Prenatal diagnosis of fetal conotruncal defects by using 2D ultrasound and HD live flow combined with spatiotemporal image correlation.

INTRODUCTION: To explore the diagnostic value of spatiotemporal image correlation (STIC) for different types of fetal conotruncal defects (CTDs). METHODS: The clinical data and STIC images of 174 fetuses with CTDs diagnosed via prenatal ultrasound were analyzed retrospectively. RESULTS: Among the 174 cases of CTDs, 58 were tetralogy of Fallot (TOF); 30, transposition of great arteries (TGA) (D-TGA, 23 cases; cc-TGA, 7 cases); 26, double outlet of the right ventricle (DORV); 32, persistent arterial trunk (PTA) (type A1, 15 cases; type A2, 11 cases; type A3, 5 cases; type A4, 1 case); and 28, pulmonary atresia (PA) (ventricular septal defect, 24 cases; ventricular septal integrity, 4 cases). Among the cases, 156 were complicated with complex congenital intracardiac and extracardiac malformations. The abnormal display rate of the four-chamber view of two-dimensional echocardiography was low. The display rate of the permanent arterial trunk was the highest (90.6%) in STIC imaging. CONCLUSIONS: STIC imaging can be used in the diagnosis of different types of CTDs, especially in persistent arterial trunks, and thus has great value for the clinical treatment and prognosis of these defects.

High selectivity of photocatalytic reduction of CO2 to CO based on terpyridine ligand supported CuI metal organic framework.

In this work, a 4'-(4-cynaophenyl)-4,2':6',4-terpyridine supported CuI MOFs photocatalyst (Cu I MOF) was applied to the photocatalytic CO2 reduction for the first time. The micro-structural and physicochemical properties of the Cu I MOF were systematically studied by the powder X-ray diffraction (PXRD), Single crystal X-ray diffraction (SCXRD), scanning electron microscope (SEM), X-ray photoelectron spectroscopy (XPS), Fourier transform infrared (FT-IR), UV-Vis diffuse spectroscopy (UV-vis DRS), ns-level photoluminescence spectra (ns-level PL), Ultraviolet photoelectron spectroscopy (UPS), and N2 adsorption-desorption test (BET-BJH). Moreover, the in situ diffuse reflectance infrared fourier transform spectroscopy (in situ DRIFTS) was applied to investigate the adsorption and reaction intermediates of photocatalytic CO2 reduction. As a result, Cu I MOF exhibited good performance and outstanding selectivity toward photocatalytic reduction of CO2 to CO under full-spectrum and visible light illumination. Notably, 100% selective photocatalytic conversion of CO2 to CO was achieved. Thus, the study presents the high selectivity and CO2 reduction efficiency of Cu I MOF as a potential family of photocatalysts.

An actively compensated 8 nT-level magnetic shielding system for 10-m atom interferometer.

We design and develop a high-performance magnetic shielding system for a long baseline fountain-type atom interferometer. The shielding system is achieved by a combination of passive shielding using permalloy and active compensation with coils. An 11.4 m-long three-layer cylindrical shield is completed by the process of welding, local annealing, and entire annealing. The active compensations compress the residual magnetic field to 8.0 nT max-to-min and the corresponding gradient below 30 nT/m over 10 m along the axial direction in which external compensation, internal compensation, and constant magnetic field (C-field) compensation reduce the inhomogeneities to 25.0, 12.6, and 1.7 nT (standard deviation) sequentially. We estimate that this system could reduce the systematic error of the quadratic Zeeman shift to the 10-13 level for the weak equivalence principle test with a simultaneous 85Rb-87Rb dual-species atom interferometer.

Finite element analysis of wedge and biconcave deformity in four different height restoration after augmentation of osteoporotic vertebral compression fractures.

