[Allergic bronchopulmonary aspergillosis with low serum IgE: a case report].

Here, we reported a case of delayed diagnosis of allergic bronchopulmonary aspergillosis (ABPA) with low serum IgE and normal Aspergillus fumigatus-specific IgE levels. During the course of the disease, the patient (female, 55 years old) had imaging manifestation of mass shadow and significant elevation of carcinoembryonic antigen, leading to suspicion of a lung tumor. Later, transbronchial lung biopsy tissue culture showed Aspergillus fumigatus. Combined with the history, clinical characteristics and imaging, she was diagnosed with allergic bronchopulmonary aspergillosis combined with invasive pulmonary aspergillosis. As the diagnostic criteria for ABPA do not cover all patients with ABPA, in rare cases where immunological evidence is insufficient, a combination of clinical and imaging features is required for early diagnosis and treatment.

[Prevalence betel nut abuse and its related factors among residents in Haikou City from 2013 to 2014].

From 2013 to 2014, the age of 924 residents recruited in Haikou City was (38±13) years old, of which 57.3% (529) were males. Those who chewed betel nuts accounted for 17.4% (161). According to the diagnostic criteria of substance abuse in the Fourth Edition of the Diagnostic and Statistical Manual of Mental Disorders, the detection rate of betel nut abuse was 7.0% (65). Compared with those who were>30 years old, educated>6 years and non-smokers, people aged 15 to 30 years, education level less than 6 years and smoking behavior had higher risk of betel nut abuse, with the OR (95%CI) about 4.21 (1.48-11.99), 7.81 (1.92-31.69), and 13.53 (4.15-44.11), respectively.

Molecular characterization of prophenoloxidase-1 (PPO1) and the inhibitory effect of kojic acid on phenoloxidase (PO) activity and on the development of Zeugodacus tau (Walker) (Diptera: Tephritidae).

Phenoloxidase (PO) plays a key role in melanin biosynthesis during insect development. Here, we isolated the 2310-bp full-length cDNA of PPO1 from Zeugodacus tau, a destructive horticultural pest. qRT-polymerase chain reaction showed that the ZtPPO1 transcripts were highly expressed during larval-prepupal transition and in the haemolymph. When the larvae were fed a 1.66% kojic acid (KA)-containing diet, the levels of the ZtPPO1 transcripts significantly increased by 2.79- and 3.39-fold in the whole larvae and cuticles, respectively, while the corresponding PO activity was significantly reduced; in addition, the larval and pupal durations were significantly prolonged; pupal weights were lowered; and abnormal phenotypes were observed. An in vitro inhibition experiment indicated that KA was an effective competitive inhibitor of PO in Z. tau. Additionally, the functional analysis showed that 20E could significantly up-regulate the expression of ZtPPO1, induce lower pupal weight, and advance pupation. Knockdown of the ZtPPO1 gene by RNAi significantly decreased mRNA levels after 24 h and led to low pupation rates and incomplete pupae with abnormal phenotypes during the larval-pupal interim period. These results proved that PO is important for the normal growth of Z. tau and that KA can disrupt the development of this pest insect.

[Genotyping by CRISPR and regional distribution of Yersinia pestis in Qinghai-plateau from 1954 to 2011].

Objective: To investigate the CRISPR genotypes (clusters) and regional distribution of Yersinia pestis in Qinghai-plateau. Methods: One hundred and two isolates of Y. pestis isolated from human plague patients, host animal and insect vectors from Qinghai-plateau were selected. The DNAs were extracted using the traditional sodium dodecyl sulfate decomposition and phenol-chloroform method. Three CRISPR loci YPa, YPb and YPc of 102 isolates of Y. pesits were amplified and sequenced, and then the CRISPR sequence analysis was carried out by comparing the latest published CRISPR spacer dictionary and the NCBI database to identify the spacer and spacer array. CRISPR genotyping of isolates of Y. pesits were finally conducted according to the polymorphism of the spacer arrays and the regional distribution pattern of isolates of Y. pesits in Qinghai-plateau was described. Results: Forty spacers including 22 of YPa, 13 of YPb and 5 of YPc were observed among 102 isolates of Y. pestis in Qinghai-plateau, of which 5 spacers (a1', a103, a104, b4'' and b4''') were firstly identified. Meanwhile, 16, 10, and 5 different spacer arrays were obtained in YPa, YPb and YPc respectively, including 11 new spacer arrays detected in this study. One hundred and two isolates were divided into 24 CRISPR genotypes and classified into 9 CRISPR clusters (Cb4, Cb4', Cb2, Ca37, Ca7, Ca7', CaΔ5', Ca35' and Cc3'). Each dominant cluster presented significant aggregation geographically: Ca7 were found in Yushu, Nangqian, Chenduo, Zaduo, Zhiduo and Qumalai countries. Ca7' were found in Xunhua, Tongren, Zeku, Tongde, Maqin and Guinan countries. CaΔ5' were restricted to Qilian, Gangcha, Menyuan and Datong countries. CaΔ35' were found in Huangyuan, Haiyan, Gangcha, Tianjun, Delingha, Wulan, Doulan, Gonghe, Xinghai, Guide and Tongde countries. Conclusion: CRISPR-based genotyping analyses showed complicated population of Y. pestis in Qinghai-plateau. Four clusters, Ca7, Ca7', CaΔ5' and Ca35' were the most epidemic dominant four clusters and presented obvious regional distribution patterns, which instructed us to strengthen the surveillance and prevention and control by CRISPR-genotyping technique.

