The human interleukin-24 gene (hIL-24),alse referred to as melanoma differentiation associated gene-7 (MDA-7), is a novel cancer-selective apoptosis-inducing eytokine gene with broadspectrum antitumor activity. Multiple data have demonstrated that treatment of tumors with a combination of IL-24 and other strategies such as drug therapy, radiation, dual gene therapy and so on, caused synergistic antitumor effects. These results will provide a impetus for further combination therapy in various tumors.

Objective To investigate the synergistic proliferation-inhibiting and apoptosis-inducing effects of recombinant human-derived interleukin-24 (rhIL-24) and recombinant human-derived decorin (rhDCN) on human hepatocellular carcinoma cells HepG2.Methods Cellular growth and morphological changes of HepG2 cells were observed under the inverted microscope at 48 h after being transiently transfected with pcDNA3.1 (+)-IL-24 and pcDNA3.1 (+)-DCN by Lipofectamine.The proliferation-inhibiting effects of IL-24 and DCN on HepG2 cells,respectively and jointly,were observed with MTT assay at 24 h,48 h and 72 h post-transfection.Apoptosis and cell-cycle of HepG2 cells were analyzed by flow cytometry at 48 h post-transfection.Results Compared to control groups,the cells of target gene groups presented typically changes of proliferation inhibition and apoptotic morphology,which occurred obviously in co-transfection group.The results of MTT assay showed that at 48 h and 72 h post-transfection,the profiferation-inhibiting rates in the group of cells co-transfected with IL-24 and DCN were (31.88±6.57) ％ and (36.83±3.76) ％,respectively,displaying significant difference with those of other groups (P ＜ 0.01).The results of flow cytometry showed that IL-24 and DCN can induce HepG2 cells apoptosis to some extent.Compared to the early apoptosis rate of cells of control groups,plasmid (2.98±0.72) ％,blank cell (3.50±0.92) ％,IL-24 (20.01±1.08) ％ and DCN (22.20±0.91) ％,a statistically remarkable apoptosis rate,(32.56±0.90) ％,can be seen in the group of cells treated with IL-24 and DCN jointly (P ＜ 0.01).The result of cell cycle analysis revealed that,compared to control groups,the proportion of cells was higher in the phase of G2/M in the IL-24 group (11.24±0.35) ％ and in the phase of G0/G1 in the DCN group (77.93±0.67) ％.The proportions of cells in the phases of G2/M increased to (71.36±0.60) ％ and that of G0/G1 statistically increased to (10.39±0.67) ％ in the group of cells co-transfected with IL-24 and DCN (P ＜ 0.01).Conclusions Combinatorial treatment of HepG2 cells with IL-24 and DCN can exert stronger synergistic proliferation-inhibiting effect and apoptosisinducing activity-in comparison to single therapies.IL-24 and DCN can induce cell cycle arrest on HepG2 cells,occurred in the phase of G2/M and G0/G1,respectively.Promoting effect of cell cycle arrest in the phase of both G2/M and G0/G1 can be seen on HepG2 cells co-transfected with IL-24 and DCN,which maybe the possible mechanism of the synergistic proliferation-inhibiting and apoptosis-inducing effect.

Objective To investigate the effects and mechanisms of propofol and dexmedetomidine on neurites and synapses of hippocampal neurons neonatal rats, in vitro. Methods Hippocampal neurons of neonatal Sprague-Dawley rats were cultured 6 days, in vitro and were divided into control group (Group C), solvent of propofol group (Group S), propofol group (Group P), dexmedetomidine group (Group D),propofol and dexmedetomidine group (Group PD), and yohimbine group (Group Y). All groups were cultured for 24 h further. Neuron morphology and the expression of protein were measured. Results After exposing to propofol, we found that the mean total length of neurites of primary cultured hippocampal neurons and synapses and the expression of BDNF, TrkB and CRMP-2 protein were reduced; dexmedetomidine played a protective role. Moreover, yohimbine, partly inhibited neuroprotection of dexmedetomidine. Conclusions Propofol decreases the development of neurites and synapses of hippocampal neurons neonatal rats, in vitro, and dexmedetomidine provides a protective effect on propofol by up-regulating the expression of BDNF, TrkB and CRMP-2. The effect, partly has a concern aboutα2-adrenergic agonist.

