[Epidemiological and molecular characteristics of human brucellosis in Qinghai province, 2005-2019].

Objective: To analyze the epidemiological and molecular characteristics of human brucellosis in Qinghai province from 2005 to 2019 and provide basic data for brucellosis prevention and control. Method: The data about human brucellosis in Qinghai from 2005 to 2019 were collected from the information system of China CDC to describe the spatial, population and time distributions of human brucellosis cases in Qinghai. The isolated strains were identified and typed with traditional methods, BCSP31-PCR, AMOS-PCR and multi-locus variablenumber tandem repeat (MLVA-16). Results: A total of 577 human brucellosis cases were reported in Qinghai from 2005 to 2019, the average prevalence rate was 0.07 per 100 000 person, there were statistic differences among different years. The disease occurred all the year around, but mainly during March-October. The 577 cases were distributed in 31 counties (cities/districts) from 6 autonomous prefectures (cities). The prevalence rats of five counties were high, i.e. Menyuan Hui autonomous county (22.88%, 132/577), Tianjun county (10.57%, 61/577)、Xining city (10.57%, 61/577), Henan Mongol Autonomous County (10.51%, 58/577) and Haiyan county (9.53%, 55/577). Age of the cases ranged from 8 years to 82 years, and the male to female ratio of the cases was 1.8∶1 (374/203). The prevalence rate in herdsman (47.83%, 276/577) was highest among different occupational populations. Ten isolates were all Brucella melitensis strains, belonging to biovar 3, and clustering analysis indicated that the 10 strains had 5 genotypes, in which 2 were distinct, the remaining 3 were same. MLVA-16 analysis indicated that the 10 strains had close relationship with 26 B. melitensis strains isolated in Qinghai previously. Conclusions: The prevalence of brucellosis increased in Qinghai in recent years, we should strengthen the population based brucellosis surveillance and reporting. MLVA-16 indicated the gene diversity of the Brucella strains, suggesting that MLVA-16 can be used for genetic diversity analysis and molecular epidemiology survey to improve brucellosis surveillance.

[Epidemiological characteristics of Brucella species isolated from different regions of the world using the MLVA genotyping].

Objective: To study the molecular-epidemiological characteristics of Brucella species isolated from different countries, using the multiple locus tandem-repeat (MLVA) analysis. Methods: Eleven variable-number tandem-repeat (VNTR) loci were selected. VNTR strains of Brucella isolated from 48 different countries in 1953-2013, were analyzed by using the BioNumerics software. Unweighted Paired Arithmetic Average method was used to cluster and draw phylogenetic tree as well as the minimum spannin. Results: The evolutionary relationship of Brucella phylogenetic tree was consistent with the classical biological typing method. However, the Brucella suis biovar 5 strains were different from the other Brucella suis biovars 1, 2, 3 and 4. Brucella ceti strains were divided into two parts and different from each other. Worldwide epidemics of brucellosis were emerged from 2005 to 2008 under the MLVA11 Orsay analysis. China has been a brucellosis-prone regions, with Brucella melitensis as the main epidemic Brucella species, followed by Brucella abortus. Brucella suis was mainly identified in the southern provinces, but Brucella canis was mainly found in dogs. No human cases were found. Conclusion: Molecular-epidemiological characteristics of the Brucella strains were related to factors as time, region and hosts of isolation, which are important to setting up prevention and control programs on brucellosis.

Identification of critical microRNAs in gastrointestinal stromal tumor patients treated with Imatinib.

Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal neoplasms of the gastrointestinal tract. Imatinib mesylate was considered to be a breakthrough drug in clinical treatment of GIST, but GIST patients showed resistance against it. We aimed to identify critical microRNAs (miRNAs) related to imatinib resistance in imatinib-treated GIST patients. Microarray datasets under the accession number of GSE63159 and GSE45901 were downloaded from the Gene Expression Omnibus (GEO) database. The differentially expressed miRNAs (DEMs) that are related to imatinib resistance were identified. GO function and KEGG pathway enrichment analyses were performed, and lncRNA-miRNA-target gene regulatory networks were constructed. Finally, the critical miRNAs and their target genes that are related to imatinib resistance or sensitivity were identified. In total, 20 DEMs in the GSE63159 dataset (7 significantly up-regulated and 13 down-regulated) and 23 DEMs in the GSE45901 dataset (8 up-regulated and 15 down-regulated) were identified. In lncRNA-miRNA-target gene regulatory networks, five critical miRNAs and 109 target genes were identified. GO function and KEGG pathway enrichment analysis showed that the target genes of DEMs were mainly involved in several signaling pathways, such as focal adhesion and the GnRH signaling pathway. Among the five miRNAs, the overexpression of hsa-miR-28-5p and hsa-miR-125a-5p had significant correlation to imatinib resistance or imatinib sensitivity in GIST patients. Hsa-miR-28-5p and hsa-miR-125a-5p may be involved in the development and progression of GIST, and they may be able to serve as prognostic markers for imatinib-response in GIST patients.

ARID1A gene knockdown promotes neuroblastoma migration and invasion.

Neuroblastoma is the most common extracranial solid tumor in childhood which often acquires drug resistance and becomes aggressive phenotypes. The high-risk patients suffer from high mortality due to the limitation of the treatment strategies. ARID1A (AT-rich interactive domain-containing protein 1A), a subunit of SWI/SNF complexes, is considered as a tumor suppressor in many cancers. The aim of the present study was to investigate the effect of ARID1A on migration and invasion in neuroblastoma cells. The shRNA targeting ARID1A was designed and delivered into SK-N-SH cells to knock down ARID1A expression. Knockdown of ARID1A by shRNA significantly increased the viability and invasion ability, and caused G1 arrest inhibition and DNA synthesis increase in SK-N-SH cells. Moreover, Knockdown of ARID1A increased the activity and expression of matrix metalloproteinase (MMP)-2 and -9 in SK-N-SH cells. Furthermore, ARID1A knockdown caused diminished expression of E-cadherin, enhanced expression of N-cadherin and ß-catenin nuclear translocation in SK-N-SH cells. These results suggest that loss of ARID1A may associate with the promotion of invasion and metastasis of neuroblastoma. Our findings indicate ARID1A is a tumor suppressor in neuroblastoma.

Hyperthermia sensitization and proton beam triggered liposomal drug release for targeted tumor therapy.

PURPOSE: The objectives of this study were to: 1) determine if mild hyperthermia (40-42°C) can sensitize tumor cells for more effective proton beam radiotherapy (PBRT); 2) characterize the survival fraction of cells exposed to PBRT; and 3) characterize release of the drug doxorubicin (Dox) from low temperature sensitive liposomes (LTSLs) without exposure to mild hyperthermia in combination with PBRT. METHODS: Dox was actively loaded in LTSLs. A549 monolayer cells were incubated with 100-200 nM of Dox-LTSL (±mild hyperthermia). Cell irradiation (0-6 Gy) was performed by placing the cell culture plates inside a solid water phantom and using a clinical proton treatment beam with energy of 150 MeV. End points were survival fraction, radiation-mediated Dox release, and reactive oxygen species (ROS) production. RESULTS: Hyperthermia effectively sensitized cells for PBRT and lowered the cell survival fraction (SF) by an average of 9.5%. The combination of 100 nM Dox-LTSL and PBRT (1-6 Gy) achieved additive to synergistic response at various dose combinations. At higher radiation doses (>3 Gy), the SF in the Dox and Dox-LTSL groups was similar (~20%), even in the absence of hyperthermia. In addition, 30% of the Dox was released from LTSLs and a 1.3-1.6 fold increase in ROS level occurred compared to LTSL alone therapy. CONCLUSIONS: The combination of LTSLs and PBRT achieves additive to synergistic effect at various dose combinations in vitro. Concurrent PBRT and Dox-LTSL treatment significantly improved the cytotoxic outcomes of the treatment compared to PBRT and Dox chemotherapy without LTSLs. We hypothesize that PBRT may induce drug release from LTSL in the absence of hyperthermia.

