Objective To assess the therapeutic effect of microwave radiation combined with Adriamycin liposome on liver VX2 tumor in rabbits. Methods VX2 tumor pieces were successfully implanted into liver lobes of rabbits with liver tumors formation. All rabbits were divided randomly into 4 groups (each n=8). Group A underwent transcatheter arterial chemoembolization (TACE) with Adriamycin, group B were treated with Adriamycin liposome, group C were treated with microwave radiation combinate with Adriamycin, and group D were treated with microwave radiation combinate with Adriamycin liposome. Spiral CT scans was performed to measure size of liver tumors at 1 day before operation and 14 days after operation, and liver function was tested at 1 day before operation, 1 day and 7 days after operation. Breathing frequency, heart rate and body temperture were measured before microwave radiation processing of microwave radiation, at 30 min after microwave radiation and 1 h after microwave radiation. All experimental animals were scarficed on the 15th day after operation and followed by pathologic and histologic examination of the tumor and after the operation with correlative comparisons. Results There was no significant difference in volumes and liver function among 4 groups at 1 day before operation. At 14 days after operation, the average tumor volume was (6.02±1.21) cm3 in group A, (5.74±1.43) cm3 in group B, (3.26±0.37) cm3 in group C and (1.89±0.14) cm3 in group D. There was no significant difference of liver function among 4 groups. After microwave radiation the rabbit's vital signs were stable. The intratumoral necrosis was more significant in group D than in the others (P<0.05). Conclusion Adriamycin liposome treatment for liver tumor via vascular interventional method combined with microwave radiation can significantly inhibit the growth of liver VX2 tumor in rabbits. Adriamycin liposome is a kind of effective pracputium in interventional chemoembelization, and the united therapy is safe and acceptable.

Objective:To study the immunoisolation effects of alginate-polylysine-alginate(APA)-microcapsules in vito.Methods:To observe changes of cellular immune and tectology of sheeps after hollow microcapsules, Bovine Chromaffin Cells(BCCs) and APA Microencapsulated BCCs(APA-BCCs) transplanted to the spinal subarachnoid space.Results:The blood lymphocyte numbers of each group didn't change; APA-microcapsule could prevent blood CD4 +T lymphocyte , CD4 +/CD8 + and CSF lymphocyte from increasing caused by BCCs transplantation; APA microencapsulation could reduce histological reactions in transplantation area and prolong the survival of the transplant.Conclusion:APA microcapsules possess the immunoisolation effects and can efficiently prevent immunoreactions.

Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKI) have shown great efficacy in the treatment of non-small cell lung cancer (NSCLC) patients with EGFR-mutation positive tumors.However,the response to EGFR-TKI is quite different even in EGFR-mutation positive patients.Besides that,different lesions in same patient can also show different response to EGFR-TKI.These phenomena might be associated with the heterogeneity of EGFR mutations,which involves intratumoral heterogeneity,intertumoral heterogeneity,and the heterogeneity before and after treatment.The article introduces the advance in heterogeneity of EGFR mutations from these three aspects.

Objective To investigate the role of dendritic cell (DC) subsets and T cell subpopulations expressing different intracellular cytokines in the pathogenesis of syphilis. Methods Flow cytometry was utilized to measure the frequencies of peripheral DC subsets and IFN-γ-, IL-2-, IL-4-, IL-10-expressing peripheral CD4+ and CD8+ T cell subpopulations in 20 patients with syphilis before and after therapy, and in 15 healthy human controls. Results The frequency of CD11c+ DC was significantly lower in pretreated patients than that in treated patients (P<0.01), but higher than that in the normal controls (P<0.05), and the same was true for the frequency of IFN-γ-expressing T cells, IL-2-expressing T cells, and IL-12-expressing T cells. By contrast, a decrease was noticed in the frequency of IL-4-expressing T cells and IL-10-expressing T cells in pretreated patients compared with that in the normal controls. Furthermore, in patients with syphilis, the frequency of CD11c+ and CD123+ DC was unrelated to the percentage of IFN-γ-expressing and IL-12- expressing T cells (both P>0.05). Conclusions IFN-γ, IL-2 and IL-12 may exert an immunoregnlatory effect in the eliminating of Treponema pallidum, whereas IL-4 and IL-10 may perform an imrnunoinhibitory effect.

