Abstract:Objective:Microbubble is a kind of traditional contrast agent for ultrasound imaging and also one of the most important probe carriers for ultrasound molecular imaging. The developing trend of molecular probes on microbubbles is to multimodal,multifunctional and intelligent. Now there have been lots of novel and multifunctional molecular probes developed based on microbubbles in recent years. This review summarized the design and application of different types of ultrasound molecular probes. It includes multimodal molecular probes of various imaging modes,multifunctional molecular probes of integration of diagnosis and therapy,stimulated response ultrasound molecular probes triggered by special pathological conditions and enhanced tissue penetration ultrasound molecular nano-probes.

Abstract:Objective:To develop a simple,low cost,portable and easy to realize automation pathogen nucleic acid detection system to meet the rapid on-site testing requirements. Methods:An enclosed cartridge was designed for integrated pathogen nucleic acid testing and creatively manufactured using 3D printing technology that speeded up the development cycle and saved the cost of the whole system. On this basis,a fully automatic portable system was developed,which now is able to automatically and integratedly realize single or multiple nucleic acid detection. Results:Various tests of this instrument were carried out,induding airtightness of cartridge and pipetting performance,blending performance,etc. All test results met the requirements of pathogen detection. At last,take pertussiss for example,the practical applications of nucleic acid detection showed that the cartridge system can complete nucleic acid testing automatically. Conclusion:This system can successfully and automatically complete the integration testing process of infectious diseases from sample input to result output. The experimental results are accurate and reliable.

Abstract:Wearable technology is one of the research hotspots,which can be used in real-time monitoring of hospital patients and family health caring,and it has broad application foreground and huge marketing potentiality. Wearable technology puts forward higher requirements on the wearability of devices and the reliability and security of data transmission. There are some problems needed to be studied and solved in the development of this technology. In this paper,the research status and industrialization progress of wearable technology are reviewed,and the studies and development of wearable technology in the future are prospected.

Abstract:Objective:The hospital space positioning system for patients and the medical staff,which is used to achieve the goals of spatial orientation,is a new application of ward management. Methods:The system uses the RFID tag as the core and the RF8506 as the reader unit,which constitutes the ZigBee terminal device with the CC2530 wireless single-chip computer. The terminal devices were placed in the ward entrance to recorded whether the patients went out of the room. The network was set up by coordinator,and positioning and display function were realized by computers. Conclusion:The space positioning system can be seen as a fuzzy space positioning. It is different from video surveillance and the existing flat pinpoint ways such as GPS,because it protects the patient’s privacy and it achieves the regional location of floor space. From above,the system can simplify the management of hospital ward and reduce the workload of medical staff.

Abstract:Objective:To generate human monoclonal antibodies(mAbs)against the rabies virus glycoprotein(RVG)by immunizing the mice carrying human immunoglobulin transloci, and further to identify their characters. Methods:The human IgM transgenic mice were immunized with the rabies virus glycoprotein. The human anti-RVG hybridoma cell lines were screened and produced by using the classical hybridoma technique. The specificity and isotype of mAbs were determined by the double antibody sandwich enzyme- linked immunosorbent assay(ELISA). Also,the binding properties with the inactivated rabies virus CVS-11 were analyzed. Results:Five hybridoma cell lines were established,they can stably produce human anti-rabies virus IgM monoclonal antibodies(5D1,6H11,9A3,15D6,and 19E6). These mAbs specifically recognized the r-RVG and three human anti-r-RVG mAbs specifically combined with the inactivated rabies virus CVS-11 strain. Conclusion:Hybridoma cell lines were successfully established,and can stably produce human anti-rabies virus IgM monoclonal antibodies. Human anti-r-RVG mAbs could specifically combine with the inactivated rabies virus CVS-11 strain. This may be a foundation for further development of vaccine to prevent and control rabies.

