Diversity of astroviruses in wild animals in Yunnan province, China.

BACKGROUND: Astroviruses (AstVs) are single-stranded RNA viruses that have been detected in a wide range of mammals and birds. They are associated with numerous interspecies transmissions and viral recombination events, posing a threat to human and animal health. METHODS: We collected 1,333 samples from wild animals, including bats, rodents, wild boars, and birds, from various states and cities in the Yunnan Province, China, between 2020 and 2023 to investigate the presence of AstVs. AstVs were detected using a polymerase chain reaction targeting the RdRp gene. Finally, the Molecular Evolutionary Genetics Analysis software was used to construct the phylogenetic tree. RESULTS: The overall positivity rate for AstVs was 7.12% in four species, indicating their widespread occurrence in the region. High genetic diversity among AstVs was observed in different animal species, suggesting the potential for interspecies transmission, particularly among rodents and birds. Additionally, we identified a novel AstV strain and, for the first time, provided information on the presence of bastroviruses in Yunnan, China. CONCLUSIONS: The widespread distribution and high genetic diversity of AstVs, along with the observed potential for interspecies transmission, highlight the importance of further investigation and surveillance in the region. The findings emphasize the need for increased attention to AstVs and their potential impact on human and animal health in Yunnan and other regions.

Comparison of clinicopathologic parameters and oncologic outcomes between type 1 and type 2 papillary renal cell carcinoma.

BACKGROUND: To compare the clinicopathologic parameters and oncologic outcomes between type 1 and type 2 papillary renal cell carcinoma (PRCC). METHODS: This study was approved by the review board (NO.XYFY2019-KL032-01). Between 2007 and 2018, 52 consecutive patients who underwent surgery at a single tertiary referral hospital were included. Clinicopathologic and survival data were collected and entered into a database. The Kaplan-Meier method, and univariate and multivariate Cox proportional hazard regression analyses were performed to estimate progression-free survival (PFS) and cancer-specific survival (CSS). RESULTS: Of the 52 patients, 24 (46.2%) were diagnosed with type 1 PRCC, and 28 (53.8%) had type 2 PRCC. The mean tumor size was 4.8 ± 2.5 cm. The two subtypes displayed different morphological features: foamy macrophages were more common in type 1 PRCC, while eosinophils and microvascular angiolymphatic invasion were more frequent in type 2 PRCC. Type 2 cases showed higher tumor stage and World Health Organization/International Society of Urological Pathology (WHO/ISUP) grade than type 1 cases (T3-T4: 43% vs 17%, P = 0.041; G3-G4: 43% vs 8%, P = 0.005). In univariate analysis, type 2 PRCC had a lower probability for PFS and CSS than patients with type 1 PRCC (P = 0.016, P = 0.049, log-rank test, respectively). In multivariate analysis, only WHO/ISUP grade (HR 11.289, 95% CI 2.303-55.329, P = 0.003) and tumor size (HR 1.244, 95% CI 1.034-1.496, P = 0.021) were significantly associated with PFS. CONCLUSIONS: PRCC subtype displayed different morphological features: foamy macrophages, eosinophils and microvascular angiolymphatic invasion are pathologic features that may aid in the distinction of the two subtypes. Histologic subtype of PRCC is not an independent prognostic factor and only WHO/ISUP grade and tumor size were independent predictors for PFS.

Overexpression of miR-122 promotes the hepatic differentiation and maturation of mouse ESCs through a miR-122/FoxA1/HNF4a-positive feedback loop.

