Objective To examine the pathological changes in the liver of an AIDS patient with complicated infection of Pneumocystis carinii(PC). \ Methods\ A liver biopsy was made. The tissue was stained with HE, PAS, Giemsa, GMS, and acid\|fast staining, and examined under light microscope and transmission electron microscope. \ Results\ Granulomas (acid\|fast negative) in the tissue and numerous pathogens (PAS positive) in hepatic sinusoids were detected. Giemsa and GMS staining and electron microscopy all confirmed that the pathogen was Pneumocystis carinii. \ Conclusion\ The pathological findings revealed a diffuse extrapulmonary infection of Pneumocystis carinii in the patient of AIDS.

The successful establishment of animal models of cardiac arrest-cardiopulmonary resuscitation (CA-CPR) undoubtedly provided an important basis for exploring the method of cardiopulmonary resuscitation (CPR) and advanced cardiovascular life support (ACLS). However, pathophysiology varied with the etiology of cardiac arrest (CA). Therefore, preparation of similar animal models according to etiology was the basis for pathophysiological changes research. Compared with other animals, the rabbits had both the advantages of large and small animals, so they became common research object for the CA-CPR model. This paper reviewed the common methods of animal models of CA-CPR in rabbits. In this review, the methods, criteria, advantages, disadvantages and precautions of each model were analyzed, which would provide useful reference for CPR researchers.

Humanized monoclonal antibodies(mAbs) are increasingly widely used in targeted therapy for cancer and some other major diseases.Complementarity-determining region(CDR) grafting makes quantities of humanized mAbs available.Herein,we provide an overview on the strategy and progress of CDR grafting.

Objective To evaluate the treatment of current status and prognosis in childhood APL with APL2008 ,which was administrated since 2008 in our center .Methods A total of 43 children with newly diagnosed APL between 2008 to 2014 were studied retrospectively .Treatment options and current status were summarized from 28 patients who received APL2008 therapy . Results Studied 43 patients were at median age of 8 years and 4 months ,with 28 boys and 15 girls .The main clinical manifestations were infection ,anemia ,bleeding ,fever ,hepatomegaly ,splenomegaly and lymphadenopathy .The proportions of low ,intermediate and high risk groups were 27 .9% ,48 .8% and 23 .3% ,respectively .Eleven cases could be diagnosed as DIC .Bone marrow morphology showed abnormal elevation of promyelocyte .37 patients had distinctive immunophenotype such as frequent expression of CD33 , CD117 and MPO .PML/RARαfusion gene positive rate was 100% in 43 children and cytogenetic analysis were positive in 37 cases , of which specific genetic lesion in APL cells with t (15 ;17)(q22 ;q12) was found in 28 cases ,and karyotypes was found in 9 cases as infrequent chromosomal abnormalities .In 43 patients ,4 cases were early dead from intracranial hemorrhage at early stage ,and 11 cases were given up early .There were only 2 cases dead ,2 cases relapsed and 1 case lost among 28 APL children ,which enabled ef‐ficacy analysis possible .96 .4% of these 28 cases achieved HCR .The 2 year Kaplan Meier estimates of OS and EFS were 85 .9% ± 7 .6% and 80 .4% ± 8 .8% .But OS and EFS would be 94 .7% ± 5 .1% and 88 .9% ± 7 .4% if 3 patients who had non standard treat‐ment were excluded .Conclusion Childhood APL were characterized by anemia ,bleeding ,fever and infiltration .APL′s coincidence rate between PML/RARa fusion gene and morphology ,immunology and cytogenetics were 95 .3% ,90 .2% and 86 .5% ,respective‐ly .APL2008 significantly improved the prognosis of APL .