Abstract:Objective:To explore the potential protective effects and mechanism of isocitric acid in post-thawing sperm. Methods:After routine semen analysis,semen samples from 16 fertile males were selected. Every sample was equally divided into five groups. One group was mixed with routine protectant as negative control,and the other groups were mixed with modified protectant containing different concentration of isocitric acid. After and before thawing,the sperm samples were subjected to CASA for analysis and flow cytometry for mitochondrial transmembrane potential,respectively. Results:The percentage of forward movement sperm in G2 and G3 group was higher than that of the control group(P < 0.05 and P < 0.01). Sperm concentration of G3 group also increased(P < 0.05). The mitochondrial membrane potential in the G3 group was higher than that of the control group (P < 0.05).Conclusion:Adding a certain concentration of citric acid in the cryoprotectant can reduce the damage of the mitochondria during the freezing and thawing.

Abstract:Objective:To investigate the effects and mechanisms of docosahexaenoie acid(DHA) with different concentrations on large conductance calcium-activated potassium channels (BK channels)in normal rat coronary smooth muscle cells (SMCs). Methods:Coronary SMCs were isolated by enzyme digestion. Effects of different concentrations of DHA on BK channels in absence and presence of cytochrome P450 (CYP) epoxygenase inhibitor SKF525A were studied by patch clamp in whole-cell configuration. Effects of DHA with different concentrations on BK single channels were studied by patch clamp in inside-out single channel configuration. Results:0.01-1.0 μmol/L(defined as low concentrations) of DHA activated whole-cell BK currents with a half effect concentration(EC50) of(0.24 ± 0.05)μmol/L. This effect could be abolished by pre-incubation with SKF525A. Concentration of 3-10 μmol/L (defined as high concentrations)of DHA activated whole-cell BK channels with an EC50 of(2.38 ± 0.22)μmol/L. However,this effect could not be abolished by pre-incubation with SKF525A. In the presence of 0,0.01,0.03,0.1,0.3,and 1 μmol/L DHA,the open probabilities of BK channels at 1 μmol/L calcium in external solution and test potentials at 60 mV were (0.095 2 ± 0.009 5),(0.093 9 ± 0.012 6),(0.098 6 ± 0.016 9),(0.099 5 ± 0.014 7),(0.097 5 ± 0.010 4),and(0.102 3 ± 0.020 6)respectively(P > 0.05,n=5). Low concentrations of DHA could not activate BK single channels. On the setting of 3 and 10 μmol/L DHA,the open probabilities of BK channels at 1 μmol/L calcium in external solution and test potentials at 60 mV were(0.700 3 ± 0.013 2),and(0.892 7 ± 0.053 2),respectively (P < 0.05,n=5). High concentrations of DHA could directly activate BK channels. Conclusion:BK channels can be activated by DHA in multiple mechanisms. Low concentration DHA can activate BK channel through CYP epoxygenase metabolites,while high concentration DHA can directly activate BK channel by binding to the channel proteins.

Abstract:Objective:To investigate the response of human pulmonary artery smooth muscle cells(PASMCs)to hypoxia with or without Xinmailong injection(XML),and explore the mechanism for XML to remit pulmonary artery hypertension(PAH). Methods:Na2S2O4 was used for building the hypoxic pulmonary artery smooth muscle cell model. The changes of apoptosis rate and apoptosis associated genes were detected by flow cytometry and realtime PCR,respectively,and membrane potentials and cytoplasmic Ca2+ under hypoxic stimulation were investigated by a laser scanning confocal microscope. Results:Under hypoxia condition,the proliferation of PASMCs was significant compared with that of the control group(P < 0.05),however,this effects can be markedly retarded by the treatment of XML(P < 0.05). Moreover,under hypoxia condition,Bcl-2 expression in PASMCs was obviously increased,while Bax expression was decreased when compared with those of the control group(P < 0.05). Noteworthily,XML can reverse both Bcl-2 and Bax expression induced by hypoxia(P < 0.05). In addition,XML stabilized the membrane potential and reduced the cytoplasmic Ca2+ under the hypoxic condition. Conclusion:Hypoxia leads to the proliferation of PASMCs,depolarization of membrane and increase of cytoplasmic Ca2+. The Chinese medicine XML can reduce all the effects induced by hypoxia,thereby our work might provide the experimental evidence for therapeutic strategy for pulmonary artery hypertension.