Transrectal ultrasound-integrated spectral optical tomography of hypoxic progression of a regressing tumor in a canine prostate.

The objective of this study was to evaluate if transrectal optical tomography implemented at three wavelength bands for spectral detection could monitor changes of the hemoglobin oxygen saturation (StO2) in addition to those of the total hemoglobin concentration ([HbT]) in lesions of a canine prostate, including an induced tumor modeling canine prostate cancer. Near-infrared (NIR) optical tomography was integrated with ultrasound (US) for transrectal imaging. Multi-spectral detection at 705_nm, 785_nm and 808_nm rendered measurements of [HbT] and StO2. Canine transmissible venereal tumor (TVT) cells were injected into the right lobe of a dog's prostate gland, which had a pre-existing cyst in the left lobe. Longitudinal assessments of the prostate were performed weekly over a 63-day duration by NIR imaging concurrent with grey-scale and Doppler US. Ultrasonography revealed a bi-lobular tumor-mass regressing from day-49 to day-63. At day-49 this tumor-mass developed a hypoxic core that became larger and more intense by day-56 and expanded further by day-63. The tumor-mass presented a strong hyper-[HbT] feature on day-56 that was inconsistent with US-visualized blood flow. Histology confirmed two necrotic TVT foci within this tumor-mass. The cyst appeared to have a large anoxic-like interior that was greater in size than its ultrasonographically delineated lesion, and a weak lesional elevation of [HbT]. On day-56, the cyst presented a strong hyper-[HbT] feature consistent with US-resolved blood flow. Histology revealed acute and chronic hemorrhage in the periphery of the cyst. The NIR imaging features of two other TVT nodules and a metastatic lymph node were evaluated retrospectively. Transrectal US-integrated spectral optical tomography seems to enable longitudinal monitoring of intra-lesional oxygenation dynamics in addition to the hemoglobin content of lesions in the canine prostate.

Trans-rectal ultrasound-coupled spectral optical tomography of total hemoglobin concentration enhances assessment of the laterality and progression of a transmissible venereal tumor in canine prostate.

OBJECTIVES: To evaluate whether trans-rectal spectral optical tomography of total hemoglobin concentration (HbT) can image longitudinal and lateral developments of a canine transmissible venereal tumor (TVT) in a canine prostate. METHODS: A near-infrared (NIR) applicator was integrated with a trans-rectal ultrasound (TRUS) transducer to perform ultrasound (US)-coupled optical tomography of the canine prostate. Spectral detection at 785 and 830 nm enabled quantitation of HbT. Canine TVT cells were injected into the right lobe of a dog's prostate gland. Longitudinal imaging assessment of the post-injection prostate was performed by coupled US/NIR imaging over a 45-day duration. RESULTS: By day 7, NIR indicated TVT infiltration in the noninjected left prostatic lobe with the gray-scale US indistinct. By day 31, both NIR and gray-scale US revealed more widespread TVT involvement in the left than in the right lobe, as well as an extensive TVT mass in the caudal aspect of the gland, of which the peak HbT increased 3-fold and the mass volume grew exponentially over the 45-day duration. Increased blood supply to the mass was also observed on Doppler US. CONCLUSIONS: TRUS-coupled spectral optical tomography enhances assessment of the laterality and progression of prostate tumor compared with using gray-scale and Doppler TRUS.

Design of near-infrared imaging probe with the assistance of ultrasound localization.

A total of 364 optical source-detector pairs were deployed uniformly over a 9 cm x 9 cm probe area initially, and then the total pairs were reduced gradually to 60 in experimental and simulation studies. For each source-detector configuration, three-dimensional (3-D) images of a 1-cm-diameter absorber of different contrasts were reconstructed from the measurements made with a frequency-domain system. The results have shown that more than 160 source-detector pairs are needed to reconstruct the absorption coefficient to within 60% of the true value and appropriate spatial and contrast resolution. However, the error in target depth estimated from 3-D images was more than 1 cm in all source-detector configurations. With the a priori target depth information provided by ultrasound, the accuracy of the reconstructed absorption coefficient was improved by 15% and 30% on average, and the beam width was improved by 24% and 41% on average for high- and low-contrast cases, respectively. The speed of reconstruction was improved by ten times on average.