Objective To study the changes of PI3K/Akt/GSK3β signaling pathway during resuscitation with neck cooling in order to explore the relationship between the protective effect of neck cooling and the phosphorylation of PI3K/Akt and GSK3β.Methods Thirty rabbits were randomly(random number) divided into five groups, and models of cadiac arrest were induced by ventricular fibrillation(VF, the positive electrode in the right ventricle and negative pole on the apex of heart) for 4 min.In sham group,a electrode was placed into right ventricle without electric current conducted, and CA was not induced.The rabbits were sacrificed and specimens were taken at 24 hours after modeling.In normothermia treat group(NT group),resuscitation was carried out to restoration of spontaneous circulation(ROSC),and the rabbits were sacrificed and specimens were taken at 24 hours after modeling.In intra-arrest therapeutic hypothermia group (IATH group), rapid neck cooling was initiated at the same time with CPR,and the target brain temperature was set at 34 ℃ maintained for 4 hours after ROSC.Rabbits were sacrificed and specimens were taken at 24 hours after modeling.In recovery period cooling + LY294002 group(PATH+LY294002 group), LY294002 was injected intra-ventricularly at 20 minutes before resuscitation.Rapid neck cooling was started at the same time with CPR,and the target brain temperature was set at 34 ℃ maintained for 4 hours after ROSC.The rabbits were sacrificed and specimens were taken at 24 hours after modeling.In post-arrest therapeutic hypothermia group (PATH group), rapid neck cooling was begun after CPR for 1 hour,and the target brain temperature was set at 34 ℃ maintained for 4 hours after ROSC.The rabbits were sacrificed and specimens were taken at 24 hours after modeling.Animals were sacrificed by using overdose anesthetic drug.Western blot was used to detect the level of Akt p-Akt GSK-3β p-GSK-3β (ser9) protein, and TUNEL was used to observe apoptosis of tissues in each group.Multiple comparisons were performed with one-way analysis of variance (ANOVA).Results Compared with Sham group, Akt (Thr-308) phosphorylation (P-AKT) and P-GSK-3β levels in the brain neuron cytoplasm in 24 hours after CPR resuscitation in NT group was significantly reduced, and showed a gradual reduction trend (P<0.05);the P-AKT and P-GSK-3β levels in the brain neuron cytoplasm in 24 hours after CPR resuscitation in IATH group were significantly enhanced compared with NT group (P<0.05);the levels of these two kinds of protein at one hour after resuscitation in PATH group were significantly enhanced compared with NT group (P<0.05), but lower in IATH group.Intra-ventricularly injection of LY294002 made the effect of hypothermia lost, indicating that LY294002 inhibited the phosphorylation of Akt.Apoptosis cells were significantly reduced in IATH group and normothermia theatment group compared with PATH group and LY294002 group(P<0.05).Conclusions Neck cooling can reduce apoptosis in rabbit brain cells after recovery, and the protective effect on brain is best in intra-arrest therapeutic hypothermia group.LY294002 specifically block the PI3K/Akt pathway, and the protective effect of cooling on the brain can be abolished,indicating hypothermia protects the neurological function via activation of PI3K/Akt pathway.Neck cooling protects the neurological function by activating PI3K/Akt/GSK-3β, promoting the Akt activation, and increasing the expression of P-GSK3β.Specific Akt inhibitor LY294002 inhibits Akt phosphorylation of brain tissue recovery and further inhibit the phosphorylation of GSK-3β, thus abolishing protective effect of cooling on neurological function.