Abstract:Objective:To explore the scintigraphy of CGRRAGGSC labeled with 99mTc in nude mice bearing human breast cancer MCF-7. Methods: The CGRRAGGSC peptide was labeled with 99mTc using DTPA as a bifunctional chelator. Paper chromatography method was used to calculate the labeling rate and detect the stability. Immunofluorescence staining was used to evaluate the binding status of CGRRAGGSC peptide to human breast cancer cell. SPECT static scintigraphy at 0,0.5,1.0,2.0,3.0,4.0 h was performed in nude mice bearing human breast cancer after intratumoral injection of 1.85MBq/0.05 mL 99mTc-DTPA-CGRRAGGSC,99mTcO-4 and 99mTcO-4+DTPA-CGRRAGGSC,respectively. Immunohistochemistry assay was performed to observe the IL-11Rα protein levels in breast cancer. Results:99mTc-DTPA-CGRRAGGSC was radiolabeled successfully,the labeling rate was over 95% and the radiochemical purity in PBS and serum both were over 90% after 12 h at 37℃. Human breast cancer cells showed high binding abilities to CGRRAGGSC peptide. The scintigraphy showed that radioativity still deposited in tumor tissue at 4 h after intratumoral injection of 99mTc-DTPA-CGRRAGGSC,but radioactivity had faded obviously at 0.5 h after intratumoral injection of 99mTcO-4 or 99mTcO-4+DTPA-CGRRAGGSC. Breast cancer tissue had a higher expression of IL-11Rα. Conclusion:99mTc-DTPA-CGRRAGGSC can target the tumor tissue in nude mice bearing human breast cancer MCF-7,which lays a foundation for further study on the targeting therapeutic effect of therapeutic radionuclides rabeling CGRRAGGSC on breast cancer with expression of IL-11Rα.

Abstract:Objective:To explore the molecular mechanism in which IL-8 receptor promoted gastric cancer metastasis. Methods:Gastric cancer cells were cultured and transfected with IL-8 receptor antisense expression plasmid pcDNA3.1(+)/as-IL-8G and IL-8 receptor sense expression plasmid pcDNA3.1(+)/s-IL-8G. Protein expression was detected by Western blotting. Cell invasion was performed in Transwell and the invasion capacity was expressed by the relative invasive cell number. Results:Gastric cancer cells with lower expression of IL-8 receptor had significantly reduced invasion ability,and the expression of c-Jun,Ets-1 and MMP-9 decreased correspondingly,while higher expression of IL-8 receptor markedly enhanced the invasion capacity of gastric cancer cells,and the expression of c-Jun,Ets-1 and MMP-9 increased significantly. MMP-9 protein expression level decreased significantly after blocking the c-Jun or Ets-1 expression by phosphorylated modification antisense oligonucleotide technology,and the cell invasion ability decreased obviously at the same time. Conclusion:IL-8 receptor promoted gastric cancer cells metastasis through abnormal c-Jun and Ets-1-mediated MMP-9.

Abstract:Objective:To explore the effect of BANCR in reversing cisplatin(DDP) resistance of human non-small cell lung cancer(NSCLC)A549/DDP cell line and its related mechanisms. Methods:The real-time fluorescent quantitative PCR(qRT-PCR) was used to detect BANCR expression differences in DDP-resistant A549/DDP cells and its parent cell line A549.Overexpress BANCR in A549/DDP cells and use MTT assay,colony formation assay and flow-cytometric analysis to detect A549/DDP cells to DDP drug sensitivity,cell proliferation ability and apoptosis on treatment with DDP.The protein expression of p53 in A549/DDP cells were detected by western blotting. Results:The BANCR expression level in A549/DDP cell line was significantly lower than that in A549 cell line (P < 0.05). Ectopic BANCR expression in A549/DDP cells could produce the following effects:compared to the control group,the half inhibition concentration (IC50) of DDP in A549/DDP/BANCR cells lower (P < 0.05),proliferation of A549/DDP/BANCR cells diminished(P < 0.05),apoptosis on treatment with DDP increased(P < 0.05). Furthermore,the protein expressions p53 was upregulated in BANCR transfected group than those in empty vector. Conclusion:BANCR reverses the resistance of A549/DDP cells to DDP through induction of apoptosis,diminished cell proliferation,up-regulating the expressions of p53 protein.