PURPOSE: Biomechanical comparison of wedge and biconcave deformity of different height restoration after augmentation of osteoporotic vertebral compression fractures was analyzed by three-dimensional finite element analysis (FEA). METHODS: Three-dimensional finite element model (FEM) of T11-L2 segment was constructed from CT scan of elderly osteoporosis patient. The von Mises stresses of vertebrae, intervertebral disc, facet joints, displacement, and range of motion (ROM) of wedge and biconcave deformity were compared at four different heights (Genant 0-3 grade) after T12 vertebral augmentation. RESULTS: In wedge deformity, the stress of T12 decreased as the vertebral height in neutral position, flexion, extension, and left axial rotation, whereas increased sharply in bending at Genant 0; L1 and L2 decreased in all positions excluding flexion of L2, and T11 increased in neutral position, flexion, extension, and right axial rotation at Genant 0. No significant changes in biconcave deformity. The stress of T11-T12, T12-L1, and L1-L2 intervertebral disc gradually increased or decreased under other positions in wedge fracture, whereas L1-L2 no significant change in biconcave fracture. The utmost overall facet joint stress is at Genant 3, whereas there is no significant change under the same position in biconcave fracture. The displacement and ROM of the wedge fracture had ups and downs, while a decline in all positions excluding extension in biconcave fracture. CONCLUSIONS: The vertebral restoration height after augmentation to Genant 0 affects the von Mises stress, displacement, and ROM in wedge deformity, which may increase the risk of fracture, whereas restored or not in biconcave deformity.

Performance of CNTs/GQD-Based Flexible Strain Sensors.

Nanomaterial-based flexible strain sensors have developed rapidly in recent years. Here, we propose a flexible strain sensor based on polydimethylsiloxane with carbon nanotubes (CNTs) and graphene quantum dots (GQDs). Different weight ratios of CNTs and GQDs were used as the sensitive units of the strain sensors. After analyzing the results of current-voltage curves and the strain effects of the sensors, we concluded that the introduction of GQDs played an important role in improving the sensitivity of the sensors. The gauge factor of the as-prepared strain sensors ranges from 0 to 841.42.

[Treatment of Vascular Cognitive Impairment by "Huayu Tongluo" Moxibustion].

OBJECTIVE: To observe the therapeutic effect of "Huayu Tongluo"(blood stasis-removing and meridian-collateral-dredging) moxibustion for vascular cognitive impairment(VCI) patients and changes of insulin like growth factor -1(IGF-1) levels in serum after the treatment. METHODS: Sixty patients with VCI were randomly divided into medication (control) and moxibustion groups (n＝30 in each group). Cotton cloth-separated moxibustion was applied to Baihui (GV 20) and Shenting (GV 24), and conventional moxibustion applied to Dazhui (GV 14) and Yongquan (KI 1) for 30 min, once daily, 6 times a week and for 30 days. Patients of the control group were treated by oral administration of Donepezil hydrochloride at the dose of 5 mg/night for 30 days. The core symptoms of traditional Chinese medicine (TCM), mini-mental state examination(MMSE), activity of daily living(ADL) and Montreal cognitive assessment(MoCA) scales were used to assess the therapeutic effect after the treatment. The content of serum IGF-1 was determined by ELISA. RESULTS: Of the two 30 cases in the control and moxibustion groups, 9 and 17 experienced marked improvement, 13 and were effective, 8 and 3 ineffective, with the effective rates being 73.33% and 90.00%, respectively. The effective rate in the moxibustion group was obviously higher than that in the control group (P<0.05). After the treatment, the TCM symptom scores were significantly decreased, and the MMSE, ADL and MoCA scores considerably increased in both groups compared with those of their own individual pre-treatment (P<0.01). The TCM symptom score of the moxibustion group was significantly lower, and the MMSE and ADL scores were obviously higher than those of the control group (P<0.01). The serum IGF-1 content in both groups was significantly increased after the treatment relevant to that of their own individual pre-treatment (P<0.01), and was obviously higher in the moxibustion group than in the control group (P<0.01). No significant difference was found between the two groups in the MoCA score after the treatment (P>0.05)．. CONCLUSION: "Huayu Tongluo" moxibustion has a positive effect for patients with VCI, which may be associated with its effect in up-regulating serum IGF-1 level.

