Objective To explore effects of human embryonic stem cells ( hESCs) on proliferation, invasion and migration of SK-Hep1 human hepatoma cells in the co-culture of micro environmen of hESCs and SK-Hep1 . Methods Single cultured SK-Hep1 cells were served as control group while SK-Hep1 which non-contact co-cul-tured with hESCs was regarded as experimental group .The proliferation ability of SK-Hep1 was measured by MTT method; invasion and migration ability of SK-Hep1 cells were detected by Transwell chamber method;the nucle-us variation and cell apoptosis of SK-Hep1 were detected by Hoechst33258 chromosome and flow cytometry. Results The proliferation of SK-Hep1 cells in the experimental group was obviously inhibited as compared with control group ( P<0.05 );the number of SK-Hep1 cells which passed through the Transwell chambers were sig-nificantly reduced as compared with control group in invasion and migration experiment ( P <0.05 ); more nucleus pycnosis and deformation appeared in experimental group than that in control group .And apoptosis rate of SK-Hep1 cells in the experimental group was significantly higher than that of in the control group ( P<0.05 ) .Conclusions Human embryonic stem cells have inhibitory effect on human hepatoma cell line SK-Hep1 .

Animals ; Male ; Methyl Chloride ; therapeutic use ; Myocardium ; pathology ; Organothiophosphorus Compounds ; poisoning ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the protective effect of Shenfu Injection against ischemia-reperfusion (I/R) injury due to pancreas transplantation in rats, and explore its possible mechanism.

METHODSSix normal SD rats with sham operation were taken as the normal control group, 24 steptozozin-induced diabetic SD rats were randomly divided into 4 groups, with 6 in each group. Except I/R group, the rats in the other groups were intravenous injected with Shenfu Injection (SF,10 mg/kg), Hongshen Injection (HS, 9 mg/kg) and Fuzi Injection (FZ 1 mg/kg) respectively at the day and 30 minutes before pancreas transplantation performed in the SF group, HS group and FZ group, respectively. At the same time, rats in the normal control group and in the I/R group were intravenously injected the same volume of normal saline. The blood glucose was detected before and after reperfusion, and 2 hours later after reperfusion, the contents of serum nitric oxide (NO) and tumor necrosis factor-alpha (TNF-alpha), the concentrations of malondialdehyde (MDA) , superoxide dismutase (SOD) , and myeloperoxidase (MPO) in the transplanted pancreas tissues were detected. The cell apoptosis of the transplanted pancreas tissue was determined by TUNEL, and the bcl-2 and Bax protein expression was determined by Western blot.

RESULTSAfter reperfusion, the levels of blood glucose and TNF-alpha decreased and the concentration of NO increased in the SF group, HS group and FZ group, compared with those in the I/R group. The activity of SOD, bcl-2 expression and the ratio of bcl-2 and Bax were higher, while the content of MDA, the activity of MPO, apoptotic indexes, and Bax expression were lower in the SF group, HS group and FZ group than those in the I/R group.

CONCLUSIONShenfu Injection can protect L/R injury due to pancreas transplantation in rats, the possible mechanism may be related to promoting activity of SOD, increasing synthesis of endogenous NO, decreasing the excretion of TNF-alpha, alleviating conglutination and aggregation of polymorphonuclear neutrophils (PMNs) in pancreas, as well as up-regulating Bcl-2 gene expression and down-regulating the Bax gene expression.

Animals ; Cell Aggregation ; drug effects ; Diabetes Mellitus, Experimental ; enzymology ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Injections ; Malondialdehyde ; metabolism ; Nitric Oxide ; blood ; Pancreas ; drug effects ; metabolism ; Pancreas Transplantation ; adverse effects ; Protective Agents ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; etiology ; prevention & control ; Superoxide Dismutase ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

AIM:To establish a method for obtaining specific cells in solid tumor tissue by sorting of CD11b + myeloid cells in hepatic metastases from colorectal cancer.METHODS:Tumor tissues were prepared into single cell suspension by mechanical method combined with enzyme digestion,and then the CD11 b + myeloid cells were isolated by flow cytometry.The sorted cells were identified by immunocytochemistry.The viability and morphologiy of the sorted cells were evaluated by Giemsa and Typan blue staining.The cell purity was evaluated by flow cytometry.RESULTS:Sufficient numbers of CD11b+cells with high purity were isolated by sorting with flow cytometry from the single cell suspension prepared by mechanical and enzyme digestion.The purity of the cells was confirmed by statistical analysis (P ＜ 0.05).The positive rates of the cells before and after sorting were significantly different (P ＜0.01).The positive cells were verified by immunocytochemical method.Meanwhile,the sorted cells had complete morphology and good activity.CONCLUSION:The CD11b + myeloid cells in solid tumor tissue can be isolated by flow cytometry from the machine-enzyme digestion suspension with high purity,good activity and complete morphology.