[Multiregion sequencing reveals intratumor heterogeneity in esophageal squamous cell carcinoma].

OBJECTIVE: To study the intratumor heterogeneity of esophageal squamous cell carcinoma (ESCC). METHODS: We used whole-exome sequencing and array-based comparative genomic hybridization to profile mutations and changes in copy number from 11 regions within 2 cases of ESCC and from the metastatic lymph nodes. RESULTS: The numbers of somatic single nuclear polymorphisms (SNPs) in 4 regions within the tumors in case 1 and case 2 were 93±17 and 124±28, respectively. The majority of SNPs were non synonymous mutations, synonymous mutations, nonsense mutations and splicing junction mutations. The average indels in the 4 tumor regions of case 1 and case 2 were 40±6 and 51±3, respectively. These small indels mainly occurred in exonic (frame-shift and non-frame-shift), untranslational regions of genes and splicing junction regions. All regions from a tumor exhibitedo bvious heterogeneity, and mutational similarity of all four regions within a tumor was less than 25%. Furthermore, gene copy number alteration (gain or loss) varied among multiple regions of a tumor, and the similarity of gene copy number was less than 20%. Phylogenetic analysis of the somatic mutation frequency suggests that multiple, genomic heterogeneous clones co-exist within a primary ESCC, and metastatic subclones may evolve from the primary non-metastatic parental clone. These results indicated that a single-region sampling can not reflect the architecture of the genomic landscape of mutations in ESCC tumors. CONCLUSIONS: Sequence analysis of whole genome exon in multiple regions can provide strong evidence for genomic heterogeneity in esophageal squamous cell carcinoma.

Antitumor effects of oncolytic adenovirus armed with PSA-IZ-CD40L fusion gene against prostate cancer.

Advanced prostate cancer (PC) still remains incurable. Novel immunogene therapy shows promise as treatment strategy that can target both localized and metastasized PC. In this study, we have developed a PC-specific oncolytic adenovirus (Ad-PL-PPT-E1A) armed with fusion gene of prostate-specific antigen and CD40 ligand, and aimed to evaluate its therapeutic effect in vitro and in vivo. After they were rescued in human embryonic kidney 293 cells, we confirmed that Ad-PL-PPT-E1A could mediate the expression of E1A efficiently and produce abundant progeny viruses in PC cells in vitro. Our data showed that Ad-PL-PPT-E1A induced apoptosis and resulted in specific oncolytic toxicity in PC cells, which was detected by Annexin-V staining and crystal violet, respectively. After stimulation with lysates, immune phenotypes and cytokines expression of human dendritic cells was detected by flow cytometry and real-time polymerase chain reaction, respectively. And, the results showed that the lysate of Ad-PL-PPT-E1A-infected LNCaP cells upregulated the expression of CD80, CD83, CD86 and mRNA level of interleukin-6 (IL-6), IL-12, IL-23 and tumor necrosis factor-α significantly. In established PC3M cell-xenografted mouse models, Ad-PL-PPT-E1A treatment improved the survival and suppressed the tumor growth obviously. In conclusion, Ad-PL-PPT-E1A exhibited enhanced antitumor activity is a promising approach for gene therapy of advanced PC.

Record-high and robust 17.125 Gb/s gross-rate over 25 km SSMF transmissions of real-time dual-band optical OFDM signals directly modulated by 1 GHz RSOAs.