[Effect of water-nitrogen coupling on photosynthesis and ultrastructure of cucumber leaves under CO2 enrichment]. / CO2åŠ å¯Œä¸‹æ°´æ°®è€¦åˆå¯¹é»„ç“œå¶ç‰‡å ‰åˆä½œç”¨å’Œè¶ å¾®ç»“æž„çš„å½±å“.

Using split plot and then-split plot design, effects of water-nitrogen coupling on photosynthesis and ultrastructure of cucumber (Cucumis sativus) (Jinyou No.35) under CO2 enrichment were investigated. The main plot had two CO2 concentrations: ambient CO2 concentration (400 µmol·mol-1, A) and doubled CO2 concentration (800±20 µmol·mol-1, E). The split plot had two treatments: no drought stress (95% of field capacity, W) and drought stress (75% of field capacity, D). The then-split plot contained low nitrogen treatment (450 kg·hm-2, N1) and high nitrogen treatment (900 kg·hm-2, N2). The results showed that under the condition of drought and high nitrogen, increasing CO2 enhanced the cucumber plant height, and no matter what kinds of water treatment, CO2 enrichment increased the leaf area significantly under high nitrogen. Under the condition of normal irrigation, the photosynthetic rate, stomatal conductance and transpiration rate of high nitrogen treatment were higher than low nitrogen treatment, while it was under the drought condition. Elevated CO2 enhanced the water use efficiency of cucumber leaf which increased with increasing nitrogen application rate. Under drought stress, cucumber adaxial surface porosity density was increased, and the CO2 enrichment and high nitrogen significantly reduced the stomatal density. Increasing nitrogen application improved the number of chloroplast, and reduced that of starch grains. Drought stress decreased the number of chloroplast, but tended to promote the number of starch grains. Drought stress increased the chloroplast length and width, and the size of the starch grains, while high nitrogen reduced the length and width of the chloroplast and starch grains. CO2 enrichment and high nitrogen increased grana thickness and layers (except ADN2), and the slice layer of EDN2 was significantly higher than that of ADN2. In conclusion, CO2 enrichment and suitable water and nitrogen could promote the development of chloroplast thylakoid membrane system, significantly increase the thickness of grana and the number of grana lamella, and effectively improve the chloroplast structure of cucumber, which would benefit the photosynthesis of cucumber plants and ability to utilize CO2 and water and nitrogen.

Lipophilic but not hydrophilic statin functionally inhibit volume-activated chloride channels by inhibiting NADPH oxidase in monocytes.

Volume-activated Cl- channels (VACCs) can be activated by hypotonic solutions and have been identified in many cell types. Here, we investigated the effects of different statins on VACCs in monocytes. Whole-cell patch clamp recordings demonstrated that a hypotonic solution induced 5-nitro-2- (3-phenylpropylamino) benzoic acid (NPPB)- and 4,4'-diisothiocyanatostilbene-2, 2'-disulfonic acid (DIDS)-sensitive VACC currents in human peripheral monocytes and RAW 264.7 cells. The VACC currents were inhibited by the lipophilic statin (simvastatin) but not by the hydrophilic simvastatin acid and pravastatin. A low-molecular-weight superoxide anion scavenger (tiron, 1 mM) and inhibitor of NADPH oxidase (DPI 10 µM) was able to abolish the VACC currents. A hypotonic solution increased the reactive oxygen species (ROS) detected by the fluorescence of dichlorodihydrofluorescein (DCF), which was abolished by tiron and DPI. NPPB, DIDS, and simvastatin but not pravastatin decreased the fluorescence of DCF. Simvastatin could not further decrease VACC currents when pretreated with tiron or DPI, whereas exogenous H2O2 (100 µM), increased the VACC currents and overcame the blockade of VACC currents by simvastatin. Functionally, hypotonic solution increased the TNF-α mRNA expression, which could be decreased by tiron, DPI, NPPB, DIDS and simvastatin but not pravastatin. However, simvastatin could not decrease the TNF-α expression further when pretreatment with tiron, DPI, NPPB or DIDS. We conclude that lipophilic (simvastatin) rather than hydrophilic statin inhibit VACCs and decrease hyposmolality induced inflammation in monocytes by inhibiting NADPH oxidase.