Objective To provide theoretical and experimental proof for selecting and implying Tourette's syndrome(TS) animal models, validities of four TS models induced by chemical factors were compared. Methods Four TS models,namely AMP model,APO model,DO1 model and IDPN model were built up by using different chemical modeling agents. Through detecting spontaneous movement, climbing time and monoamine transmitters levels in striatum, four TS animal models were compared and evaluated from three levels of validities-face, prediction,construct. Results Compared with control group, spontaneous movement times raised ( t = 4. 746, P =0. 000) and level of DOPAC ( (0.99 ± 0. 177 ) ng/mg) in striatum increased (P = 0.029 ), and level of NE in striatum decreased in AMP model group( (0.11 ± 0.033 )ng/mg, P = 0.012). Compared with control group, climbing time prolonged (P = 0. 004) and levels of DA ( ( 10. 19 ± 1.23 ) ng/mg), 5-HT ( ( 0. 54 ± 0.08 ) ng/mg) in striatum raised(P=0. 019, P=0. 002),at the same time ,levels of DOPAC( (0.63 ±0.11 )ng/mg),HVA ((0.45 ±0.04 ) ng/mg) in striatum reduced (P ＜ 0.01 ) in APO model group; Compared with control group, levels of DA ( ( 13.66 ± 1.55 ) ng/mg), DOPAC( (0.80 ±0. 11 ) ng/mg), HVA( ( 1.04 ± 0.14) ng/mg) grew downwards in striatum of DOI model mice(P=0.029,P=0.001, P= 0.004). Compared with control group, level of 5-HT in striatum increased in IDPN300 group ( (0.77 ± 0.09) ng/mg, P = 0.031 ). ConclusionFace validity of AMP model is temporal and that of IDPN model is steady and persistent. AMP model,APO model and DOI model possess predictive validity. AMP model,APO model,DOI model and IDPN model have potentiality of becoming construct validity model.

As the increasing population aging,the incidence of age-related macular degeneration is increasing,blinding rate also increasing,so it is very important for the early diagnosis and treatment of age-related macular degeneration.There are many methods to check the aging macular degeneration,such as fundus angiography,optical coherence tomography (OCT),visual field and multifocal electroretinogram (mfERG).In recent years,many emerging ophthalmic methods have emerged and are gradually applied to clinical diagnosis,including optic coherence tomography angiography (OCTA).The function of these methods has its unique advantages,but there are also limitations.This paper will review these existing methods.

Objective To evaluate the effects and underlying mechanisms of crocin on glutamate-induced apoptosis of retinal ganglion cells (RGCs) by affecting extracellular calcium influx.Methods Primary rat retinal ganglion cells were isolated and stimulated with glutamate at concentrations of 0.1 mmol/L and 1 mmol/L for 24 h or 48 h,respectively,to establish apoptosis model of RGCs.Afterwards,crocin of different doses (0.1,1.0 and 3.0 μmol/L) was used to treat the glutamate-induced RGCs for 12 h;then cell apoptosis was detected by Annexin V-FITC/PI staining.The intracellular calcium concentration was determined by FIuo-3/AM fluorescent labeling.Western blot was used to examine the effect of crocin on Ca2+-mediated apoptotic signal molecules calpain and CaMKII.The mitochondrial membrane potential was detected by JC-1 staining and mitochondrial apoptosis-related signaling molecules Caspase-3,Caspase-9 and Bcl-2/Bax were evaluated by Western blot,respectively.Results In comparison with the untreated controls,the cell apoptosis of RGCs exposed to 0.1 mmol/L of glutamate for 24 h did not significantly change (P＞ 0.05).However,apoptosis rate of the cells reached (43.050 ± 2.616) ％ when the stimulation time lasted for 48 h and showed a significant increase (P＜0.01).Treatment with higher-dose glutamate (1 mmol/L) significantly increased apoptosis of RGCs at 24 h (46.450±1.061)％ and 48 h (45.500±3.253)％ compared with the controls (P＜0.01).RGCs were induced by 1 mmol/L of glutamate for 12 h,followed by the treatment with crocin at concentrations of 0.1,1.0 and 3.0 μmol/L,respectively.Each dose of crocin could significantly inhibit cell apoptosis in the dose-dependent manner (P＜0.01).In addition,crocin at 1.0 μmol/L blocked glutamate-induced extracellular calcium influx,inhibited the expression of calcium-dependent proteins Calpainl and CaMK Ⅱ.Moreover,crocin at the dose of 1.0 μmol/L also increased mitochondrial membrane potential,suppressed the expressions of Caspase-3 and Caspase-9,and elevated Bcl-2/Bax ratio.Cornclusion Crocin inhibits glutamate-induced apoptosis of retinal ganglion cells through suppressing extracellular calcium influx,thereby blocking calcium-dependent and mitochondria-dependent apoptosis signaling pathways.