AIM To invesigate the effect of subarachnoid transplantation of APA microcapsulized bovine chromaffin cells (BCCs) on mRNA expression for Nav1.8 in the dorsal root ganglia neurons(DRG) of rats with neuropathic pain by means of in situ hybridization. METHODS SD rats were randomly divided into four groups of five. Normal rats were used as control group (group C). Rats with right sciatic nerve been ligated were used as CCI group. Five to six hundred empty APA microcapsules(group APA) or 5?10 6 APA microcapsulized BCCs (group APA-BCCs) were grated into subarachnoid space of CCI rats 7 days after operation. Allodynia and hyperalgesia were measured by Von-Frey filaments and CO 2 laser 7 days after transplantation. DRG in lumbar four and five was taken out and 15 ?m freezing sections were made 7 days after tansplantation. Sections was used to detect mRNA expression for TTX-resistent Na + Nav1.8 by in situ hybridization with Dig-labeled RNA probe. RESULTS The mRNA hybridization signal for Nav1.8 in DRG of group CCI and group APA was lower than that of group C. The expression of mRNA for Nav1.8 in DRG was higher in group APA-BCCs than that in group CCI and group APA with abatement of allodynia and hyperalgesia. There was no difference in the mRNA hybridization signal for Nav1.8 in DRG between group APA-BCCs and group C. CONCLUSION mRNA expression for Nav1.8 in DRG of CCI ratswas down-regulated. APA microcapsulized BCCs grafting can reverse the down-regulation of mRNA expression for Nav1.8 in DRG of CCI rats. Restoration of mRNA expression for Nav1.8 in DRG contributes to the analgesic effect of subarachnoid transplantation of APA microcapsulized BCCs.

Objective To observe the effect of APA-BCC(alginate-polylysine-alginate microencapsulated bovine adrenal medullary chromaffin cells)microcapsules transplantation into the subarachnoid space of cancer pain patients on endogenous opioid peptides and catecholamine concentration in cerebrospinal fluid(CSF).Methods The different doses of APA-BCC microcapsules were transplanted into the spinal subarachnoid space of cancer patients with moderate or serious pain.The concentrations of Leu-enkephalin(L-EK),β-endorphin(β-EP),dynorphin A(DynA),noradrenaline(NA)and adrenaline(AD)in CSF were tested.Results L-EK concentration in CSF increased remarkably after transplantation,and the increase was most remarkable when transplantation doses were 1.0×107and 1.25×107;there were no remarkable changes of β-EP,DynA,NA and AD after transplantation.Conclusion The analgetic effects of APA-BCC microcapsules tranplantation may correlate with the increase of L-EK in CSF of cancer pain patients;the dose of 1.0×107 and 1.25×107cells may be the most effective dose.

Objective:To establish a method based on the iPLEX analysis of MassARRAY mass spectrometry platform to detect multiplex genetic mutations among Chinese lung cancer patients. Methods:We reviewed the related literature and data of lung cancer treatments. We also determined 99 mutation hot spots in 13 target genes, namely, EGFR, KRAS, ALK, FGFR1, FGFR2, FGFR3, PIK3CA, BRAF, PTEN, MET, ERBB2, AKT1, and STK11, which are closely related to the pathogenesis, drug resistance, and metastasis of lung cancer and are associated with relevant transduction pathways. A total of 297 primers comprising 99 paired forward and reverse amplification primers and 99 matched extension primers were designed by using Assay Design in accordance with the mutation label and format requirements of the MassARRAY platform. The detection method was established by analyzing eight cell lines and six lung cancer specimens;the proposed method was then validated through comparisons with a LungCarta kit. The sensitivity and specificity of the proposed method were evaluated by directly sequencing EGFR and KRAS genes in 100 lung cancer cases. Results:The proposed method could detect multiplex genetic mutations in the lung cancer cell lines, and this finding is consistent with that observed using previously reported methods. The proposed method could also detect such mutations in clinical lung cancer specimens;this result is also consistent with that observed by using the LungCarta kit. However, an FGFR2 mutation was detected only by using the proposed method. The measured sensitivity and specificity were 100%and 96.3%, respectively. Conclusion:The proposed MassARRAY technology-based method could detect multiplex genetic mutations among Chinese lung cancer patients. Indeed, the proposed method can be potentially applied to detect mutations in cancer cells.