[Study on Strain Detection with Si Based on Bicyclic Cascade Optical Microring Resonator].

Optical micro-ring resonator prepared on Silicon-On-Insulator (SOI) has high sensitivity, small size and low mode volume. Its high sensitivity has been widely applied to the optical information transmission and inertial navigation devices field, while it is rarely applied in the testing of Mechanics. This paper presents a cantilever stress/strain gauge with an optical microring resonator. It is proposed the using of radius change of ring waveguide for the sensing element. When external stress is put on the structure, the radius of the SOI ring waveguide will be subjected to variation, which causes the optical resonant parameters to change. This ultimately leads to a red-shift of resonant spectrum, and shows the excellent characteristics of the structure's stress/strain sensitivity. Designed a bicyclic cascade embedded optical micro-cavity structure, which was prepared by employing MEMS lithography and ICP etching process. The characteristic of stress/strain sensitivity was calculated theoretically. Two values of 0.185 pm x kPa(-1) and 18.04 pm x microstrain(-1) were obtained experimentally, which also was verified by theoretical simulations. Comparing with the single-loop micro-cavity structure, its measuring range and stress sensitivity increased by nearly 50.3%, 10.6%, respectively. This paper provides a new method to develop micro-opto-electromechanical system (MOEMS) sensors.

Hepatocellular carcinoma metastasis to the gingival soft tissues: A case report and review of the literature.

Metastases to the gingival soft tissues are rare in hepatocellular carcinoma (HCC). To the best of our knowledge, only 13 cases have been reported in English literature to date. The present study described the case of a 43-year-old Chinese man who was admitted to Tangdu Hospital (Xi'an, China) due to the presence of a gingival tumor that was initially diagnosed as granulation tissue by a dental surgeon. Examination of the patient's medical history revealed that a solid mass, measuring 1.5 cm in diameter, was identified in the right lobe of the liver 2 years prior to presentation at the current hospital; however, no biopsy was performed. Thus, the tumor was resected and histological examination resulted in an initial diagnosis of atypical squamous cell carcinoma. However, the histopathological characteristics, immunohistochemical features and serum α-fetoprotein expression levels supported a diagnosis of metastatic HCC. In conclusion, the present case study highlights the difficulties in diagnosing metastatic HCC without a history of primary HCC, and the importance of excluding a diagnosis of metastatic tumor when a lesion is identified in the gingival. Furthermore, it was determined that a final diagnosis of gingival metastasis of HCC predominantly depends on pathological characteristics and immunohistochemical features.

Dysplastic nodules with glypican-3 positive immunostaining: a risk for early hepatocellular carcinoma.

Glypican-3 (GPC3) has been reported to be a novel serum and histochemical marker for HCC. The positivity or negativity for GPC3 in hepatic precancerous lesions, such as dysplastic nodules (DN), has also been described. Moreover, our previous studies have demonstrated that some DN in liver cirrhosis represent monoclonal hyperplasia, and confirmed their neoplastic nature. However, additional studies must be performed to investigate further the relationship between DN with GPC3 positivity and HCC. Thus, we first investigated the expression of GPC3 in 136 HCC and 103 small DN (less than 1 cm in diameter) by immunohistochemical staining and determined the clonality of 81 DN from female patients using X-chromosome inactivation mosaicism and polymorphism of androgen receptor (AR) gene. Then we examined these samples for chromosomal loss of heterozygosity (LOH) at 11 microsatellite polymorphism sites. The results demonstrated that GPC3 immunoreactivity was detected in 103 of 136 HCC (75.7%) and 19 of 103 DN (18.4%), and the positive ratio correlated with HBsAg positivity. Clonality assays showed that 15 GPC3-positive DN from female patients, including 12 high-grade DN (HGDN), and 28 (42.4%) of 66 GPC3-negative DN, were monoclonal. In addition, among 19 GPC3-positive DN, chromosomal LOH was found at loci D6S1008 (100%, 19/19), D8S262 (52.6%, 10/19) and D11S1301 (57.9%, 11/19). However, the LOH frequency in GPC3-negative DN was 5.95% (5/84), 23.8% (20/84), and 4.76% (4/84) in three loci, respectively. Thus, we concluded that GPC3-positive DN, especially GPC3-positive HGDN, was really a late premalignant lesion of HCC.