Objective:To study the inhibitory effect of human embryonic stem cells on the HepG2 cells in vitro.Methods:The co-culture system of Human embryonic stem cells (H9) and liver cancer HepG2 cells was established.The effect of H9 on the biological behavior of HepG2 cells was observed by microscope,the flow cytometry was used to detcct the apoptosis of tumor cells and the cell cycle alteration.Transwell assay was used to detect the migration and invasion of tumor cells.Gene microarray technique was used to examine the change of gene expression profile of HepG2 cells.Results:In the process of co-culture,the growth of hepatoma cells was inhibited.With the extension of the culture time,cells decreased gradually,and occurred signs of aging or apoptosis.Flow cytometry test results showed that the apoptosis rate of hepatoma ceils was significantly increased,and the cell cycle was blocked in the G0/G1 phase.Transwell test results showed that the invasion and migration of HepG2 cells were decreased.The gene chip results showed that the whole genome expression profile of HepG2 cells had a significant change.Conclusion:The human embryonic stem cells had an inhibitory effect on HepG2 cells in vitro.

OBJECTIVE:To provide reference for investigating the nutrition situation of the inpatients in departments of spinal surgery and minimally invasive spinal surgery and promoting the clinical rational use of nutritional support drugs. METHODS:Des-ignated continuous sampling was used to select the inpatients that fit the conditions in the departments of spinal surgery and mini-mally invasive spinal surgery in our hospital from Jan. to Dec. 2013,and the nutritional risk screening 2002 was used to investigate the patients’nutritional risk at admission and upon discharge,the nutritional support during hospitalization were recorded. RE-SULTS:In the 432 enrolled patients,the overall incidence of nutritional risk was 11.57% at admission,12.40% in spinal surgery and 10.44% in minimally invasive spinal surgery;and the overall incidence of nutritional risk was 19.44% upon discharge, 23.60% in spinal surgery,with statistical significance when compared with admission(P<0.05),while 13.74% in minimally inva-sive spinal surgery,with no statistical significance when compared with admission(P>0.05). The nutritional support rate of 50 pa-tients with nutritional risk at admission was 88.00%,14.14% of 382 non-risk patients still received a redundant nutritional support. The 44 patients with nutritional risk who received nutritional support had the average calories intake of(9.84±8.10)kJ/(kg·d),in-cluding 10.16% lipids and 9.55% protein;all patients who received nutritional support found no patients with enteral nutrition sup-port. CONCLUSIONS:Inpatients in departments of spinal surgery and minimally invasive spinal surgery suffer a lower incidence of nutritional risk at admission,and a higher incidence of nutritional risk in the former one upon discharge. While some patients who received nutritional support show no indication,and administrations of nutritional support are still debatable,the use of drug is irrational.

A 49-year-old male was admitted to Peking Union Medical College Hospital presented with fever for more than half a year. The patient was diagnosed as Sjogren′s syndrome at local hospital. After oral prednisone 60 mg per day was given, the fever alleviated, but recurred after prednisone tapered to 40 mg/d. Both blood culture and stool culture were positive for Salmonella enteritidis. Antibiotics including ceftazidime, ceftriaxone, cilastatin-imipenem were sequentially administrated for 4 weeks, yet not effective. Although there were not respiratory symptoms or certain abnormalities on high-resolution chest CT, arterial blood gas indicated hypoxemia. Serum lactate dehydrogenase and β2 micro-globulin were elevated, and the lung function test demonstrated significant impairment of diffusion function. Positron emission tomography-computed tomography (PET/CT)scan suggested that high fluorodeoxyglucose uptake was diffusely seen in both lungs. The patient was finally diagnosed as pulmonary intravascular large B-cell lymphoma (IVLBCL) by transbronchial lung biopsy. This case aims to emphasize the differentiation diagnoses of pulmonary intravascular lymphoma from common situations.