Objective To analyze the risk factors and prevention of non-diabetic mother neonatal hypoglycemia,in order to provide evidences for clinical diagnosis and avoid hypoglycemic brain injuries.Methods This was retrospective study using the method of case control.One hundred and eleven cases of non-diabetic mother neonatal hypoglycemia were enrolled in patients group,and 111 controls of normal blood glucose neonate at the same period were enrolled in control group.The risk factors of neonatal hypoglycemia and the results of neural development after hypoglycemia were analyzed.Results The risk factors of neonatal hypoglycemia were low-birth-weight infant(X2=4.066,P=0.044),small for gestation age infant (X2=21.125,P=0.000),congenital heart disease(X2=5.869,P=0.015),day-age≤3 days(X2=6.876,P=0.032),mother with pregnancy-induced hypertension(X2=6.618,P=0.010)or age<25 years old (X2=6.311,P=0.044).Full-term infants might be easier to show up hypogtycemic brain injuries than pre-term infants;the occurrence of hypoglycemic brain injuries correlated with the level of blood glucose and the lasting time of hypoglycemia.Conclusion Monitor blood glucose closely to the infant has risk factors of hypoglycemia,and discover and cure it on time,in order to avoid hypoglycemic brain injuries.

To obtain recombinant human β2-microglobulin (rhβ2M) with properties of good solubility and high purity from E. coli, prokaryotic expression conditions were optimized and protein purification was performed in this study. After testing the effect of different IPTG concentrations, temperatures and induction times on the production of rhβ2M, the optimum expression conditions were determined, i. e. joining IPTG to final concentration being 0.8 mmol/L and inducing time 6 h and at temperature of 25 degrees C. Under the optimum induction conditions, the ratio of soluble rhβ2M to soluble bacterial protein was 63.7%. After purified by Ni Sepharose 6 Fast Flow, the purity of rhβ2M achieved a greater value of 95%. Western blot analysis revealed that rhβ2M possessed the antigen property that specifically interacted with anti-β2M antibody.
Blotting, Western ; Escherichia coli ; metabolism ; Humans ; Recombinant Proteins ; biosynthesis ; Solubility ; beta 2-Microglobulin ; biosynthesis

Objective To investigate effects of ischemic postconditioning on the nitric oxide ( NO) and nitric oxide synthase ( NOS) in diabetic rat brain tissues .Methods Thirty Wistar rats were diabetic models induced by intraperitoneal injuction of stepto-zotocin (STZ), and randomly divided into three groups: Control group (normal, diabetic), cerebral ischemia group, and ischemic postconditioning ( I-POST) group.The rats of cerebral ischemia group and ischemic postconditioning group were made model of cere -bral ischemia by ligation carotid artery .Hematoxylin-eosin ( HE) was used to observe their pathological changes in control and diabetic groups.Enzyme-linked immunosorbent assay ( ELISA) method was used to detect the expression and changes of NO and NOS in the sera in each group .Western Blot method was used to investigate the expression and changes of NOS in the retinal tissues in each group .Results For I-POST group , brain tissue defects were decreased , neuronal cells were increased , serum inducible NOS ( iNOS) content was significantly lower than endothelial NOS (eNOS) and neuronal NOS (nNOS) ( P <0.05), brain tissue iNOS expression was significantly weaker than ischemia group ( P <0.05 ) and was not different from normal group ( P >0.05 ) .Conclusions Is-chemic postconditioning can protect the brain tissue of diabetic rats by inhibiting NOS activity especially iNOS .

Aim To investigate the effect of resveratrol on proliferation and differentiation in K562 cells. Methods K562 cells were treated with different con-centrations of resveratrol for 6d. The colony number of K562 cells was detected with semi-solid culture assay. Expression of GATA-1 and PU. 1 in K562 cells was re-spectively measured with immunocytochemistry and Western blot. Expression of differentiation related anti-gen, CD11b, CD14 and CD42b, was measured with flowcytometry on K562 cells. Results Resveratrol
could significantly decrease the colony number of K562 cells in a dose-dependent manner, and enhance the ex-pression of GATA-1,PU. 1,CD11b, CD14 and CD42b in K562 cells. Conclusion Resveratrol could inhibit the proliferation and induce differentiation of K562 cells via up-regulating the expression of GATA-1 and PU. 1 protein.

AIM:To investigate the effect of decitabine (Dacogen, DAC) on the proliferation and differentia-tion of K562 cells.METHODS:The K562 cells were treated with different concentrations of DAC .The colony formation ability of the cells was detected by the colony formation assay with semi-solid culture .The cell viability was detected with MTT assay.The morphologic features were observed under inverted microscope with Wright ’s staining.The changes of the cell cycle distribution and the expression of CD 11b and CD42b were analyzed with flow cytometry .The protein expression of CDK2, cyclin E1, P27, GATA-1 and PU.1 in the K562 cells was determined by Western blot .RESULTS:DAC signifi-cantly decreased the colony number of the cells and cell viability in a dose-dependent manner .The morphological changes of the cells displayed partial differentiation .After treated the K562 cells with DAC for 72 h, the cell proportion in S phase was obviously decreased , while the cell proportion in G 2/M phase was obviously increased in a dose-dependent manner . After treated the K562 cells with DAC for 7 d, the percentage of CD11b and CD14 positive cells was further elevated , and the protein expression of P27, GATA-1 and PU.1 was increased.However, the protein expression of CDK2 and cyclin E1 was decreased .CONCLUSION:DAC inhibits the proliferation and induces differentiation of the K 562 cells via regulation of cell cycle .