Cancellation of coherent artifacts in optical coherence tomography imaging.

Coherent artifacts in optical coherence tomography (OCT) images can severely degrade image quality by introducing false targets if no targets are present at the artifact locations. Coherent artifacts can also add constructively or destructively to the targets that are present at the artifact locations. This constructive or destructive interference will result in cancellation of the true targets or in display of incorrect echo amplitudes of the targets. We introduce the use of a nonlinear deconvolution algorithm, CLEAN, to cancel coherent artifacts in OCT images of extracted human teeth. The results show that CLEAN can reduce the coherent artifacts to the noise background, sharpen the air-enamel and enamel-dentin interfaces, and improve the image contrast.

Simultaneous near-infrared diffusive light and ultrasound imaging.

We have constructed a near-real-time combined imager suitable for simultaneous ultrasound and near-infrared diffusive light imaging and coregistration. The imager consists of a combined hand-held probe and the associated electronics for data acquisition. A two-dimensional ultrasound array is deployed at the center of the combined probe, and 12 dual-wavelength laser source fibers (780 and 830 nm) and 8 optical detector fibers are deployed at the periphery. We have experimentally evaluated the effects of missing optical sources in the middle of the combined probe on the accuracy of the reconstructed optical absorption coefficient and assessed the improvements of a reconstructed absorption coefficient with the guidance of the coregistered ultrasound. The results have shown that, when the central ultrasound array area is in the neighborhood of 2 cm x 2 cm, which corresponds to the size of most commercial ultrasound transducers, the optical imaging is not affected. The results have also shown that the iterative inversion algorithm converges quickly with the guidance of a priori three-dimensional target distribution, and only one iteration is needed to reconstruct an accurate optical absorption coefficient.

Modified cold storage of rat livers with self-made HYD solution.

OBJECTIVE: To investigate the cold preservation effect on rat livers of a modified storage method with self-made HYD solution. METHODS: The vascular bed of rat livers was expanded with an additional 20 to 40 ml self-made HYD solution/100 g liver. After resection of the liver, the extra HYD solution (expressed as % liver weight) was entrapped via portal infusion by tying off the supra- and infra-hepatic inferior vena cava. Forty rats were randomly divided into four groups including control group with conventional storage method, and 20%, 30% and 40% groups according to the amount of extra HYD solution. We compared the preservation effect of the modified storage method with that of the conventional storage method using an isolated perfused rat liver model. RESULTS: Bile production and all the indices of hepatic microcirculation including portal perfusion pressure, endothelin-1 in the effluent, trypan blue distribution time and histology were significantly superior in the modified method groups compared to those in the control group (P < 0.05). The contents of dihydroxybenzoic acid (DHBA) in the modified method groups were significantly lower than those in the control group (P < 0.05). Liver enzymes activities in the 30% group were markedly lower than those in the control group (P < 0.05). The preservation effect on rat liver in the 30% group was the best among the modified method groups. CONCLUSION: The modified cold storage method is effective and may have potential for clinical application in liver preservation.

Clinical application of serial operations with preserving spleen.

AIM: To evaluate the clinical application of serial operations with preservation of spleen. METHODS: Serial operations with preserving spleen were performed on 211 cases in our hospital from 1980 to 2000. The patient's age ranged from 13 to 56 years, averaging 38 years. Diseases included splenic injury in 171 cases, portal hypertension in 9 cases, splenic cyst in 10 cases, and the lesion of pancreatic body and tail in 21 cases. RESULTS: All the cases were cured, and 129 patients were followed up from 3 months to 3 years with the leukocyte phagocytosis test, detection of immunoglubin, CT,(99m)Tc scanning and ultrasonography. The results were satisfactory. CONCLUSION: The operations with preserving spleen were safe, feasible, and worth of clinical application.

