[Analysis of rare mutations associated with Thalassemia and their hematological characteristics in Chenzhou region of Hunan Province].

OBJECTIVE: To explore the distribution and hematological characteristics of rare thalassemia-associated mutations in Chenzhou region of Hunan Province with an aim to provide a basis for genetic counseling and effective prevention. METHODS: A total of 37 370 individuals enrolled from January 2015 to December 2021 were screened by routine blood test and hemoglobin electrophoresis. The genotypes were determined with high-throughput sequencing. RESULTS: A total of 8 455 thalassemia mutations (including 185 rare ones) were detected, which had involved 27 mutational types. Rare type α-Thalassemia --THAI and CD31 (AGG>AAG) have the typical microcytic hypochromic hematological features, whilst SEA-HPFH, CD14 (CTG>-TG), CD37 (TGG>TAG), -90(C>T), Codon 15 (G>A), IVS-I-128 (T>G), CD86 (GCC>GC-) and Chinese Gγ+(Aγδß)0 had typical microcytic hypochromic and ß-thalassemia-associated hematological features of elevated HbA2 or HbF. In addition, the -50（G>A）heterozygotes of ß-thalassemia had normal or slightly decreased MCV and MCH without an increase in HbA2. CONCLUSION: Various forms of thalassemia-associated mutations have been identified in the Chenzhou region of Hunan Province. Above finding has facilitated development of preventive and control strategies for thalassemia as well as birth health programs.

Enzymatic recombinase amplification coupled with CRISPR-Cas12a for ultrasensitive, rapid, and specific Porcine circovirus 3 detection.

Porcine circovirus type 3 (PCV3) is a disease associated with porcine dermatitis and nephrotic syndrome (PDNS) that has caused significant economic losses to swine herds since its discovery in 2016. To develop a simple, on-site, rapid, and sensitive assay to combat the spread of PCV3, we optimized the CRISPR/Cas12a (also known as Cpf1) system combined with enzymatic recombinase amplification (ERA) nucleic acid amplification to diagnose PCV3. The results showed that the ERA-CRISPR/Cas12a reaction could detect PCV3 within 1 h in genomic DNA harboring a minimum of seven copies. Additionally, we confirmed no cross-reactivity with PCV2, PCV4, or other porcine viruses, revealing the good specificity of this technique. These results demonstrated the ability of ERA-CRISPR/Cas12a to detect DNA at the single-molecule level and provide a rapid, simple, ultrasensitive, one-pot point-of-care test for PCV3 and suggest its potential for a variety of nucleic acid detection applications.

Application of vacuum sealing drainage to the treatment of seawater-immersed blast-injury wounds.

The aims of this study were to observe the effects of vacuum sealing drainage (VSD) with three different negative pressures on the wound healing rate, macrophage count and the expression of hyaluronic acid (HA) as well as its receptor CD44 in seawater-immersed blast-injury wounds (SIBIW) and to determine the optimal negative pressure value. In a minipig SIBIW model, different suction pressures and routine dressing were applied. Histological and immunohistochemical comparisons as well as molecular biology methods were performed to compare the wound healing conditions, macrophage count and the levels of HA and CD44. The wound healing rate of the VSD group was significantly higher than that of the control group, with the -120 mmHg group exhibiting the best effects. The macrophage count of the VSD group was higher than that of the control group. The HA level fluctuation was higher in the VSD group, with the -120 mmHg and the -180 mmHg groups showing the most significant fluctuation (P < 0·05). CD44 was expressed in the full-thickness wound-limbic tissues and was higher in the treatment group than that in the control group, with the -120 mmHg group having the most obvious expression. VSD significantly improved the healing ability and increased the macrophage count and the HA content. It also promoted CD44 expression. -120 mmHg is the optimal negative pressure value.

[Value of MR imaging in the diagnosis of intraductal papillary neoplasm of the bile duct].

OBJECTIVE: To analyze the value of MR imaging in diagnosis of intraductal papillary neoplasm of the bile duct (IPN-B). METHODS: Fourteen patients with intraductal papillary neoplasms of the bile duct confirmed by surgical pathology were included in this study. The patients underwent MR routine plain scanning and enhancement scanning (including T1WI, T2WI with fat suppression, FALSH T1WI, and three-phase enhancement scanning), diffusion weighted imaging(DWI) and magnetic resonance cholangiopancreatography (MRCP) before operation. The imaging data were reviewed and analyzed retrospectively in comparison with the surgical and pathological results. RESULTS: In these patients, 7 cases had tumors located in the left lobe, 2 cases had tumors in both the left and right lobes, 2 cases in the hepatic hilum, 2 cases in the common bile duct, and 1 case in both the right lobe and the common bile duct. Solitary or multiple intraductal masses could be found in 12 cases, with 11 cases appeared as papillary masses and one case as flat mass. In the other two cases the tumor was not visible (one case had too many stones, and in another case the tumor was too small). The tumors in the 12 cases showed hypointensity on T1WI and hyperintensity on T2WI. On the dynamic contrast-enhanced MRI, 11 cases showed mild and one showed moderate enhancement in arterial phase, and all the cases showed mildly and gradually delayed enhancement. On DWI, the lesion areas showed high signal intensity in all the cases, and the ADC value of the tumor area (1.697×10(-3)mm(2)/s) was significantly lower than that of the normal bile (3.973×10(-3)mm(2)/s) (t = -10.94, P < 0.05). Twelve cases demonstrated filling defects on primary MRCP coronary thin section images. On 3D-reconstruction MRCP images, 7 cases exhibited diffuse bile duct dilatation with the tumor areas more prominent, 3 cases exhibited aneurysmal bile dilatation, while the rest 4 cases exhibited segmental or lobar bile duct dilatation ( including 2 with invisible tumors ). In the 3 cases with aneurysmal bile dilatation, the multiple directions of MRCP images helped to find the communication between the aneurysmal dilatation and the bile duct. All the cases showed significant proximal bile duct dilatation (the extent of dilatation >100%), and 9 cases also showed distal bile duct dilatation. Bile duct stones were noted in 6 cases, 4 at the tumor area, and the other 2 away from the tumor area. No adjacent tissue invasion and no distal tissue or lymph node metastasis were observed. CONCLUSION: Intraductal papillary neoplasms of the bile ducts have characteristic imaging appearances on MRI, and MRI is an important method helpful to making correct diagnosis.