Objective To evaluate the effect of isoflurane on the apoptosis of SH-SYSY cells transfected with APPsw gene and the role of inositol 1,4,5-triphosphate (IP3) recepters.Methods The SH-SYSY ceils transfected with APPsw gene were seeded in culture flasks with the density of 1.2 × 104/cm2.The cells were randomly divided into 4 groups (n =6 each):control group (group C),IP3 receptor antagonist group (group Ⅹ),isoflurane group (group Ⅰ) and isoflurane + IP3 receptor antagonist group (group Ⅰ + Ⅹ).After the cells were cultured for 24 h and attached to the wall,the cells were cultured routinely in group C,and Xestospongin C 100 nmol/L (IP3 receptor antagonist) was added to DMEM culture medium in groups X and Ⅰ + X,and 30 min later the cells were exposed to 1.2 ％ sevoflurane for 8 h in groups Ⅰ and Ⅰ + X.The cells were collected for examination of the ultrastructure and for determination of cell apoptosis,intracellular free calcium ion concentration [Ca2 +] i (by flow cytometry) and expression of IP3 receptor protein (by Western blot).The apoptosis rate was calculated.Results Compared with group C,there was no significant change in the apoptosis rate,[Ca2 +]i or IP3 receptor protein expression in group Ⅹ (P ＞ 0.05),while the cell apoptosis rate and [Ca2 +] i were significantly increased and IP3 receptor protein expression was up-regulated in groups I and Ⅰ + Ⅹ (P ＜ 0.05 or 0.01).Compared with group Ⅰ,cell apoptosis rate and [Ca2+]i were significantly decreased and IP3 receptor protein expression was down-regulated in group Ⅰ + Ⅹ (P ＜ 0.01).The pathological changes of the cells happened in groups Ⅰ and Ⅰ + Ⅹ,and the pathological changes were severer in group Ⅰ than in group Ⅰ + Ⅹ.Conclusion Isoflurane can induce apoptosis of SH-SY5Y cells transfected with APPsw gene through increasing [Ca2+]i and up-regulating IP3 receptor protein expression.

Objective To investigate the suppression effect, the apoptosis and TGF-β_1 mRNA expression of rhDCN and dororubicin(ADM) on leukemic K562 cell line. Methods K562 cells in Logarithmic growth phase were divided into Saline group, pcDNA3.1 (+)-DCN group, ADM group, and pcDNA3.1 (+)-DCN-ADM group. Morphology change of cell was detected by Wright stain, cell proliferation activity was assessed by MTT. The apoptosis index of K562 cells was assessed by FCM, and TGF-β_1 mRNA of cell was assessed by RT-PCR. Results Wright stain showed that more pronounced morphological apoptosis changes of K562 cells in combined group. MTT method results showed that the proliferation inhibition rate of the combined group was (61±1.32) % higher than that of individual intervention group [DCN group, (20±1.90) %; ADM group, (47±1.04) %](P <0.05). FCM results showed that the apoptosis index of the combined group was (61.30± 0.9) %, higher than that of Individual intervention group [DCN group, (28.25±1.3) %; ADM group, (31.85± 1.5) %](P <0.05). TGF-β_1 mRNA synthesis of combined group was significantly decreased. Conclusion rhDCN can markedly enhance cytotoxicity of ADM on K562 cells, and the mechanisms of apoptosis may be due to down-regulation of TGF-β_1 mRNA. Specific mechanisms will be further studied.

Balantidium coli human infection predominantly occurs in tropical and subtropical regions in the world. Human case is extremely rare in China. This report details a case of B. coli infection in a 68-year-old man in China, who presented with history of abdominal pain, tenesmus, diarrhea with blood and was diagnosed as B. coli-caused dysentery. Our case indicates possible occurrence of Balantidium coli-related disease in cooler climates. This case is presented not only because of its rarity but also for future references.

Objective:To evaluate the effect of transcutaneous auricular vagus nerve stimulation (taVNS) on tourniquet-induced hypertension (TIH) in the patients undergoing anterior cruciate ligament reconstruction.Methods:Seventy-four patients of either sex, aged 18-60 yr, of American Society of Anesthesiologists Physical Status classification I or II, with body mass index of 18-30 kg/m 2, undergoing elective anterior cruciate ligament reconstruction under general anesthesia combined with preoperative femoral nerve block, were divided into 2 groups ( n=37 each) using a random number table method: sham stimulation group (group SS) and group taVNS. Group SS received stimulation on the ear lobe and the tail of the helix of the left ear. Group taVNS received stimulation on the cymba concha and the earlobe of the left ear. Both groups received stimulation from 1 h before induction of anesthesia until the end of the procedure (frequency of 30 Hz, pulse width of 300 μs, and amplitude of the strongest current that could be tolerated by the patient in the absence of pain). The tourniquet inflation pressure was 280 mmHg, with an inflation time of 60-90 min. Systolic blood pressure, diastolic blood pressure and heart rate were recorded before tourniquet inflation to assess the development of intraoperative TIH. The consumption of intraoperative propofol, remifentanil, nitroglycerin, esmolol, norepinephrine and atropine was recorded, and the occurrence of postoperative nausea and vomiting, skin itching and headache and dizziness was also recorded. Results:Compared with group SS, the incidence of TIH and the number of patients used nitroglycerin were significantly reduced ( P<0.05), and no significant changes were found in the other parameters in group taVNS ( P>0.05). Conclusions:taVNS can decrease the occurrence of TIH in the patients undergoing anterior cruciate ligament reconstruction.