[Genetic evolution analysis of matrix protein 2 gene of avian influenza H5N1 viruses from boundary of Yunnan province].

OBJECTIVE: To elucidate the variation in characterizations and genetic evolution of the matrix protein 2 or ion channel protein(M2) genes of avian influenza subtype H5N1 viruses in the boundary region of Yunnan province from 2008 to 2012. METHODS: A total of swab samples were collected from foreign poultry such as the junction between Yunnan and Vietnam, Laos,myanmar and wild birds in boundary region of Yunnan province from 2008 to 2012 and screened by H5N1 subtype-specific multiplex RT-PCR. The M genes of H5N1 virus from the positive samples were amplified by RT-PCR and cloned into pMD18-T vectors for sequencing. The alignment and phylogenetic analysis of M2 genes were performed with sequences of the known reference strains. RESULTS: A total of 71 positive samples were found out of 1240 samples and the positive rate was 5.72%. A total of 14 different M2 sequences were obtained from 30 positive samples and were divided into 3 distinct clades or sub-clades(1.2.1, 1.2.2 and 2) by phylogenetic analysis, 5, 7 and 2, respectively. The M2 genes and Hemagglutinin(HA) genes of H5N1 viruses from the boundary region of Yunnan province had showed different relationship of genetic evolution. The substitution or mutation of key amino acids sites had been found among the domains of epitope, adamantane-resistance, and poultry or human original viral strains. CONCLUSION: The M2 genes of H5N1 subtype viruses in boundary region of Yunnan province from 2008 to 2012 showed genetic divergence and the virus of clade 1.2.2 had become dominant epidemic strain in this region.

[Genetic evolution of non-structural gene among avian influenza H5N1 viruses isolated from the boundary of Yunnan province].

OBJECTIVE: To elucidate the characteristics of variation and the genetic evolution of non-structural protein (NS1, NS2) genes related to avian influenza subtype H5N1 viruses isolated from the boundary region of Yunnan province. METHODS: Swab samples were collected from foreign poultry and wild birds in the boundary regions of Yunnan province and screened by H5/N1 subtype-specific multiplex RT-PCR. The NS segment of H5N1 virus from the positive samples were amplified by RT-PCR and cloned into pMD18-T vectors for sequencing. The alignment and phylogenetic analysis on those available NS1, NS2 genes were performed with sequences of the known reference strains. RESULTS: 71 positive samples were identified from 1240 samples, with the positive rate as 5.72%. Fourteen different NS segment sequences were obtained from 30 representative positive samples and could be divided into 3 distinct clades or sub-clades (I-1, I-2 and II), by phylogenetic analysis. The NS1/NS2 genes and Hemagglutinin (HA) genes of H5N1 viruses from the boundary regions of Yunnan province showed different relationships regarding the characteristics on genetic evolution. The substitution or mutation of key amino acids sites had been noticed in the nuclear location signal domains, effect domain, and other pathogenicity markers. CONCLUSION: NS genes of H5N1 subtype viruses in boundary region of Yunnan province showed genetic divergence and the virus of clade I-2 and II had become dominant epidemic strains in this region since 2010.