Aggregated 17.125 Gb/s real-time end-to-end dual-band optical OFDM (OOFDM) transmissions over 25 km SSMF IMDD systems with 7 dB receiver sensitivity improvements are experimentally demonstrated, for the first time, by utilizing low-cost transceiver components such as directly modulated 1GHz RSOAs and DACs/ADCs operating at sampling speeds as low as 4GS/s. The demonstrated OOFDM transceivers have both strong adaptability and sufficiently large passband carrier frequency tunability, which enable full use of highly dynamic spectral characteristics of the transmission systems. This results in the achievements of not only excellent performance robustness to variations in system operating conditions but also significantly relaxed requirements on RSOA small-signal modulation bandwidth. It is shown that the aforementioned transmission capacity only varies by <23% over a RSOA-injected optical power variation range as large as 20dB, and that the 1 GHz RSOAs can support successful transmissions of adaptively modulated OOFDM signals having bandwidths of 8.5 GHz. By taking into account the adopted 25% cyclic prefix and a typical 7.3% FEC overhead, the demonstrated real-time OOFDM transmission systems are capable of conveying 11.6 Gb/s user data.

Robust real-time 15.125 Gb/s adaptive optical OFDM transmissions over 100 m OM2 MMFs utilizing directly modulated VCSELs subject to optical injection locking.

Optical injection locking (OIL) is an effective approach for significantly enhancing the modulation bandwidths of VCSELs. The frequency responses of OIL-VCSELs are, however, very sensitive to the applied OIL conditions. This brings about strong difficulties in practically utilizing the OIL-enhanced modulation bandwidths to achieve highly robust transmission performances of directly modulated OIL-VCSEL-based multi-mode fibre (MMF) links for cost-sensitive application scenarios such as data-centers. In this paper, directly modulated OIL-VCSEL-based real-time dual-band optical OFDM (OOFDM) transceivers with tunability in both the electrical and optical domains are experimentally demonstrated, for the first time, utilizing DACs/ADCs at sampling speeds as low as 4GS/s. The transceivers can support 15.125 Gb/s adaptive OOFDM transmissions over 100 m OM2 MMF links based on intensity modulation and direct detection. More importantly, the adaptability and tunability of the demonstrated transceivers enable the achievement of excellent robustness of the aggregated OOFDM transmission capacity to OIL condition variations. It is shown that, over a large diversity of OIL conditions that give rise to significantly different system frequency responses, the aggregated OOFDM transmission capacity only vary by <11% in the aforementioned transmission link.

Experimental demonstrations of record high REAM intensity modulator-enabled 19.25Gb/s real-time end-to-end dual-band optical OFDM colorless transmissions over 25km SSMF IMDD systems.

Record-high 19.25Gb/s real-time end-to-end dual-band optical OFDM (OOFDM) colorless transmissions across the entire C-band are experimentally demonstrated, for the first time, in reflective electro-absorption modulator (REAM)-based 25km standard SMF systems using intensity modulation and direct detection. Adaptively modulated baseband (0-2GHz) and passband (6.125 ± 2GHz) OFDM RF sub-bands, supporting signal line rates of 9.75Gb/s and 9.5Gb/s respectively, are independently generated and detected with FPGA-based DSP clocked at only 100MHz as well as DACs/ADCs operating at sampling speeds as low as 4GS/s. The two OFDM sub-bands are electrically multiplexed for intensity modulation of a single optical carrier by an 8GHz REAM. The REAM colorlessness is experimentally characterized, based on which optimum REAM operating conditions are identified. To maximize and balance the signal transmission performance of each sub-band, on-line adaptive transceiver optimization functions and live performance monitoring are fully exploited to optimize key OOFDM transceiver and system parameters. For different wavelengths within the C-band, corresponding minimum received optical powers at the FEC limit vary in a range of <0.5dB and bit error rate performances for both baseband and passband signals are almost identical. Furthermore, detailed investigations are also undertaken of the maximum aggregated signal line rate sensitivity to electrical sub-band power variation. It is shown that the aforementioned system has approximately 3dB tolerance to RF sub-band power variation.

Retrieving quasi-phase-matching structure with discrete layer-peeling method.

An approach to reconstruct a quasi-phase-matching grating by using a discrete layer-peeling algorithm is presented. Experimentally measured output spectra of Solc-type filters, based on uniform and chirped QPM structures, are used in the discrete layer-peeling algorithm. The reconstructed QPM structures are in agreement with the exact structures used in the experiment and the method is verified to be accurate and efficient in quality inspection on quasi-phase-matching grating.