Objective To assess the value of flow cytometry in the diagnosis of postoperative infection following renal transplantation. Methods According to postoperative imaging findings and laboratory examination outcomes, 51 recipients undergoing the first renal transplantation were divided into the bacteria (n=33), fungus (n=9) and BK virus (n=9) groups. Twenty-eight recipients with stable conditions after renal transplantation were assigned into the stable group. Flow cytometry was adopted to detect the percentage and absolute counting of lymphocyte subpopulation in the peripheral blood of recipients in each group. Renal function, percentage and absolute counting of lymphocyte subpopulation in the peripheral blood were statistically compared among different groups. Receiver operating characteristic (ROC) curve was drawn to evaluate the diagnostic value of the percentage and absolute counting of lymphocyte subpopulation in infectious diseases after renal transplantation. Results Compared with the stable group, the serum creatinine (Scr) and blood urea nitrogen (BUN) levels in the bacteria, fungus and BK virus groups were significantly up-regulated, respectively (P=0.035, 0.007, 0.024; 0.037, 0.006, 0.032). Compared with the stable group, the percentage of CD16+CD56+natural killer (NK) cells was significantly declined in the bacterial (P=0.036) and fungus groups (P=0.015), and the proportion of CD4+/CD8+T cells was dramatically decreased in the fungus group (P=0.004). Compared with the bacterial group, the percentage of CD3+CD8+T cells was significantly elevated (P=0.013 and 0.008), the proportion of CD3+CD4+T cells was considerably declined (P=0.003 and 0.010), and the percentage of CD4+/CD8+T cells was significantly declined (P=0.003 and 0.005) in the fungus and BK virus groups. Compared with the stable group, the quantity of CD3+T cells, CD3+CD8+T cells and CD16+CD56+NK cells was significantly declined in the bacterial, fungus and BK virus groups, respectively (P=0.025, 0.002, 0.003; 0.015, 0.005, 0.006; 0.001, 0.001, 0.031). In addition, the quantity of CD3+CD4+T cells was considerably decreased in the fungus and BK virus groups (P=0.001, 0.003). The quantity of CD19+B cells was significantly reduced in the BK virus group (P=0.019). Compared with the bacterial group, the quantity of CD3+CD4+T cells was considerably lower in the fungus group (P=0.023). ROC curve analysis revealed that the quantity of CD3+CD4+T cells [area under curve(AUC)=0.8492] and CD16+CD56+NK cells (AUC=0.8889) yielded relatively high accuracy in the diagnosis of fungal infection. The quantity of CD3+T cells (AUC=0.8472), CD3+CD4+T cells (AUC=0.8452) and CD19+B cells (AUC=0.8115) yielded relatively high accuracy in the diagnosis of BK virus infection. Conclusions Flow cytometry detection of the lymphocyte subpopulation in peripheral blood can evaluate the immune function of patients. Absolute counting of lymphocyte subpopulation can directly assess the degree of immunity. These two combined parameters provide guiding significance for the diagnosis and differential diagnosis of infectious diseases in recipients after renal transplantation.

Pseudorabies virus(PRV)is the etiological agent of pseudorabies in pigs,which is char-acterized by dyspnea,reproductive disorders,and neurological diseases,and it spreads widely a-round the world.Since 2011,the newly emerged PRV variants have resulted in poor immunity pro-tection of traditional vaccine strains,and the original method of vaccine strain preparation is time-consuming and labor-intensive.Therefore,it is urgently needed to develop an efficient screening method of the vaccine strain at present.Using CRISPR/Cas9 gene editing technology in this study,two single guide RNAs(sgRNA)were designed targeting the virulence gene TK of PRV variant strain CH/JX/2016,and then the enhanced green fluorescent protein the reporter(EGFP)gene was inserted at the TK locus by a homologous repair plasmid.After multiple rounds of plaque puri-fication,the rPRV-ΔTK/EGFP strain was obtained.The results showed the cleavage efficiency of the two sgRNAs was extremely high.The preparation of rPRV-ΔTK/EGFP strain was succeed af-ter only three rounds of purification,and the EGFP expressed normally.The CRISPR/Cas9 system can edit the PRV gene simply,rapidly,and efficiently,and exhibits great potential in the construction of vaccine candidate strains.Meanwhile,the rescued rPRV-ΔTK/EGFP strain not only could be used as a tracer strain in PRV variant infection progresses,but also for subsequent antivi-ral drug screening.