Abstract:Objective:To prepare curcumin-loaded poly lactic-co-glycolic acid nanoparticles(Cur-PLGA-NPs),to characterize their physicochemical properties,and to study the in vitro release behavior and antitumor activity on human prostate cancer PC-3 cells in vitro. Methods:The Cur-PLGA-NPs were prepared by emulsification-solvent evaporation method. The transmission electron microscope was used to observe the particle appearance,Zetasizer instrument was used to detect the diameter,and ultracentrifugation was utilized to determine drug-loading rate and entrapment rate. Dynamic dialysis method was used to study the in vitro release behavior of Cur-PLGA-NPs. The antitumor activity on PC-3 cells was determined by CCK-8 method. Cells apoptosis and apoptosis rate were examined by using flow cytometry. Ultrastructural changes of cells were observed under the electronic microscopy,and compared to free Cur groups. Results:The optimal NPs were round with the nanometric size of(165.36 ± 24.21)nm,high entrapment rate of(83.05 ± 1.07)% and drug-loading rate of(10.87 ± 0.58)%. In vitro release test showed that release rate within 24 h of Cur-PLGA-NPs was 44.02%,and accumulating release rate was 84.81% at day10. After an effection of 5~40 μmol/L Cur and Cur-PLGA-NPs on PC-3 cells for 24~72 h,the inhibition rates of Cur and Cur-PLGA-NPs were 9.38%~83.62% and 10.56%~89.53%. CCK-8 experience revealed that there was significant difference between 20 μmol/L group and 40 μmol/L group compared Cur with Cur-PLGA-NPs(P < 0.05). Flow cytometry indicated that there was a significant difference between experiment group and control group(P < 0.05). Transmission electron microscopy showed that Cur-PLGA-NPs induced morphological change of apoptosis on PC-3 cells. Conclusion:The Cur-PLGA-NPs has good drug sustained-release capacity in vitro,and significantly improves the capacity of Cur to kill and inhibit the proliferation of PC-3 cells in vitro.

Abstract:Objective:Based on the traditional transformation methods of polyethylene glycol/lithium acetate,the membrane permeability of Candida albicans cell was increased by modifying chemical treatment method,and further to change the efficiency of plasmid transformation. Methods:The recombinant plasmid was linearied using restriction enzyme,and then the transformatiom efficiency was detected through adding different concentrations of dimethyl suifoxide(DMSO)(1%,5%,and 10%)or taking various length of heat shock(0.5 h,2 h,3 h and 4 h)based on the traditional transformation methods. The optimum concentration of DMSO and the best length of heat shock were determined according to the transformation efficiency experiment. Finally,we cultured the Candida albicans on the plates of SC select medium using these two methods to compare the plasmid transformation efficiency between these two groups. Results:The efficiency of plasmid transformation using the common method is 0.6×105 transformants/1 μg plasmid DNA/108 cells,while the efficiency of that using the improved method is 1.5×105 transformants/1 μg plasmid DNA/108 cells. The differences between those two groups were statistically significant. Conclusion:The improved method increases the Candida albicans plasmid transformation efficiency.

Abstract:Objective:To observe the protective effects of short-term(24 h)fasting pretreatment on hepatic ischemia/reperfusion injury patients undergoing excision of partial liver after occlusion of hepatic blood flow. Methods:A prospective randomly controlled study was conducted. Eighty patients with liver tumor classified as Child-Pugh class A undergoing hepatectomy were enrolled in the First Affiliated Hospital of NJMU from November 2015 to May 2016. The patients were randomized into control group and experimental group(each patient received a 24 hour-fasting,8 hour-fasting for water).Those patients then were divided into subgroups according to tumor size and time of clamping porta hepatis. Complete hepatic functions were determined before and after operation(1,3,7 d). Results:Compared with those indexes before operation,the postoperative levels of afalanine transaminase(ALT),aspartate transaminase (AST),total bilirubin (TB) and direct bilirubin (DB) in control and the experimental groups were both significantly elevated. ALT in the experimental group after operation(1,3,7 day)was significantly lower than that in control group(P < 0.05). Although TB and DB in the experimental group after operation(1,3 day) were lower than those in the control group(P < 0.05). No statistical significance were found in total protein,albumin and the rate of vomplication between the two groups(P > 0.05). It was significant that postoperative levels of ALT(1,3 day) in the experimental group were lower than those in the control group regardless of tumor size and time of clamping porta hepatis(P < 0.05). Conclusion:Hepatic I/R injury may occur in patients undergoing resection of tumor with occlusion of hepatic blood flow. Short-term fasting pretreatment may inhibit the release of serum pro-inflammatory cytokines and increase anti-inflammatory factor productions and the effects of autophagy,thereby alleviate hepatic I/R injury.