A Novel Graphene Metal Semi-Insulator Semiconductor Transistor and Its New Super-Low Power Mechanism.

The state-of-art Si Matel-Oxide-Semiconductor Field-Effect-Transistor (MOS-FET) meets the problem of the Power Consumption (PC) can not be effecively deceased guided by the Moore's Law as before. The GFET has the problem of the device can not be effectively turned off, since the band-gap of the graphene is zero. To solve these problems, noticing the amount of the carriers in the 2 dementional semiconductor material is limited, we propose a Matel-Semi-Insulator-Semiconductor Field-Effect-Transistor (MSIS-FET) to replace the traditional MOS-FET. We verify our idea by fabricating the graphene MSIS-FETs using the natural Aluminium-oxide (Al-oxide) as the semi-insulator gate dielectric. From MSIS-FETs fabricated, we obtain following experimental results. The graphene MSIS-FET is turned off very well, a recorded high Ids on/off ratio of 5 × 107 is achieved. A saddle and close-loop shape transfer feature of Ids-Vgs is obtained first time for transistors. A non-volatile memory characteristics is observed. A carrier re-injection principle and a super-Low PC mechanism for semiconductor devices and integrated circuits (ICs) are found from the transfer feature of the graphene MSIS-FET. It is shown that the PC of the semiconductor devices and (ICs) can be reduced by over three orders of magnitude by using this new mechanism.

ICOS-Dependent and -independent functions of memory CD4 T cells in allograft rejection.

Donor-reactive memory T cells undermine the survival of transplanted organs through multiple pathways. We have previously reported that memory CD4 T cells resist treatment with anti-CD154 antibody and donor-specific transfusion (DST/MR1) and promote cardiac allograft rejection via generation of effector CD4 T cells and alloantibody. We hypothesized that the helper functions of memory CD4 T cells are independent of T-cell costimulation through CD154 but instead are regulated by alternative costimulatory pathways. This study investigated how blocking ICOS/B7RP-1 interactions affects functions of donor-reactive memory CD4 T cells. Treatment with blocking anti-ICOS mAb synergized with DST/MR1 and prolonged mouse cardiac allograft survival despite the presence of donor-reactive memory CD4 T cells. While blocking ICOS did not diminish the expansion of preexisting memory CD4 T cells or the induction of allospecific effector T cells, it did inhibit recruitment of the activated memory and effector T cells into the graft. In addition, anti-ICOS mAb treatment in combination with DST/MR1 prevented help provided by memory CD4 T cells for production of donor-specific IgG antibody. These results demonstrate the potential efficacy of ICOS blockade in sensitized transplant patients and provide the foundation for rational use of ICOS blockade in combination with other graft-prolonging strategies.

The cloning and expression of alpha-tubulin in Monochamus alternatus.

The Japanese pine sawyer Monochamus alternatus is one of the major forest pests. It damages pine directly and transfers the nematode Bursaphelenchus xylophilus to pine wood; resulting in serious economic losses around the world every year. Alpha-tubulin is one of most important proteins in most species. We cloned a ubiquitously expressed M. alternatus alpha-tubulin gene and analysed its nucleotides and protein structure; its sequence characters are consistent with what have been reported in other insects. The alignment of proteins showed that there is high homology of alpha-tubulin between M. alternatus and other species. Western blot and immunocytochemistry analyses suggested a common epitope of alpha-tubulin between M. alternatus and Strongylcentrotus purpuratus. We also expressed the protein in Escherichia coli for further functional studies.

Simultaneous determination of nucleobases, nucleosides and saponins in Panax notoginseng using multiple columns high performance liquid chromatography.