Objective To investigate the protectve effects and underlying mechanisms of deferroxamine on glutamate-induced injury in cultured hippocampal neurons.Methods Primarily cultured hippocampal neurons from fetal rat were used in a model of glutamate induced neurotoxicity.There were two experimental groups.Neurons were pretreated with deferroxamine before glutamate in the deferroxamine group, and neurons were treated with glutamate only in the control group.The morphological change was examined under microscope.Hoechst 33342 DNA staining method was used to study the ratio of condensed nuclei.The levels of lactate dehydrogenase (LDH), malonaldehyde (MDA) and hydroxyl radical were determined using biochemistry.The change in calcium signal was detected using microfluorescent technique.Results The neurons pretreated by deferroxamine had intact morphology with the ratio of condensed nuclei at 14% ± 6% compared to 58% ± 6% (t= 8.98, P ＜0.01 ) in the control group.LDH level was (36.42 ± 8.99) U/L in the deferroxamine group and was (68.06 ± 11.26) U/L in the control group ( t =3.25,P＜0.05).The respective levels of hydroxyl radical were (34.21 ±4.23) U/L and (47.06 ±8.79) U/L (t = 3.11, P ＜0.05 ).The respective levels of MDA were (12.26 ± 2.78 ) nmol/mg and (28.86±5.19) nmol/mg(t =4.88,P＜0.01).Conclusion Deferroxamine can protect neurons from glutamate induced damage.The mechanisms include an inhibition of Ca2+ overload and reduction in the levels of MDA and hydroxyl radicals.

Bacillus anthracis collagen-like protein(BclA) is a structural component of the exosporium filaments,as well as the immunodominant antigen on the spore surface.The genes encoding BclA proteins were cloned and sequenced from three Bacillus anthracis strains separated from China.It was founded that the BclA proteins of strain A16R and 40048,containing 388 and 322 amino acids,72 and 50 copies of GXX repeat,5 and 3 copies of 21-amino-acid sequence(GPT)_(5)GDTGTT(BclA repeat) respectively,are different from those reported by foreign scholars;while the BclA protein of strain 40022,containing 370 amino acids,66 copies of GXX repeat,and 5 copies of BclA repeat,is identical with that of strain 53169 reported by others.The results are helpful for the molecular typing of B.anthracis strains,and provide a basis for the elucidation of the pathogenesis and immunogenicity of B.anthracis spore.

OBJECTIVENeonatal hypoxic-ischemic encephalopathy (HIE) harms the lives and health of newborn infants and children severely. The prognosis is not satisfied, especially of the severe HIE. Mesenchymal stem cells (MSCs) can secrete a series of growth factors and neurotrophic factors. As well they have the potential ability to differentiate to the neural cells in vitro and in vivo. Therefore MSCs transplantation has been employed as a source of progenitor cells for cell therapy in patients with HIE in order to promote recovery of brain function and reduce the sequelae. Studies have shown that MSCs could enter the cerebral parenchyma and differentiate to neural cells through systemic infusion, but most of the researches applied adult stroke animal models. This study used neonatal HIE models to test the hypothesis that MSCs could enter the brain of newborn Wistar rats through the blood-brain barrier (BBB) by intraperitoneal infusion followed by observing the characteristics of the distribution and differentiation of MSCs in brain tissues, and exploring the effects of hypoxic-ischemic brain damage to the penetration and differentiation of MSCs.

METHODSIsolation and purification of MSCs were established from the whole bone marrow of juvenile Wistar rats by removing the nonadherent cells in primary and passage cultures. For cellular identification, MSCs of three to five passages were continuously pre-labeled with 5-bromo-2-deoxyuridine (BrdU) for 72 hours before transplantation. Animal models of HIE were built in 7-day-postnatal Wistar rats according to the method described by Rice. Two hours after hypoxia-ischemia, rats in HIE group (n = 8) were intraperitoneally infused with MSCs (4 x 10(6), 0.5 ml). In control group (n = 8), 7-day-postnatal normal Wistar rats were intraperitoneally infused with the same amount of MSCs. All rats were sacrificed and their cerebra were sectioned by cryomicrotome 14 days after transplantation. Immunohistochemical staining with chromogen diaminobenzidine (DAB) was used to detect and measure the cells derived from MSCs, and study the characteristics of distribution. To determine the differentiation of the BrdU positive cells entering the brains, immunofluorescence double labeling for BrdU and neural cells specific antigens was performed.