BACKGROUND & AIMS: microRNA-122 is the only identified liver-specific miRNA and plays a crucial role in liver development, maintenance of hepatic homeostasis as well as tumourigenesis. In our previous differentiation of ESCs into hepatocytes, microRNA-122 (miR-122) was expressed at a relatively low level. Here, we aim to elucidate the effect and underlying mechanisms of miR-122 during differentiation of ESCs into hepatocytes. METHODS: Mouse ESCs were initially induced towards HPCs by activin A, FGF-4 and sodium butyrate and were subsequently transfected with a recombinant adenovirus expressing vector pAV.Ex1d-CMV>miR-122/IRES/eGFP 9 days after induction. Cells were analysed by real-time PCR, immunofluorescence, flow cytometry, microscopy and functional assays. Furthermore, microarray analysis was performed. RESULTS: We demonstrated that overexpression of miR-122 could effectively promote hepatic differentiation and maturation, as assessed by morphological and functional tests. The microarray analysis revealed that 323 genes were down-regulated, whereas 59 were up-regulated. Particularly, two liver-specific transcription factors, FoxA1 and HNF4a, were significantly up-regulated. Moreover, the expression of E-cadherin was dramatically increased and the proliferation of HPCs was suppressed, whereas knockdown of FoxA1 reduced E-cadherin expression and increased the proliferation of HPCs. In addition, the expression levels of FoxA1, HNF4a and E-cadherin in time-course transfection experiments with miR-122 were not significantly increased except in cells in which transfection with miR-122 occurred 9 days after induction. CONCLUSION: Overexpression of miR-122 at an appropriate stage could promote hepatic differentiation and maturation by regulating the balance between proliferation and differentiation, as well as the balance between EMT and MET, partially through a miR-122/FoxA1/HNF4a-positive feedback loop.

Genome Characterization of a Tailam Virus Discovered in Brown Rats in Yunnan Province, China.

Paramyxoviridae is one of the most well known and largest virus families, including some animal and human pathogens, such as the Hendra, Nipah, and Rinderpest viruses, with a high potential for the emergence of human diseases. Based on recent phylogenetic analyses, two new genera (Narmovirus and Jeilongvirus) have been described. The newly recognized genus Jeilongvirus has rapidly increased in number and has grown to 15 species from 7 a few years ago. However, little is known about the diversity, host range, or evolution of Jeilongvirus. As a well-known host reservoir for many pathogens, rodents have always been the focus for characterizing their pathogenic potential. In this study, we isolated a Tailam virus strain (RN-JH-YN-2022-1) belonging to the genus Jeilongvirus from Rattus norvegicus in Yunnan Province, China. The virus presented a near-complete genome (19,046 nucleotides). Similar to other members of the genus Jeilongvirus, the genome of RN-JH-YN-2022-1 contains eight basic genes (3'-N-P/V/C-M-F-SH-TM-G-L-5') with 88.88% sequence identity to Tailam virus (TL8K). Additionally, we discuss the pattern of genus Jeilongvirus diversity and the possible route of spread of the Tailam virus, which could provide new clues into the host range, virus diversity, and geographical distribution of the genus Jeilongvirus.

A Novel Moderate Constitutive Promoter Derived from Poplar (Populus tomentosa Carrière).

A novel sequence that functions as a promoter element for moderate constitutive expression of transgenes, designated as the PtMCP promoter, was isolated from the woody perennial Populus tomentosa. The PtMCP promoter was fused to the GUS reporter gene to characterize its expression pattern in different species. In stable Arabidopsis transformants, transcripts of the GUS reporter gene could be detected by RT-PCR in the root, stem, leaf, flower and silique. Further histochemical and fluorometric GUS activity assays demonstrated that the promoter could direct transgene expression in all tissues and organs, including roots, stems, rosette leaves, cauline leaves and flowers of seedlings and maturing plants. Its constitutive expression pattern was similar to that of the CaMV35S promoter, but the level of GUS activity was significantly lower than in CaMV35S promoter::GUS plants. We also characterized the promoter through transient expression in transgenic tobacco and observed similar expression patterns. Histochemical GUS staining and quantitative analysis detected GUS activity in all tissues and organs of tobacco, including roots, stems, leaves, flower buds and flowers, but GUS activity in PtMCP promoter::GUS plants was significantly lower than in CaMV35S promoter::GUS plants. Our results suggested that the PtMCP promoter from poplar is a constitutive promoter with moderate activity and that its function is presumably conserved in different species. Therefore, the PtMCP promoter may provide a practical choice to direct moderate level constitutive expression of transgenes and could be a valuable new tool in plant genetic engineering.

[The clinic research of the modified laparoscopic splenectomy for massive splenomegaly in the treatment of children with hematologic diseases].