A new multiple columns HPLC method for simultaneous determination of 16 characteristic components, 5 nucleobases and nucleosides (uracil, cytidine, uridine, guanosine and adenosine), and 11 saponins (notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, notoginsenoside R4, notoginsenoside Fa, ginsenoside Rb1, notoginsenoside R2, ginsenoside Rg2, ginsenoside Rh1, ginsenoside Rd and notoginsenoside K), in the root of Panax notoginseng, a valued traditional Chinese medicinal herb, were developed. Notoginsenoside R4, Fa and K were first quantitatively determined in P. notoginseng. The 5 nucleobases and nucleosides compounds were separated on a Zorbax SB-Aq column (150 x 4.6 mm, 5.0 microm) and 11 saponins were analyzed using a Zorbax Bonus-RP column (150 x 4.6 mm, 5.0 microm) with column switching. The column temperature was set at 30 degrees C. Mobile phase was composed of 5mM ammonium acetate aqueous (A), water (B) and acetonitrile (C) using a gradient elution. The flow rate was 1.5 mL/min and detection wavelengths were set at 260 nm for nucleobases and nucleosides, and 203 nm for saponins. The developed method had good repeatability and sensitivity for quantification of 16 analytes with overall precision (including intra- and inter-day) less than 3% (RSD), and LOD and LOQ were less than 1.33 microg/mL and 5.12 microg/mL, respectively. The method was successfully applied to the simultaneous determination of 16 analytes in 15 samples of P. notoginseng collected from different places of China, which indicated that multiple columns HPLC can be used for comprehensive quality control of P. notoginseng.

A rapid method for simultaneous determination of 15 flavonoids in Epimedium using pressurized liquid extraction and ultra-performance liquid chromatography.

Herba Epimedii (family Berberidaceae), Yinyanghuo in Chinese, is one of commonly used Chinese medicines. Flavonoids are considered as its active components. In this study, a rapid ultra-performance liquid chromatography (UPLC) method was developed for simultaneous determination of 15 flavonoids, including hexandraside E, kaempferol-3-O-rhamnoside, hexandraside F, epimedin A, epimedin B, epimedin C, icariin, epimedoside C, baohuoside II, caohuoside C, baohuoside VII, sagittatoside A, sagittatoside B, 2''-O-rhamnosyl icariside II and baohuoside I in different species of Epimedium. The analysis was performed on Waters Acquity UPLC system with an Acquity UPLC BEH C18 column (50 mm x 2.1mm I.D., 1.7 microm) and gradient elution of 50mM acetic acid aqueous solution and acetonitrile within 12 min. All calibration curves showed good linearity (R2>0.9997) within test ranges. The LOD and LOQ were lower than 0.13 and 0.52 ng on column, respectively. The R.S.D.s for intra- and inter-day of 15 analytes were less than 5.0% at three levels, and the recoveries were 95.0-103.7%. The validated method was successfully applied to quantitatively analyze 15 flavonoids in different species of Epimedium. The results showed there were great variations among the contents of investigated flavonoids. Hierarchical clustering analysis based on characteristics of 15 investigated compounds peaks in UPLC profiles showed that 37 samples were divided into 3 main clusters, which were in accordance with their flavonoids contents. The simulative mean chromatogram of the high content cluster was generated to compare the samples from different species and/or locations of Epimedium. Four flavonoids including epimedin A, B, C and icariin were selected as markers for quality control of the species of Epimedium used as Yinyanghuo.

Simultaneous determination of 15 flavonoids in Epimedium using pressurized liquid extraction and high-performance liquid chromatography.

Herba Epimedii (family Berberidaceae), Yinyanghuo in Chinese, is one of the commonly used Chinese medicines. Flavonoids are considered as its active components. In this study, a reliable pressurized liquid extraction (PLE) and HPLC method was developed for simultaneous determination of 15 flavonoids, namely hexandraside E, kaempferol-3-O-rhamnoside, hexandraside F, epimedin A, epimedin B, epimedin C, icariin, epimedoside C, baohuoside II, caohuoside C, baohuoside VII, sagittatoside A, sagittatoside B, 2''-O-rhamnosyl icariside II and baohuoside I in different species of Epimedium. The analysis was performed by using a Zorbax SB-C18 analytical column (250 mm x 4.6 mm I.D., 5 microm) at gradient elution of water and acetonitrile with diode-array detection (270 nm). All calibration curves showed good linearity (r(2)>0.9997) within test ranges. The LOD and LOQ were lower than 1.31 ng and 2.62 ng on column, respectively. The RSD for intra- and inter-day of 15 analytes was less than 3.8% at three levels, and the recoveries were 90.5-106.8%. The validated method was successfully applied for the analysis of 15 flavonoids in different species of Epimedium which had great variation on the contents of investigated flavonoids. Hierarchical clustering analysis based on the characteristics of 15 investigated compound peaks in HPLC profiles showed that 26 samples were divided into three main clusters, which were in accordance with their flavonoid contents. Four flavonoids including epimedin A, B, C and icariin were optimized as markers for quality control of the species of Epimedium used as Yinyanghuo.

Optimization and comparison of three methods for extraction of volatile compounds from Cyperus rotundus evaluated by gas chromatography-mass spectrometry.

