSS) was performed 24 hours after reperfusion. Brain tissue was collected after anesthesia. Cerebral infarction was observed by 2, 3, 5-triphenyltetrazolium chloride (TTC) staining and the percentage of cerebral infarction volume was calculated. Protein expression of metabolic glutamate receptor 5 (mGluR5) and ionic glutamate α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptor (AMPAR) subunit GluA2 in cerebral ischemic penumbra was determined by Western blotting.Results:Compared with the Sham group, NDS score, mNSS score and percentage of cerebral infarction volume in I/R group were increased [NDS score: 2.63±0.52 vs. 0, mNSS score: 9.63±1.19 vs. 1.13±0.64, cerebral infarction volume: (41.25±2.93)% vs. 0%, all P < 0.05], and expressions of mGluR5 and GluA2 in penumbra area of cerebral ischemia were decreased [mGluR5 protein (mGluR5/β-actin): 0.44±0.14 vs. 1.00±0.10, GluA2 protein (GluA2/β-actin): 0.23±0.08 vs. 1.00±0.25, both P < 0.05]. Compared with the I/R group, mNSS score and percentage of cerebral infarction volume in the Ozone group were decreased [mNSS score: 7.00±1.20 vs. 9.63±1.19, cerebral infarction volume: (27.23±6.21)% vs. (41.25±2.93)%, both P < 0.05], and mGluR5 and GluA2 expressions in the penumbra of cerebral ischemia were up-regulated [mGluR5 protein (mGluR5/β-actin): 0.81±0.10 vs. 0.44±0.14, GluA2 protein (GluA2/β-actin): 0.76±0.13 vs. 0.23±0.08, both P < 0.05]. Conclusion:Trioxygen preconditioning can alleviate cerebral I/R injury in rats, and its mechanism may be related to the upregulation of GluR5 and GluA2 in the ischemic penumbra.

Objective To evaluate the effect of clemastine fumarate on Toll-like receptor 4/phosphatidylinositol-3-kinase/serine-threonine kinase (TLR4/PI3K/Akt) signaling pathway during hypoxia-reoxygenation (H/R) in rat cardiomyocytes.Methods H9C2 cells of rats cultured in vitro were seeded in culture wells or dishes at a density of 1×105 cells/ml and divided into 3 groups (n=11 each) by using a random number table method:control group (group C),H/R group and clemastine fumarate group (CF group).Cardiomyocytes were exposed to 5％ CO2-95％ N2in a low-glucose DMEM medium at 37℃ for 4 h followed by 4 h reoxygenation.At 4 h of reoxygenation,the cell viability was detected by CCK-8 assay,the ultrastructure was observed with a transmission electron microscope,the expression of TLR4,PI3K,phosphorylated Akt (p-Akt) and caspase-3 was detected by Western blot,and the expression of TLR4,PI3K and caspase-3 was detected by immunofluorescence.Results Compared with group C,the cell viability was significantly decreased,the expression of TLR4 and caspase-3 was up-regulated,and the expression of PI3K and p-Akt was down-regulated in group H/R (P＜0.05).Compared with group H/R,the cell viability was significantly increased,the expression of TLR4 and caspase-3 was down-regulated,the expression of PI3K and p-Akt was up-regulated (P＜0.05),and the mitochondrial damage was significantly attenuated in group CF.Conclusion The mechanism by which clemastine fumarate alleviates H/R injury to rat cardiomyocytes may be related to inhibiting TLR4 expression and activating PI3K/Akt signaling pathway.

Objective To investigate the effect of M2 macrophages resident in gastric cancer microenvironment on the tumor-promoting effect of human gastric cancer-derived mesenchymal stem cells (GC-MSCs).Methods Macrophages in BALB/c mice were depleted by using clodronate liposomes.The tumor volumes and weights in nude mice co-injected with GC-MSCs and BGC-823 with and without macrophage depletion were recorded.Tumor tissues of nude mice and gastric cancer patients were collected,and M2 macrophage-associated genes and proteins were detected by RT-PCR and western blot.Furthermore,the regulating effect of GC-MSCs on macrophage polarization to M2-subtype was validated in the co-culture experiment in vitro.Results Tumor growth in GC-MSCs co-injected mice was significantly inhibited by macrophage depletion (P=0.009).Results of RT-PCR and western blot showed that the transcription and expression of M2 macrophage-associated proteins were significantly higher in tumor tissues from GC-MSCs co-injected mice than those in the control group.Moreover,the transcription and expression levels of M2 macrophage-associated proteins were also high-er in gastric cancer tissues than those in the corresponding adjacent normal tissues.After co-culture with GC-MSCs directly,the expressions of M2 macrophage-associated proteins were significantly up-regulated in THP-1-derived macrophages.Conclusion M2 macrophages in gastric cancer microenvironment might play a critical role in the tumor-promoting effect of GC-MSCs.