New method for preparation of coumarins and quinolinones via Pd-catalyzed intramolecular hydroarylation of C-C triple bonds.

A new and general method has been developed for preparation of coumarins and quinolinones by intramolecular hydroarylation of alkynes. Various aryl alkynoates and alkynanilides undergo fast intramolecular reaction at room temperature in the presence of a catalytic amount of Pd(OAc)(2) in a mixed solvent containing trifluoroacetic acid (TFA), affording coumarins and quinolinones in moderate to excellent yields with more than 1000 turnover numbers (TON) to Pd. The methodology proved to tolerate a number of functional groups such as Br and CHO. On the basis of isotope experiments, a possible mechanism involving ethynyl chelation-assisted electrophilic metalation of aromatic C-H bonds by in-situ generated cationic Pd(II) species has been discussed. Also the involvement of vinylcationic species has been suggested.

Efficient activation of aromatic C-H bonds for addition to C-C multiple bonds

Efficient electrophilic metalation of aromatic C-H bonds leading to new C-C bond formation through regio- and stereoselective addition to alkynes and alkenes has been realized by a catalytic amount (0.02 to 5 mole percent) of palladium(II) or platinum(II) compounds in a mixed solvent containing trifluoroacetic acid at room temperature. Various arenes undergo unexpected selective trans hydroarylation to terminal or internal C&cjs0812;C bonds inter- and intramolecularly with high efficiency (up to a turnover number of 4500 for palladium), especially for electron-rich arenes, giving thermodynamically unfavorable cis-alkenes, and the oxygen- and nitrogen-containing heterocycles. The simplicity, generality, and efficiency of this process should be very attractive to the possible industrial application for the functionalization of arenes.

Study on modified cold storage method of rat livers with self-made hyd solution.

OBJECTIVE: To investigate the cold preservation effect of rat livers by modified storage method with self-made HYD solution. METHODS: The modified method was that the vascular bed of rat livers was expanded with an additional 20 to 40ml self-made HYD solution/100g liver. After removing the liver, the extra HYD solution expressed as % liver weight was entrapped via portal infusion by tying off the supra-and infra hepatic inferior vena cava. According to the amount of extra HYD solution, 40 rats were randomly divided into four groups including: control group with conventional storage method, 20% group, 30% group and 40% group. The preservation effect of modified storage method with that of conventional storage method by using isolated perfused rat liver model was compared. RESULTS: Bile production and all the indices of hepatic microcirculation including portal perfusion pressure, endothelin-1 in the effluent, trypan blue distribution time and histology in modified method groups were significantly superior to those in control group (P < 0.05). The liver enzymes in 30% group were markedly lower than those in control group (P < 0.05). The preservation effect of rat liver in 30% group was the best among the modified method groups. CONCLUSION: The modified cold stage method is effective and may have potential for clinical application for liver preservation.

Cytoplasmic delayed neuronal death in the myenteric plexus of the rat small intestine after ischemia.

The present study demonstrates light and electron microscopic changes in neurons in the myenteric plexus of the rat ileum following four-hour ischemia. Macroscopically, an intestinal constriction occurred at the damaged portion at three weeks after ischemia; the segment oral to the constriction markedly swelled at four weeks. In light microscopy, at three weeks after ischemia, the myenteric neurons appeared spongy or foamy, containing many vacuoles in their somatic cytoplasm. At four weeks, the neuronal cytoplasm and nerve fiber bundles had disintegrated to form vacant spaces in the myenteric plexus. The neuronal nucleus of the damaged plexus did not show positive nick-end labeling. In electron microscopy, neuronal cytoplasm revealed degenerative signs already at one week after ischemia: a distended endoplasmic reticulum and swollen mitochondria with fragmentary cristae. The nerve fibers also showed destruction of the mitochondria, and degenerative changes in the postsynaptic sites appeared earlier than the presynaptic terminals. The results suggest that intestinal ischemia causes delayed neuronal death, which differs from the apoptotic process previously demonstrated in the ischemia-damaged brain.