OBJECTIVE: To summarize the experience and characteristics of the modified laparoscopic splenectomy for massive splenomegaly in the treatment of children with hematologic disease. METHODS: The clinical data of 30 cases of laparoscopic splenectomy for massive splenomegaly of children with hematologic disease from March 2007 to December 2011 was analyzed retrospectively. There were 18 male and 12 female patients, aging from 2 to 14 years. Primary disease included mediterranean anemia (17 cases), hereditary spherocytosis (4 cases) and idiopathic thrombocytopenic purpura (ITP, 9 cases). Dissection started with cutting off the gastrosplenic ligaments and lesser sac to fully reveal the splenic hilum, the splenic artery was clamped twice with 10 mm tiatanum clamp. When most of blood stored in the spleen back to heart through the veins and the splenic volume had already decreased, the splenic vein was ligated with 10 mm titanium clip and cut with ligsure and splenic pedicle separated. The Surgery and complication were recorded. For 1 week after surgery, the hemoglobin and platelet counts were reviewed. RESULTS: Twenty-six cases were performed successfully, and 4 cases were converted to open procedure. Of the 4 cases, 2 cases was obesity because of idiopathic thrombocytopenic purpura, 1 case was ß thalassaemia combined severe liver enlargement, and 1 case was after partial splenic embolization. In cases of laparoscopic splenectomy, operation time was 110 to 130 minutes, with an average of 120 minutes, and blood loss during operation was 35 to 180 ml, with an average of 45 ml. Compared with pre-operation, the hemoglobin of mediterranean anemia and hereditary spherocytosis patients were (92 ± 8) g/L, and blood platelet count of ITP patients was (127 ± 20)×10(9)/L, and they increased obviously at 1 week after operation (t = 4.175 and 8.253, both P = 0.000). CONCLUSION: The modified surgical method make the laparoscopic splenectomy for massive splenomegaly in many children with hematologic diseases possible, which was thought to be impossible in the past.

A novel signature constructed by differential genes of muscle-invasive and non-muscle-invasive bladder cancer for the prediction of prognosis in bladder cancer.

Background: Bladder cancer (BCa) is a malignant tumor that usually forms cancer cells in the inner lining of the bladder. Hundreds of thousands of people worldwide have BCa diagnosed each year. The purpose of this study was to construct a prognostic model by differential expression of genes between muscular and non-muscular invasive BCa, and to investigate the prognosis of BCa patients. Methods: The data of BCa patients was sourced from the GEO and TCGA database. Single-cell sequencing data was obtained from three patients in the GSE135337 database, and microarray data for verification was obtained from GSE32894. Univariate, Lasso and multivariate cox regression analyses were performed to construct the prognostic model. The prognostic features, immune features and drug sensitivity of the model were further evaluated. Single-cell data and microarray data were used to validate the differential expression of model genes between muscle-invasive and non-muscle-invasive BCa. The invasion and migration of BCa cells were evaluated using the transwell assay and wound-healing assay. The cell proliferation capacity was simultaneously evaluated using Colony formation experiments. The protein expression of the specific gene was detected by western blot analysis. Results: We identified 183 differentially expressed muscle-invasive-related differential genes (MIRDGs), among which four were selected to establish a prognostic model. Based on our signature, patients in different groups displayed varying levels of immune infiltration and immunotherapy profiles. Single-cell sequencing data and microarray data confirmed that four invasion-related genes were expressed at higher levels in muscle-invasive BCa. Given the critical role of S100A9 in the progression of BCa, we performed further analysis. The results showed that protein expression of S100A9 was high in muscle-invasive BCa, and S100A9 knockdown could inhibit the proliferation, migration and invasion of BCa. Conclusion: These findings demonstrated that the prognostic model for BCa patients was reasonably accurate and valid, and it may prove to be of considerable value for the treatment and prognosis of BCa patients in the future. S100A9 may become a better prognostic marker and potential therapeutic target to further guide clinical treatment decisions.

Selection of hepatocyte-like cells from mouse differentiated embryonic stem cells and application in therapeutic liver repopulation.