Identification of genuine primary pulmonary NK cell lymphoma via clinicopathologic observation and clonality assay.

Extranodal natural killer (NK)/T-cell lymphoma, nasal type, is an uncommon lymphoma associated with the Epstein-Barr virus (EBV). It most commonly involves the nasal cavity and upper respiratory tract. Primary pulmonary NK/T cell lymphoma is extremely rare. If a patient with a NK or T-cell tumor has an unusual reaction to treatment or an unusual prognosis, it is wise to differentiate NK from T-cell tumors. The clinicopathologic characteristics, immunophenotype, EBV in situ hybridization, and T cell receptor (TCR) gene rearrangement of primary pulmonary NK cell lymphoma from a 73-year-old Chinese woman were investigated and the clonal status was determined using female X-chromosomal inactivation mosaicism and polymorphisms at the phosphoglycerate kinase (PGK) gene. The lesion showed the typical histopathologic characteristics and immunohistochemical features of NK/T cell lymphoma. However, the sample was negative for TCR gene rearrangement. A clonality assay demonstrated that the lesion was monoclonal. It is concluded that this is the first recorded case of genuine primary pulmonary NK cell lymphoma. The purpose of the present work is to recommend that pathologists carefully investigate the whole lesion to reduce the likelihood that primary pulmonary NK cell lymphoma will be misdiagnosed as an infectious lesion. In addition, TCR gene rearrangement and clonal analysis, which is based on female X-chromosomal inactivation mosaicism and polymorphisms at PGK and androgen receptor (AR) loci, were found to play important roles in differentiating NK cell lymphoma from T cell lymphoma. VIRTUAL SLIDES: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/5205300349457729.

[Isolation and characterization of rotavirus from bat].

Bats are considered as important animal reservoirs for many pathogenic viruses to humans. The viral metagenomic analysis was performed to study gut and lung tissues of 30 insectivorous bats collected in Yunnan Province and 26 reads were noted to group A rotavirus (RVA). Further RT-PCR screening on bat samples and in vitro viral isolation on cell cultures confirmed the presence of a novel RVA, named as RVA/Bat-tc/MYAS33/2013/G3P[10], in one of 30 Stoliczka's trident bats. The VP7 gene of this strain MYAS33 was closely related to that of an equine RVA strain from Argentina and the nucleotide sequence similarity was 93%, while its VP4 gene was a rare P[10] type and obtained the maximum sequence identity (94.8%) with that of a human strain from Thailand. The present study highlights the potential role of bats as reservoirs for RVAs.

Primary inflammatory myofibroblastic tumor of the breast with rapid recurrence and metastasis: A case report.

Primary inflammatory myofibroblastic tumor (IMT) of the breast is extremely rare; only 19 cases have been reported in the English literature. In the present study, we present a case of IMT in a 56-year-old female patient who was admitted to our hospital due to a mass found in her right breast. Mammogram and ultrasound revealed a well-circumscribed mass and surgery was performed. Histopathologically, the lesion was composed of spindle and inflammatory cells, including plasma cells and lymphocytes. Mitotic figures were not observed. Immunohistochemically, the tumor cells were positive for SM-actin, anaplastic lymphoma kinase (ALK) and vimentin and focal positive for desmin, but negative for NSE, S-100, CD117, CD34, NF, CD21, CD35 and CD68. Thus, we made a diagnosis of IMT and advised regular follow-up. However, the patient had local recurrence and metastasis to the left groin area 3, 7 and 10 months after the initial surgery. Notably, the histopathological characteristics of the recurrent and metastatic foci were similar to those of the initial specimen, but mitotic figures were clearly observed. Thus, we conclude that IMT shows occasionally malignant biological behavior although it is a neoplasm of intermediate biological potential that frequently recurs and rarely metastasizes. We advise that clinical physicians should regularly follow up patients after focal resection for IMT.

Gorham-Stout syndrome presenting in a 5-year-old girl with a successful bisphosphonate therapeutic effect.