Aim To investigate the effect of bufalin on proliferation and expression of WT1 in K562 cells. Methods The colony number of K562 cell was detec-ted with semi-solid culture assay. The cell cycle was measured by flowcytometry, and the expression of WT1 was observed with immunocytochemistry. Subcutaneous tumor models established by K562 cells in BALB/C nu/nu mice were divided into three groups, including model group, bufalin group and positive control group. After 21 days, the subcutaneous tumors were removed for calculating the inhibitory rate of tumor growth. HE staining and immunohistochemistry were used to ob-serve the morphological changes and the expression of WT1 . Results ① Bufalin could significantly decrease the colony number of K562 cell, arrest it at G0/G1 phase and down-regulate its expression of WT1 in a dose-dependent manner. ② Compared with the model group, the tumor inhibitory rate was much higher, while the volume and the weight were obviously lower in the other two groups. ③Bufalin could induce apop-tosis, necrosis, hemorrhage and fibrosis with HE stai-ning, and down-regulate the expression of WT1. Con-clusion Bufalin could inhibit the proliferation, arrest the cell cycle at G0/G1 phase and down-regulate the expression of WT1 in vitro. Bufalin could inhibit the tumor inhibitory rate, the volume and the weight of the subcutaneous tumors, induce apoptosis, necrosis, hemorrhage and fibrosis with HE staining and down-regulate the expression of WT1 .

Abstract: Cell death is a fundamental biological phenomenon that is essential for the survival and development of organisms. Cell death can be either a spontaneous programmed process by the host or an accidentally triggered process. According to the different signaling pathway activated by various stimulates, programmed cell death exhibits the lytic or non-lytic morphology. For example, apoptosis, a typical non-lytic form of cell death, exhibits cell shrinkage and induces the formation of apoptotic bodies. Pyroptosis mediated by cysteine-containing aspartate-specific protease-1/11 (caspase-1/11) and necroptosis can induce inflammatory reactions and promote cell lysis to release inflammatory cytokines via triggering the pore-forming mechanism of the cell membrane, representing a typical modes of lytic cell death. In addition, the release of reactive oxygen species caused by the damaged mitochondria may further trigger ferroptosis during the pathogen infection. Programmed cell death can play an immune defensive role by eliminating infected cells and intracellular pathogens and stimulating the innate immune response through the resulting cell corpses. Here, we discuss the molecular mechanisms of five programmed cell death pathways: apoptosis, pyroptosis, ferroptosis, necroptosis and PANoptosis. We describe their roles in the innate immune defense against bacterial infections and give a brief statement of the interactions between the different programmed cell death, hoping to provide new insights for in-depth study of the pathogenic mechanisms of infectious diseases.

Objective To investigate the influence of ionic strength on the stability of the methotrexate-loaded dendrimer nanoparticles. Methods The influences of different ions (Na+,Cl-) and different concentrations of sodium chloride on the stability of the nanoparticles were studied. The particle size was measured by dynamic light scattering(DLS) and drug-loading content was determined by high-performance liquid chromatography (HPLC) in order to evaluate the stability. Results The Cl- was finally verified to play an important role in stabilizing the nanoparticles and the effective concentration of the sodium chloride was recommended to be below 1. 80% . Conclusion The recommended concentration (less than 1. 80% ) of the sodium chloride significantly improves the stability of the nanoparticles and benefits for long term storage.

AIM: To study the protective effects of Naokangning (Rhizoma Chuanxiong, Radix salviae miltiorrhizae, Hirudo, etc.) against cerebral ischemia. METHODS: We observed the effect of Naokangning on mice's resistance to cerebral ischemia when bilateral common carotid arteries and vagus nerves were ligated and hypoxia under normal pressure and airtight circumstance; With the model of partial cerebral ischemia by blocking rats'middle cer ebral artery (MACO):the effects of Naokangning on the activity of superoxide dismutase (SOD) and creatine kinas e(CK), the content of malondialdehyde (MDA) and nitric oxide (NO) were measured. RESULTS: Naokangning significantly raised mice's ability of anti-cerebral ischemia and prolonged span of life in hypoxia, Moreover, it also obviously improved the activity of SOD, reduced content of MDA in cerebrum, content of NO and activity of CK in blood serum after ischemia. CONCLUSION: Naokangning could strikingly protect brain caused by cerebral ischemia.

Objective To evaluate the clinical outcome of NB09 (China Pediatric Neuroblastoma cooperative group 09) protocol on children with high-risk and ultra-high risk neuroblastoma. Methods The clinical and follow-up data of pa?tients who suffered from high-risk (n=7) and ultra-high risk (n=31) neuroblastomas and admitted in Tumor hospital of Tian?jin Medical University between January 2009 to January 2013 were retrospectively reviewed (27 boys and 11 girls). The age at diagnosis was 19-160 months (median age was 36.5 months). In the high risk group, patients were evaluated and operated after 4 to 6 circles of neoadjuvant chemotherapy. In ultra-high risk group, patient received chemotherapy before and after op?eration, then autologous stem cell transplantation and tumor bed radiotherapy. After chemotherapy, retinoic acid treatment was given to patients in ultra high risk group as in high risk group. Results At the end of treatment, 25 patients achieved complete remission; 5 patients achieved partial remission; 3 patients were in stable disease;5 patients were deteriorating in their conditions which lead to 2 deaths. In total, the response rate reaches upto 86.8%. By the end of follow up, 15 patients had a disease-free-survival, 9 patients survived with tumor, 7 died from recurrence and 7 died from deteriorating conditions. Survival time ranged from 6 to 52 months (median survival 25.5 months). The 1-, 2- and 3-year overall survival were 91.7%, 64.5%and 57.3%respectively. Kaplan-Meier curve and Log-rank test showed no statistical significance between high risk and ultra-high risk neuroblastomas. Conclusion The outcome of NB09 protocol for high risk and ultra-high risk neuroblastoma was preliminary affirmed. It is worthy of further clinical verification.