RESULTSMSCs were distributed throughout the cerebra in both groups at the 14th day after transplantation. The number of MSCs detected was 2415 +/- 226 in the control group, and 3626 +/- 461 in HIE group, respectively (t = 6.68, P < 0.05). More BrdU reactive cells were observed in the right ischemic hemisphere (1904 +/- 267) than in the contralateral hemisphere (1723 +/- 204), (t = 4.47, P < 0.05). No significant difference was found while comparing both cerebral hemispheres of the control group (t = 0.31, P > 0.05). In the HIE group, MSCs distributed more extensively, and some focal aggregations of MSCs were noticed. A few MSCs expressed Nestin-protein marker of neural progenitor cells, and almost none of the MSCs which expressed proteins characteristic of neuron (e.g. NSE) and astrocyte (e.g. GFAP) was detected at the 14th day after transplantation.

CONCLUSION1. MSCs could enter the cerebral parenchyma through BBB and migrate throughout the brain by intraperitoneal infusion. 2. More MSCs injected intraperitoneally were localized and directed to the sites of hypoxic-ischemic brain damage. 3. Transplanted MSCs could not differentiate to neuron and astrocyte without other interventions during 14 days after transplantation.

Animals ; Blood-Brain Barrier ; physiology ; Brain ; Cell Differentiation ; Cell Movement ; Disease Models, Animal ; Hypoxia-Ischemia, Brain ; therapy ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; physiology ; Rats ; Rats, Wistar

OBJECTIVETo study the acute toxicity of the water extracts (ERWE) and 60% ethanol extracts (EREE) from different processed products of Radix Polygalae (crude Radix Polygalae, licorice, and honey processed Radix Polygalae), thus providing scientific evidence for toxicity study of Radix Polygalae and its safe clinical application.

METHODSThe ERWE and EREE were prepared from different processed products of Radix Polygalae. Their contents of saponins were respectively determined. The poisoning condition and death of the mice administered with ERWE and EREE by gastrogavage were observed within fourteen days. The modified Karber's method was used to calculate LD50 and 95% confidence interval (CI).

RESULTSThe EREE of licorice processed Radix Polygalae had the maximum toxicity with highest content of saponins, while the ERWE of honey processed Radix Polygalae had the minimum toxicity with lowest content of saponins.

CONCLUSIONSDifferent processing methods have effects on the contents of saponins in Radix Polygalae. The experiment showed that the toxicity of Radix Polygalae is in direct proportion to the content of saponins. The higher the saponins contents, the higher the toxicity.

Animals ; Female ; Lethal Dose 50 ; Male ; Mice ; Mice, Inbred Strains ; Plant Extracts ; toxicity ; Polygala ; Saponins ; toxicity ; Toxicity Tests, Acute

This study was conducted to investigate the paclitaxel loaded by hydrazone bonds in poly(ethylene glycol)-poly(caprolactone) micelles (mPEG-PCL-PTX) on proliferation and apoptosis of human lung cancer A549 cells and its possible mechanisms of anti-tumor activity. The cell proliferation was measured with MTT assay. Flow cytometry were used to analyze the cell cycle. The cell apoptosis was analyzed using Hoechst/P staining. The expression levels of apoptotic genes expression in the mitochondrial apoptosis pathway were detected by RT-PCR and Western blotting, respectively. The mPEG-PCL-PTX could inhibit the proliferation of A549 cells and promote the apoptosis. The Bax, caspase-3 protein expression were increased while Bcl-2 protein expression was decreased in A549 cells. Results showed that the polymer containing hydrazone bond is non-toxic in vitro, the mPEG-PCL-PTX micelles can inhibit the proliferation and induce the apoptosis of A549 cells. Key words: paclitaxel; micelle; A549 cell; proliferation; cell cycle; apoptosis
Apoptosis ; Caspase 3 ; metabolism ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Micelles ; Paclitaxel ; pharmacology ; Polyesters ; Polyethylene Glycols ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-2-Associated X Protein ; metabolism