UNLABELLED: BACKGROUD/AIM: Because of the oncogenic risk, it is important to gain the homogeneous and purified cells from differentiated ESCs before transplantation. Here, we aim to select hepatocyte-like cells from differentiated ESCs, and investigate their growth, differentiation and neoplastic formation after intrahepatic transplantation. METHODS: Mouse ESCs were primarily induced by Dexamethesone, FGF-4 and HGF sequentially, then placed to a conditioning selection media consisting of 5% cholestatic sera and cultivated for 2 wks. After labeled by CFDA-SE, the selected cells were transplanted into mouse liver in therapeutic liver repopulation models. RESULTS: In the early stage of screening cultivation, most cells were suffered from apoptosis or even death. 1w later, some hepatocyte-like colony-forming units were observed, then the selected cells could grow and tend to be more mature, as assessed by morphological and functional tests. After intrahepatic transplantation, the labeled cells could proliferate and expressed albumin. Moreover, teratoma didn't form over 3 months. CONCLUSION: Our conditioning selection media could not only effectively select hepatocyte-like cells from differentiated ESCs, but further promote their growth and differentiation as well. After intrahepatic transplantation in therapeutic liver repopulation models, the selected cells could grow, differentiate and keep partial hepatic function. In particular, the transplantation was safe.

A modified laparoscopic splenectomy for massive splenomegaly in children with hematological disorder: a single institute retrospective clinical research.

PURPOSE: With the optimal acceptance of its clinical advantages, laparoscopic splenectomy (LS) emerged as a gold standard procedure as compared with open splenectomy (OS). However, it is still controversial and even counted as contraindication for massive splenomegaly. Here, we aim to summarize the experiences, characteristics and trends of modified LS for massive splenomegaly in children with hematological disorders. METHODS: Retrospective series of 57 pediatric patients with massive splenomegaly who underwent splenectomy from March 2007 to December 2011 were designated for this clinical analysis. The main outcome measures were dealt by statistics. For 30 cases of LS, we strictly adhered to the principle of using only three trocars to operate and initial ligation of the splenic artery, followed by retrieving the piecemeal of spleen through an accessory incision of 2-3 cm at 12 mm trocar port site. RESULTS: Of the 57 pediatric patients, 27 underwent OS and 30 underwent LS, respectively. Despite the operative time being shorter for OS than for LS (P < 0.001), the blood loss was lower in LS than in OS (P < 0.001); the time required for oral intake as well as duration of hospital stay was lower in LS than in OS (P < 0.001). Post-operatively, 7 (25.9 %) complications occurred in OS and 3 (10 %) in LS. The conversion rate of LS to OS was 13.33 % in four cases till 2009. CONCLUSIONS: Despite the conflicting reports regarding the safety of LS for massive splenomegaly, we demonstrated that our modified laparoscopic splenectomy in the treatment of children with massive splenomegaly in hematological diseases seemed to achieve the fundamental goal of less invasion; it was safe and feasible.

Tumor Location May Independently Predict Survival in Patients With M0 Squamous Cell Carcinoma of the Penis.

Background: To determine the association between tumor location and both clinicopathological characteristics and the survival of patients with M0 squamous cell carcinoma of the penis (SCCP). Methods: Data of 455 patients diagnosed with M0 SCCP between 1975 and 2018 were collected from the Surveillance, Epidemiology, and End Results (SEER) database of the United States National Cancer Institute. The effects of tumor location on overall survival (OS) and penile carcinoma-specific survival (PCSS) were analyzed using the Kaplan-Meier method. The Cox proportional hazards regression model was used to determine the impact of tumor location on PCSS. Results: SCCP was more likely to occur in the prepuce or glans (90%). Although no significant difference was observed between the OS of patients with M0 SCCP in the prepuce or glans and those with M0 SCCP in the body of the penis (p = 0.307), the former had better PCSS (p = 0.024). Moreover, M0 SCCP in the prepuce or glans was also significantly associated with better PCSS in patients with advanced age (age ≥ 60 years, p = 0.011), other ethnicities (p = 0.003), T2-T4 stage (p = 0.036), larger tumors (≥3 cm, p = 0.001), no regional lymph nodes removed (p = 0.044), and radical surgery (p = 0.027). Multivariate analysis confirmed that tumor location is an independent prognostic factor for patients with M0 SCCP [hazard ratio (HR) 1.881, p = 0.026]. Conclusions: Tumor location is an independent prognostic factor for patients with M0 SCCP, and tumors in the prepuce or glans portend better PCSS.

The clinical application value of miR-1269 as an unfavorable prognostic indicator of lung cancer.