Objective To establish a HRM assay to screen for KRAS mutations in clinical colorectal cancer patients.Methods The sensitivity of HRM was analyzed by detecting somatic mutations in exon 2,notably codons 12 and 13 of the KRAS gene in the serial plasmid mixture samples which were mixed using the different proportions mutation plasmid and wide type plasmid of KRAS.HRM analysis was performed for KRAS on DNA insolated from a panel of 60 colorectal cancer samples derived from fresh tissues.The results were compared with the direct sequencing data.Results After the PCR amplification,the mutation results could be available by performing HRM analysis in the same tube on a real time PCR machine with HRM capability.HRM detection could identify KRAS mutation in a proportion of 10% of mutation plasmid DNA.All 60 samples identified the KRAS mutation by HRM and sequencing.17 samples were positive(28.3%) by HRM for KRAS exon 2 mutations,and 15 samples were confirmed the presence of codon 12 or 13 mutations(25.0%) and the other 2 samples were wild type by sequencing.The 60 samples detected by HRM were given 100% sensitivity with 96% specificity.Conclusions HRM is a sensitive intube methodology to screen for mutations in clinical samples.HRM will enable high-throughput screening to gene mutations to allow appropriate therapeutic choices for patients and accelerate research aimed at identifying novel mutations in human cancer.