The essential oil of Cyperus rotundus has multiple pharmacological activities. Therefore, the extraction with high yield and quality is very important for preparation of essential oil of C. rotundus. In this paper, three methods, namely hydrodistillation (HD), pressurized liquid extraction (PLE) and supercritical fluid extraction (SFE), for extraction of volatile compounds from C. rotundus were optimized and compared by gas chromatography-mass spectrometry. Among eight identified compounds in C. rotundus, five components including alpha-copaene, cyperene, beta-selinene, beta-cyperone and alpha-cyperone were quantitatively determined or estimated using alpha-cyperone as standard, which showed that PLE had the highest extraction efficiency, while SFE had the best selectivity for extraction of beta-cyperone and alpha-cyperone. The contents of ingredients from C. rotundus extracted with HD, PLE and SFE are significantly different, which suggest that comparison of chemical components and pharmacological activities of different extracts is helpful to elucidate the active components in C. rotundus and control its quality.

Fractionated dosing of cyclophosphamide for establishing long-lasting skin allograft survival, stable mixed chimerism, and intrathymic clonal deletion in mice primed with allogeneic spleen cells.

BACKGROUND: Injection of allo-spleen cells (SC) followed by a single dose of cyclophosphamide (CP) can induce tolerance of tumor and/or skin allografts in mice. To minimize the damage caused by CP, fractionation of CP that can establish long-lasting skin graft survival, stable mixed chimerism, and intrathymic clonal deletion in the host was investigated in the present study. METHODS: Allo-SC (10(8)) were given intravenously on day 0. CP at 200 mg/kg was given intraperitoneally on day 2 in a single dose (CP 200x1 group). CP at 100, 66, 50, 40, and 33 mg/kg was given daily from day 1 through days 2, 3, 4, 5 and 6, respectively, in the fractionated doses (CP 100x2, 66x3, 50x4, 40x5, and 33x6 groups; total dose=200 mg/kg). Allografting was performed on day 14. RESULTS: In a fully allogeneic combination of C57BL/6 (H2b)-->AKR (H2k, Mls-1a), an EL-4 tumor (H2b) was specifically accepted to kill the AKR mice in all of the SC+CP 200x1, 100x2, 66x3, 50x4, 40x5, and 33x6 groups (n=6), but C57BL/6 skin graft survival was not prolonged in any of the tumor-tolerant groups. In an H2-identical combination of AKR-->C3H (H2k, Mls-1b), AKR skin graft survival was prolonged remarkably (80-90 days) in the SC+CP 200x1, 100x2, and 66x3 groups (n=5-11), but was prolonged moderately (20-60 days) in the SC+CP 50x4 and 40x5 groups. In both of the SC+CP 200x1 and 66x3 groups in the AKR-->C3H combination, mixed chimerism was maintained for as long as 100 days after tolerance induction in both the spleen and thymus, associated with intrathymic clonal deletion of Vbeta6+ T cells. The decreases in leukocyte count, hemoglobin level, spleen weight, SC count, and body weight were significantly smaller in the SC+CP 66x3 group than in the SC+CP 200x1 group. CONCLUSIONS: Fractionated CP is effective in ameliorating the compromised state induced by a single dose of CP. To induce a long-lasting skin allograft survival associated with stable mixed chimerism and intrathymic clonal deletion in an H2-identical combination, 200 mg/kg of CP can be divided into three or fewer fractions.

Improved technique of heterotopic cervical heart transplantation in mice.

BACKGROUND: The method for mouse vascularized heart transplantation have been described using suture and cuff techniques. Technical problems have limited its widespread use. Here, we describe our method of modified cervical heterotopic transplantation with the cuff technique. METHODS: By using a smaller Teflon cuff (external diameter 0.6 mm, internal diameter 0.4 mm) and superfine-tip forceps, it became possible to directly pull the edge of the carotid artery and evert the proximal end of the artery over the cuff. Similarly, the external jugular vein could be easily everted over a 22-gauge cuff with this direct pulling method. RESULTS: By these modifications, the operation time was reduced. It usually takes 20 min for the donor harvest, 15 min for preparation of the cervical vessels, and 15 min for anastomosis. All procedures from the donor harvest through skin closure of the recipient mice can be completed within 1 hr, and ischemic time is within 25-40 min. CONCLUSIONS: This method can be used to investigate cyclophosphamide-induced tolerance and mechanisms of reperfusion injury.