Abstract:Objective:Some adiponectin gene polymorphisms were associated with the risk of many inflammatory diseases. This study was designed to investigate the relationship between single nucleotide polymorphisms in adiponectin genes and hypertensive intracerebral hemorrhage in a Chinese population. Methods:SNaPshot was adopted to detect the adiponectin genotypes and allele distributions of rs2241766,rs1501299,rs17300539 and rs266729 in 150 patients with hypertensive intracerebral hemorrhage and 150 healthy controls. Results:The frequency distributions of GG genotype and G allele of rs266729 locus were statistically significantly different between the patients and the controls. The hypertensive intracerebral hemorrhage risk of GG genotype of rs266729 locus was 2.623 times higher than that of CC genotype (95% confidence interval,1.282-5.366;P=0.008). This difference retained significant using a logistic regression after multivariate adjustment for the other possible confounders. However,the risk of hypertensive intracerebral hemorrhage was not associated with rs2241766,rs1501299 and rs17300539 locus. Conclusion:Rs266729 gene polymorphism of adiponectin gene is related to hypertensive intracerebral hemorrhage. GG genotype and G allele of rs266729 locus may increase the risk of hypertensive cerebral hemorrhage in a Chinese population.

Abstract:Objective:To investigate the correlation between 6p21.1 rs2494938 and 7p15.3 rs2285947 and risk of cervical cancer. Methods:A total of 571 cervical cancer patients and 657 cancer-free controls were recruited in the study. The genotypes of 6p21.1 rs2494938 and 7p15.3 rs2285947 were detected by TaqMan MGB probe method. The correlation between the SNPs and the susceptibility to cervical cancer was evaluated using unconditional logistic regression analysis,and the relative risk of odds ratio(OR)and 95% confidence interval(CI) were calculated. Results:There was no significant association between the frequencies of GA or AA among the case and control groups at SNP rs2494938 (P=0.848). There was significant association between the frequencies of GA or AA among the case and control groups at SNP rs2285947,(P=0.028),and significant reduction was found in the risk of cervical cancer between all mutation genotype(GA + AA)and wild type genotype GG(OR=0.77,95%CI:0.62~0.97,P=0.025). Conclusion:There was no significant association between the rs2494938 pleiotropy and cervical cancer susceptibility,and there was significant association between the rs2285947 pleiotropy and cervical cancer susceptibility.

Abstract:Objective:To survey the prevalence of drug resistant and subtypes HIV in Jiangsu province in 2015. Methods:WHO truncated sequential sampling technique was adopted by using 47 samples of newly diagnosed as HIV positive and aged <25 years in Nanjing. RNA was extracted and HIV pol region was amplified by RT-PCR and nested PCR. Pol genetic mutation associated with transmitted drug resistance(TDR)was analyzed. Results:The success rate for sequence acquisition in the survey was 100%(47/47). The main subtype was CRF01_AE(59.6%),the rest subtypes in the sequence were CRF07_BC(25.5%),C(6.4%),B(2.1%),additional with 3 URFs. The prevalence of TDR in Jiangsu was low,and only 1 surveillance drug-resistance mutation(SDRMs)in protease inhibitor(PI)(M46I)was found by analyzing the 47 sequences. The individual with drug resistant HIV stain found in this study was infected by homosexual transmission and the subtype was CRF01_AE. The point finding in this study is that the percentage of HIV students was 46.8%(22/47),exceeding the percentage of HIV business servicers of 29.8%(14/47). A total of individuals with HIV were infected by homosexual transmission,except one by heterosexual. Conclusion:TDR has arisen in Jiangsu. Though the prevalence was low,we should still pay a closed attention on the tendency of TDR change. Students under age 25 are becoming the main constitution in newly diagnose HIV infections. Scanning up on consulting,testing and behavioral intervention are beneficial to HIV prevention and control. The key point on prevalence of TDR is to take preventive measures between men who have sex with men(MSMs).