BACKGROUND: Thanks to microRNAs (miR), a myriad of outstanding achievements have been made in multiple fields in recent years. miR-1269, a newly discovered miR, presents high expression profiles in lung cancer (LC), but its clinical implications in LC have not been clarified yet. METHODS: The miR-1269 expressions in the peripheral blood of LC patients, benign pulmonary disease (BPD) patients, and healthy controls were measured using qRT-PCR. Receiver operating characteristic (ROC) curves were employed for the identification of the diagnostic value of miR-1269 in LC, as were Kaplan-Meier (K-M) analyses and a Cox regression model to determine miR-1269's prognostic value in LC. RESULTS: qRT-PCR revealed higher miR-1269 expressions in the LC patients than in the BPD patients and the controls (P < 0.001). The LC patients with high miR-1269 expressions had advanced tumor stages (III-IV) and an increased probability of lymph node metastasis (LNM) (P < 0.01). Also, evidently elevated miR-1269 levels were observed in the peripheral blood of patients with the advanced tumor stages (III-IV) and LNM. Via ROC curves, we found that miR-1269 is of high clinical significance in the diagnosis of LC and advanced tumor stages. Our K-M survival analysis revealed a lowered 5-year survival rate in patients with high miR-1269 expressions, and our Cox regression analysis found that miR-1269 is an independent prognostic factor for LC. CONCLUSIONS: miR-1269, with high expression profiles in LC, indicates unfavorable patient prognoses, so it may be a viable diagnostic and prognostic indicator of LC.

Preoperative Neutrophil-to-Lymphocyte Ratio Was a Predictor of Overall Survival in Small Renal Cell Carcinoma: An Analysis of 384 Consecutive Patients.

OBJECTIVE: The aim of this study was to investigate the prognostic significance of the preoperative neutrophil-to-lymphocyte ratio (NLR) in small renal cell carcinoma (sRCC, ≤4 cm). METHODS: This study was approved by the review board (NO.XYFY2019-KL032-01). Between 2007 and 2016, a total of 384 consecutive patients who underwent curative surgery for sRCC at our institution were evaluated. Patients were divided into high NLR and low NLR groups by plotting the NLR receiver operating characteristic curve. The Kaplan-Meier method was utilized to graphically display survivor functions. Univariate and multivariate Cox proportional hazards regression analysis addressed time to overall survival (OS) and cancer-specific survival (CSS). RESULTS: Of the 384 patients, 264 (68.8%) were males and 120 (31.2%) were females. Median follow-up time after surgical resection was 54 months. One hundred and eighty-seven (48.7%) patients had a high NLR (≥1.97), and the remaining 197 (51.3%) had a low NLR (<1.97). Patients with high NLR were more likely to be aged compared with patients with low NLR (P=0.028). High NLR was associated with decreased OS and CSS compared with low NLR (P=0.028). High NLR was associated with decreased OS and CSS compared with low NLR (P=0.028). High NLR was associated with decreased OS and CSS compared with low NLR (P=0.028). High NLR was associated with decreased OS and CSS compared with low NLR (. CONCLUSIONS: Elevated preoperative NLR is an independent adverse prognostic factor for OS after surgery with curative intent for sRCC.

[In vitro effect of medicinal scorpion on the larvae of Ancylostoma caninum].

Five gram stool sample from dog infected with Ancylostoma caninum was smeared on filter paper for routine cultivation under 35 degrees C with moist condition for 24 hours. Hookworm larvae were separated and divided into 3 bottles through precipitation-washing method. Traditional Chinese medicine scorpion (2 ml), albendazole (10 mg) and normal saline (2 ml) (control) were added respectively into the 3 bottles, which were cultured for another 24 hours. The growth and development of the larvae were observed under microscope. In scorpion group, the larva body shrank, development stopped, and with an unclear internal structure. Treated with albendazole, the larvae became rigid and shriveled with rough body surface and vague internal structure. The results indicate that both medicinal scorpion and albendazole have considerable effect in inhibiting the development of hookworm larvae and the effect of albendazole is stronger.

Purulent meningitis caused by Rhodococcus equi: A case report.