Gorham-Stout syndrome (GSS), also known as Gorham-Stout disease, massive osteolysis, disappearing bone disease or phantom bone, is a rare disorder of the musculo-skeletal system. It most commonly involves the skull, shoulder and pelvic girdle. Histological examination reveals a progressive osteolysis always associated with an angiomatosis of blood vessels and sometimes of lymphatics, which seemingly is responsible for the destruction of the bone. It is extremely rare that Gorham-Stout syndrome involves the bones of the entire body. A 5-year-old girl complaining of intermittent and dull back pain for 3 months was admitted to a local hospital. X-ray revealed left pleural effusion, and the patient was diagnosed with tuberculous pleurisy. Thus, anti-tuberculosis therapy was performed. However, it was not effective. A soft mass with significant tenderness was found in the upper segment of the right leg 50 days afterwards. X-ray revealed multiple osteolysis of the bilateral clavicle, scapula, rib, vertebral body, ilium, sacrum, femur and tibia. The biopsy from the right tibia disclosed that the lesion was composed of hyperplastic blood vessels and fibrous tissues similar to hemangioma. Based on the above clinical, radiological and histopathological findings, the clinical physician confirmed a diagnosis of Gorham-Stout disease, and prescribed oral anti-osteoclastic medications consisting of bisphosphonates. At present, the girl is alive and healthy, and new lesions have not been noted.

Primary pulmonary malignant melanoma: a clinicopathologic study of two cases.

Malignant melanoma involving the respiratory tract is nearly always metastatic in origin, and primary tumors are very rare. To our knowledge, about 30 cases have been reported in the English literature, one of which involved multiple brain metastases. Here, we report two cases of primary pulmonary malignant melanoma. The first case, which occurred in a 52-year-old Chinese female patient who died 4 months after the initial diagnosis, involved rapid intrapulmonary and intracranial metastases. The second patient, a 65-year-old female, underwent surgical excision, and clinical examination, histopathological characteristics, and immunohistochemical features supported the diagnosis of pulmonary malignant melanoma. No evidence for recurrence and/or metastasis has been found more than one year after the initial surgery. To establish the diagnosis of primary pulmonary malignant melanoma, any extrapulmonary origin must be excluded by detailed examination. Moreover, the tumor should be removed surgically whether it occurs as a single lesion or multiple lesions. VIRTUAL SLIDE: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1480477335765055.

[Genetic diversification of avian influenza H5N1 virus in boundary areas of Yunnan province].

OBJECTIVE: To elucidate the genetic diversifications of avian influenza subtype H5N1 viruses in the boundary regions of Yunnan province during 2009 to July, 2011. METHODS: Swab samples were collected from foreign poultry and wild birds in boundary regions of Yunnan province during 2009 to July, 2011 and tested by H5/N1 subtype-specific multiplex RT-PCR. The HA genes of H5N1 virus from the positive samples were amplified by RT-PCR and cloned into pMD18-T vectors for sequencing. Both alignment and phylogenetic analysis were performed with sequences of the known reference strains. RESULTS: Fifteen different HA sequences were obtained from 36 representative positive samples and could be divided into 2 distinct Clades (2.3.2 and 2.3.4). Through phylogenetic analysis, Clade 2.3.2 and 2.3.4 could then be further divided into 3 (II-1 to II-3) and 2 smaller clades (I-1 and I-2), respectively. The viruses of Clade 2.3.2 II-1 and II-2 were new variant strains of H5N1 virus. The cleavage sites of HA from positive samples all possessed molecular characterization of highly pathogenic avian influenza virus. Mutation of key amino acids had been found among receptor binding sites, potential glycosylation sites, neutralizing epitopes and others. CONCLUSION: It seemed evident that the H5N1 subtype viruses showed genetic diversifications and had undergone the evolution progress of multi-clade (2.3.2, 2.3.4) to single calde (2.3.2) in the boundary regions of Yunnan province, during 2009 to July, 2011.