Objective:To explore the effect of hydrogen sulfide (H 2S) on modulating the subunit Kir6.2 of adenosine triphosphate sensitive potassium channels via the cyclic guanosine monophosphate-dependent protein kinase (cGMP/PKG) signaling pathway in epileptic rat models. Methods:Sixty adult male SD rats were randomly divided into the following six groups (10 rats in each group) by random number table method: control, epileptic, H 2S donor, H 2S donor+epileptic, KT5823 (one inhibitor of the cyclic guanosine monophosphate-dependent protein kinase)+H 2S donor+epileptic, and glibenclamide (one inhibitor of the adenosine triphosphate sensitive potassium channels)+H 2S donor+epileptic groups. Except the control group, SD rats were intraperitoneally injected with plentylenetetrazole to make the kindling models and their behaviours were recorded including the latency period, the grade, and the duration of the first epileptic seizure according to the Racine′s standard. The waveforms of electroencephalogram (EEG) in hippocampus were also recorded during the seizure. The mRNA and protein levels of PKG and Kir6.2 in hippocampus were evaluated by Western blotting and quantitative real-time polymerase chain reaction, and the hippocampal concentrations of cGMP and phosphorylation of cyclic guanosine monophosphate-dependent protein kinase (p-PKG) were detected by enzyme linked immunosorbent assay. Results:Rats in the epileptic group showed Ⅳ-Ⅴ grade of epileptic seizure [4.500 (4.000, 4.875)], short latency period [(10.37±8.21) min] but long duration [(69.50±24.37) s] of seizure. Compared to the epileptic group, rats in the H 2S donor group showed Ⅱ-Ⅲ grade of epileptic seizure ( P=0.004), significantly longer latency period ( P<0.001), and shorter duration of seizure ( P<0.001). Compared to the H 2S donor+epileptic group, rats in the KT5823+H 2S donor+epileptic group showed Ⅲ-Ⅳ grade of epileptic seizures, significantly shorter latency period ( P<0.001), and longer duration of seizure ( P<0.001). The results of EEG showed that the wave patterns in the epileptic group were spike or sharp waves and the amplitudes were largest [(190.570±23.590) μV]. Compared with the epileptic group, amplitudes were reduced ( P<0.001) in the H 2S donor+epileptic group. PKG mRNA and PKG protein were expressed differently among all groups (PKG mRNA: n=5, H=26.714, P<0.001; PKG protein: n=5, F=30.597, P<0.001). Compared with the control group, the expression of both PKG mRNA and PKG protein was decreased (PKG mRNA: 1.000±0.001 vs 0.782±0.064, P=0.023; PKG protein: 0.550±0.037 vs 0.145±0.020, P=0.042) in the epileptic group. Besides, Kir6.2 mRNA and Kir6.2 protein were expressed differently among all groups (Kir6.2 mRNA: n=5, H=27.761, P<0.001; Kir6.2 protein: n=5, F=60.659, P<0.001). Compared with the control group, the expression of both Kir6.2 mRNA and Kir6.2 protein was decreased (Kir6.2 mRNA: 1.000±0.001 vs 0.897±0.033, P=0.004; Kir6.2 protein: 0.384±0.035 vs 0.215±0.016, P=0.024) in the epileptic group. And the concentrations of cGMP and p-PKG were decreased (cGMP: P<0.001; p-PKG: P<0.001) in the epileptic group. The results in the H 2S donor+epileptic group were up-regulated (PKG mRNA: P=0.047; PKG protein: P<0.001; Kir6.2 mRNA: P=0.011; Kir6.2 protein: P<0.001; cGMP: P<0.001; p-PKG: P<0.001) compared with the epileptic group. However, the results in the KT5823+H 2S donor+epileptic group were down-regulated (PKG mRNA: P=0.015; PKG protein: P=0.027; Kir6.2 mRNA: P=0.013; Kir6.2 protein: P=0.017; cGMP: P=0.005; p-PKG: P<0.001) compared with the H 2S donor+epileptic group. Conclusion:A possible mechanism is that H 2S prevents the epileptic seizure from modulating the subunit Kir6.2 of ATP sensitive potassium channels via the cGMP/PKG signaling pathway.

ObjectiveTo investigate the fusion sequence complexity of EML4-ALK in non-small cell lung cancer (NSCLC) patients,and the potential mutation in tyrosine kinase ( TK ) domain of ALK gene.MethodsIn routine practice,a novel echinoderm microtubule-associated protein-like4 and anaplastic lymphoma kinase (EML4-ALK) V3c variant was detected by rapid amplification of cDNA ends-polymerase chain reaction ( RACE-PCR )-sequencing technology in a patient with NSCLC.The further consecutive 39 cases( total of 40 cases)were screened by use of reverse transcription (RT)-PCR for EML4-ALK fusion.Positive PCR products were purified and cloned into T vectors,transformed into DH5a germ cells and colony picked up and sequenced for sequence complexity analysis.Tyrosine kinase domain of ALK was amplified by RT-PCR and sequenced.ResultsThree out of 40 cases had EML4-ALK fusion.One case had six novel variants of EML4-ALK co-existing,termed as V3c ( 64.6％ ),V3d ( 25.0％ ),V3e ( 2.1％ ),V3f (4.2％ ),V3g(2.1％ )and V3h(2.1％ ) variants,whereas without common V3a and V3b variants.In other two positive cases,one was V1 variant,another was concurrent V2,V3a and V3b variants.No mutations were detected in the TK domain of EML4-ALK in any case.ConclusionsSeveral EML-ALK variants could co-exist in a given lung cancer tissue,which suggest that the diversity and sequence complexity of EML4-ALK fusion are exist.Attentions should be paid to screen all the variants in clinic to improve the pick-up rate.