RATIONALE: Purulent meningitis refers infection of the subarachnoid space by various purulent bacteria and the corresponding inflammation of the leptomeninges. However, purulent meningitis due to Rhodococcus equi is extremely rare. PATIENT CONCERNS: A 40-year-old man presented with fever and intermittent headache for 6 days. Two hours prior to admission, he developed epileptic seizures. DIAGNOSES: Brain computed tomography and magnetic resonance imaging showed intracerebral malacic lesions. Bacterial culture of cerebrospinal fluid revealed the presence of R. equi. A diagnosis of purulent meningitis caused by R. equi was made. INTERVENTIONS: The patient was treated with intravenous meropenem (1000 mg every 8 hours) for 19 days; then he was discharged and instructed to continue the intravenous meropenem for two weeks. After a follow-up period of 2 months, the patient had recovered completely. OUTCOMES: After a follow-up period of 2 months, the patient had recovered completely. LESSONS: Central nervous system infection caused by R. equi is rare. Early bacterial culture of CSF is important for timely diagnosis. With sufficient antibiotic therapy, the prognosis can be favorable.

Analysis of treatment of large abdominal malignancies in children complicated with abdominal compartment syndrome: Report of six cases.

To explore effective treatment of large abdominal malignancies in children complicated with abdominal compartment syndrome (ACS).Six children with large abdominal malignancies complicated with ACS were admitted to our department from January 2013 to January 2016, and the changes in their breathing, heart rate, oxygen saturation, abdominal circumference, bladder pressure, and urine output, as well as the treatment measures and outcomes, were retrospectively analyzed.The 6 children included 1 child with bilateral nephroblastoma, 1 child with abdominal alveolar rhabdomyosarcoma, 1 child with right ovarian malignant teratoma complicated with abdominal glioma, 1 child with abdominal malignant teratoma, 1 child with right nephroblastoma, and 1 child with left adrenal gland neuroblastoma. All patients were treated in a timely manner. The first 4 children underwent abdominal cavity decompression through surgical resection of the tumor, and the ACS was successfully cured allowing for follow-up care, whereas the last 2 patients failed to receive emergency surgery and eventually died due to the gradual aggravation of ACS.Decompression through surgical resection of the tumor is the only effective measure for treating large abdominal malignancies in children complicated with ACS.

[Multi-scale correlation analysis of soil organic carbon with its influence factors using wavelet transform].

Based on GIS, this paper chose the soil organic carbon (SOC) density in soil surface layer (0-20 cm) and its influence factors (NDVI, elevation, slope and aspect) as research objects, one-dimensional discrete wavelet transform (DWT) was used as the multi-scale decomposition tool to quantitatively revealed the multi-scale correlation relationships among SOC density and its influence factors on the grid scale along 4 transects of the mountainous area of Guangdong Province. The results showed that the correlation among SOC density and its influence factors was scale-dependent with varying degree. The influence of NDVI was strongest at the scales of 2, 8 and 16 km, while elevation showed its greatest influence at the scales of 8 and 16 km. The control action of slope was rather weak, with a less significant correlation depending on scale. The negative effect of aspect became stronger with increasing scale at > 2 km scale. The SOC density of the different transects was affected by various factors, of which NDVI and elevation were the main factors, and slope and aspect only reacted with individual transects at larger scales.

[Establishment of microarray for detecting mutation in HBV pre-core/core and basic core promoter regions].

OBJECTIVE: To develop a sensitive and specific microarray for detecting mutations of HBV pre-core/core and basic core promoter regions in the clinic. METHODS: Site-specific oligonucleotide probes were designed and immobilized to microarray slides and hybridized to HBV gene fragments amplified with specific biotin-labeled primer using asymmetrical PCR. The specificity and sensitivity of the method were estimated. And the microarray was applied to detect 138 clinical serum samples with HBV-DNA. RESULTS: The mutations of HBV pre-core/core and basic core promoter regions can be specifically detected using the microarray, and the sensitivity was 1 x 10(1) copies/microl. Among 138 samples, 40 samples had T1762/ A1764 mutation, 11 samples had C1814 mutation, and 16 samples had A1896 mutation. The A1896 mutation rate in high HBV-DNA load group was significantly higher than that in low HBV-DNA load group (P < 0.01). CONCLUSION: An DNA microarray assay was successfully established to detect the mutations in HBV pre-core/core and basic core promoter regions. The A1896 mutation in pre-core/core region maybe involve in duplication of HBV.