Objective To evaluate the effect of sevoflurane preconditioning on the expression of metabotropic glutamate receptor type Ⅱ( mGluRⅡ) during focal cerebral ischemia-reperfusion ( I∕R) in rats. Methods Forty-eight clean-grade healthy male Sprague-Dawley rats were divided into 3 groups (n＝16 each) using a random number table method: sham operation group ( group S), cerebral I∕R group (group I∕R) and sevoflurane preconditioning group (group Sev). Rats were anesthetized with 10% chloral hydrate 3 ml∕kg. Focal cerebral I∕R was produced by occlusion of the right middle cerebral artery for 2 h fol-lowed by 24 h reperfusion. In group Sev, 2. 7% sevoflurane was inhaled for 1 h and 24 h later focal cerebral I∕R was produced. At 24 h after reperfusion, neurological deficit was scored, the cerebral infarct size was determined by TTC staining, the cell apoptosis in ischemic penumbra was observed by TUNEL, IκB-α ex-pression was detected by Western blot, and mGluRⅡexpression was determined by immunofluorescent stai-ning. The apoptosis rate was calculated. Results Compared with group S, the neurological deficit score, cerebral infarct size and apoptosis rate were significantly increased, the expression of mGluRⅡwas up-regu-lated, and the expression of IκB-α was down-regulated in I∕R and Sev groups ( P＜0. 05). Compared with group I∕R, the neurological deficit score, cerebral infarct size and apoptosis rate were significantly de-creased, the expression of mGluRⅡwas down-regulated, and the expression of IκB-α was up-regulated in group Sev (P＜0. 05). Conclusion Sevoflurane preconditioning reduces focal cerebral I∕R injury through inhibiting the expression of mGluRⅡ in rats.

Objective To detect the protein expression changes of mitochondrial apoptotic pathway during the reverse effects of lipid emulsion on bupivacaine cardiotoxicity, so as to investigate the probable mechanism concerning the reverse effect of lipid emulsion on bupivacaine cardiotoxicity.Methods The ventricular muscles of 15 healthy SD neonatal mice (1-3 d) were chosen to conduct primary culture in vitro.And the cardiomyocytes were cultivated in a medium containing bupivacaine for 24 hours to establish its bupivacaine poisoning model.The cultured cardiomyocytes were divided into three groups: control group (group C);bupivacaine group (group B);and bupivacaine+lipid emulsion group (group BL).Flow cytometry was applied to examine the apoptosis of cardiomyocytes, and the Western blot was employed to detect the protein expression variation of cytochrome C (Cyto-C) and cleaved casepase-3.Results Compared with group C, the apoptosis rate was remarkably increased in both group B and group BL and that of the group B was dramatically higher than that of the group BL, with a statistical significance (P<0.05).Compared with group C, the protein expression levels of both Cyto-C and cleaved casepase-3 were significantly increased in groups B and BL (P<0.05), and the protein expression levels of both Cyto-C and cleaved casepase-3 in group B were significantly higher than those in group BL (P<0.05).Conclusion Lipid emulsion can regulate apoptosis through inhibiting the release of mitochondrial Cyto-C and reducing casepase-3 activation, thus it protects cardiomyocytes.

Under the condition of information society, we should cultivate students'!ability of infor-mation literacy and innovation. Based on the early investigation of medical students'!information ability and scientific research quality, we carried on a staged and systematic subject-oriented information literacy training for post-graduate students in specialty of anesthesiology through such measures as enriching infor-mation resources, strengthening students'!awareness of information, the implementation of the supervisor group system or double tutor system, and other measures, which strengthened the graduate students' ! independent study ability and scientific research quality. The related practice can provide reference for the development of information literacy education in colleges and universities.

One of the major responsibilities for medical students is to save lives and to popularize emergency medical treatment knowledge for the masses of society.Based on research analysis,systemic training of cardiopulmonary resuscitation program including lectures,simulation education and social practice were performed for medical students.After practice activities,these students could proficiently apply the basic knowledge and skills of cardiopulmonary resuscitation in practical work and spread what they have learned to the public.Furthermore the program could improve students' abilities in social practice and team spirit and it is of great social significance.

In order to explore a new special and effective way to prevent graft versus host disease (GVHD) after allogenic bone marrow transplantation (allo-BMT), the stem cell antigen-1 (Sca-1) + early hematopoietic cells (EHC) from BALB/c mouse (H-2d) were introduced with exogenous mouse Fas ligand (mFasL) cDNA gene by the retrovirus-mediated gene transfer and expanded for one week, and then they were co-cultured with the spleen mononuclear cells (SMNC) from BAC mouse (H-2dxb) as one way mixed lymphocyte reaction (OWMLR). The cytotoxicity of treated BAC mouse SMNC against Na2 51CrO4 labeling SMNC from BALB/c mouse was observed. The bone marrow mononuclear cells (BMMNC) from BAC mouse treated by the above methods were transplanted into lethally-irradiated congenic BALB/c mice to observe the occurrence of GVHD. The results showed that the SMNC from BAC mouse after OWMLR with exogenous mFasL cDNA gene-transduced hematopoietic cells (HC) from BALB/c mouse in a ratio of 1 to 5 exhibited an obvious inhibition of the cytotoxicity against the BALB/c mouse spleen cells at different effector/target ratios as compared to the control group (P<0.01). The grade I GVHD or no GVHD and the 80% survival rate at day 60 post-BMT were observed in the BALB/c mouse receiving BAC mouse BMMNC treated with similar way, while the grade II - III GVHD and the 20% survival rate were noted in the control group (P<0.01). It is suggested that the attenuation of GVHD in allo-BMT recipient could be successfully achieved through FasL-Fas pathway in an H-2 haplotype disparate mouse combination.
Animals ; Bone Marrow Transplantation ; Fas Ligand Protein ; Female ; Graft vs Host Disease ; immunology ; therapy ; H-2 Antigens ; genetics ; Haplotypes ; Hematopoietic Stem Cells ; cytology ; immunology ; Membrane Glycoproteins ; immunology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Rats ; Rats, Wistar ; Signal Transduction ; Spleen ; cytology ; immunology ; T-Lymphocytes ; immunology ; Transfection ; fas Receptor ; immunology