Typology of abdominal arteries, with special reference to inferior phrenic arteries and their esophageal branches.

Origins and distribution of the human inferior phrenic arteries were studied by dissecting 68 Japanese adult cadavers. The inferior phrenic arteries were usually observed as paired (left and right) vessels. Their origins were summarized as follows: a) the aorta itself (85/138 cases, 61.6%), b) the ventro-visceral arteries (celiaco-mesenteric system of the aorta) including the celiac trunk (39/138 cases, 28.2%) and the left gastric artery (4/138 cases, 2.9%), and c) the latero-visceral arteries (adreno-renal system of the aorta) including the middle adrenal artery (4/138 cases, 2.9%) and the renal artery (6/138 cases, 4.3%). The left and right arteries occasionally originated in common trunk from the aorta, celiaco-mesenteric system or adreno-renal system (22/138 cases, 15.9%). A typological diagram explaining these variations is given. The inferior phrenic arteries, especially the left ones, sometimes issued visceral or esophageal branches. This fact indicates that the inferior phrenic arteries are homologous with the celiac trunk and mesenteric arteries. It is further discussed that the celiac trunk and mesenteric arteries are originally paired vessels, through introduction of our previous typological diagram of the abdominal arteries.

Perineuronal sulfated proteoglycans and cell surface glycoproteins in adult and newborn mouse brains, with special reference to their postnatal developments.

Sections of the retrosplenial cortex from adult and newborn mouse brains were observed with a light microscope. The retrosplenial cortex of the adult animals contained many neurons (10% of the total), including some dark neurons, with perineuronal sulfated proteoglycans detectable with cationic iron colloid and aldehyde fuchsin. The retrosplenial cortex of the adult animals also contained many neurons (10% of the total) with cell surface glycoproteins reactive to lectin Vicia villosa, soybean or Wisteria floribunda agglutinin. Double staining showed that the majority (75%) of the neurons labeled with lectins were stained with cationic iron colloid, and that some (25%) of them were not stained with this colloid. Double staining also showed that some (25%) of the neurons stained with cationic iron colloid were not labeled with lectins. These findings indicate that the perineuronal sulfated proteoglycans are, at least partly, independent from the cell surface glycoproteins. Observations of the sections from the newborn animals revealed that the perineuronal sulfated proteoglycans were produced by the associated satellite astrocytes 3-4 weeks after birth, and that the cell surface glycoproteins were produced by the associated nerve cells at earlier stages, or 2-3 weeks after birth. Dark neurons began to appear 3-4 weeks after birth. These dark neurons or their Golgi complexes were also reactive to lectins, suggesting the production of cell surface glycoproteins.

Perineuronal sulfated proteoglycans in the human brain are identical to Golgi's reticular coating.

Many neurons in the human somatosensory cortex (Area 7 of Brodmann) possess an intensely negative-charged surface coat consisting of perineuronal sulfated proteoglycans which were stained with cationic iron colloid. This surface coat was stained doubly with cationic iron colloid and Golgi's silver nitrate. The result indicates that the perineuronal sulfated proteoglycans are identical with Golgi's reticular coating, whose existence and nature have previously been controversial. The result also suggests that Golgi's silver nitrate stains the core proteins of proteoglycans.

Bicuspid aortic valve in an adult human cadaver: a case report.

We encountered a "bicuspid" aortic valve in an 83-year-old man. This valve consisted of two semilunar leaflets, anterior and posterior, though they were extensively calcified. The right and left coronary arteries independently arose from the anterior aortic sinus. The infundibular ligament ran from the pulmonary trunk at the commissure of the right and left semilunar cusps to the aorta at the mid-point of the anterior sinus. These findings strongly suggests the present bicuspid aortic valve may be formed by fusion of usual right and left (coronary) cusps. No other congenital anomalies were noted in this heart.