Four new furostanol saponins from the rhizomes and roots of Smilax scobinicaulis and their cytotoxicity.

Four new furostanol saponins 1-4, along with two known furostanol saponins 5 and 6 and one known spirostanol saponin 7 were isolated from the rhizomes and roots of Smilax scobinicaulis. The structures of the new saponins were elucidated as 26-O-ß-D-glucopyranoside-3ß,26-dihydroxy-(25R)-5α-furostan-22-methoxyl-6-one-3-O-α-L-arabinopyranosyl-(1→6)-ß-D-glucopyranoside (1), 26-O-ß-D-glucopyranoside-3ß,26-dihydroxy-(25R)-5α-furostan-22-methoxyl-6-one (2), 26-O-ß-D-glucopyranoside-3ß,26-dihydroxy-(25R)-5α-furostan-20(22)-en-6-one (3), 26-O-ß-D-glucopyranoside-3ß,23,26-trihydroxy-(23R, 25R)-5α-furostan-20(22)-en-6-one (4) on the basis of spectroscopic analysis. The isolated saponins were evaluated for cytotoxic activity against two human cancer cell lines including Hela (cervical carcinoma) and SMMC-7221 (hepatocellular carcinoma). Compounds 1 and 7 demonstrated cytotoxicity against the tested cell lines.

Ischemia and reperfusion-injured liver-derived exosomes elicit acute lung injury through miR-122-5p regulated alveolar macrophage polarization.

Acute lung injury (ALI) is a common postoperative complication, particularly in pediatric patients after liver transplantation. Hepatic ischemia-reperfusion (HIR) increases the release of exosomes (IR-Exos) in peripheral circulation. However, the role of IR-Exos in the pathogenesis of ALI induced by HIR remains unclear. Here, we explored the role of exosomes derived from the HIR-injured liver in ALI development. Intravenous injection of IR-Exos caused lung inflammation in naive rats, whereas pretreatment with an inhibitor of exosomal secretion (GW4869) attenuated HIR-related lung injury. In vivo and in vitro results show that IR-Exos promoted proinflammatory responses and M1 macrophage polarization. Furthermore, miRNA profiling of serum identified miR-122-5p as the exosomal miRNA with the highest increase in young rats with HIR compared with controls. Additionally, IR-Exos transferred miR-122-5p to macrophages and promoted proinflammatory responses and M1 phenotype polarization by targeting suppressor of cytokine signaling protein 1(SOCS-1)/nuclear factor (NF)-κB. Importantly, the pathological role of exosomal miR-122-5p in initiating lung inflammation was reversed by inhibition of miR-122-5p. Clinically, high levels of miR-122-5p were found in serum and correlated to the severity of lung injury in pediatric living-donor liver transplant recipients with ALI. Taken together, our findings reveal that IR-Exos transfer liver-specific miR-122-5p to alveolar macrophages and elicit ALI by inducing M1 macrophage polarization via the SOCS-1/NF-κB signaling pathway.

Anti-HIV and NO production inhibition activities of epi-aleuritolic acid derivatives.

Fifteen epi-aleuritolic acid derivatives were synthesized and evaluated for anti-HIV activity in 293 T cells and NO production inhibition activity. Of the derivatives, 1, 2, 3, 4, 11, and 13 showed relatively potent anti-HIV activity with EC50 values ranging from 5.80 to 13.30 µM. The most potent compound, 3α-2',2'-dimethylsuccinic acyl epi-aleuritolic acid (11), displayed significant anti-HIV activity with an EC50 value of 5.80 µM. Compounds 1, 3, 4, and 11 showed NO inhibition activity, with IC50 values ranging from 3.40 to 7.10 µM and compound 1 inhibited NO production with an IC50 value of 3.40 µM.

[Mechanism and action characteristics studies of a quinoxalinone compound against HIV-1 replication].

This study is to investigate the mechanism and action characteristics of 6-chloro-3-methyl-4-(2-methyoxycarbonylthiophene-3-sulfonyl)-3, 4-dihydroquinoxa-lin-2-(1 H)-one (XU07011) against HIV-1 replication. XU07011 anti-HIV activity was tested by using VSVG/HIV pseudotype viral system and confirmed by HIV-1 live viruses' infectious assay. Time of addition was used to test HIV-1 reverse transcription process. RNA-dependent DNA polymerase activity and RNase H activity were tested by using enzyme linked immunoabsorbent assay and fluorescence method. Wild type and nine NNRTIs-resistant reverse transcriptase enzymatic models and cell-based pharmacological models were used to evaluate XU07011 bio-characteristics. The results showed that XU07011 inhibited HIV-1 replication with IC50 of (0.057 +/- 0.01) micromol x L(-1) which was comparable to nevirapine [IC50: (0.046 +/- 0.01) micromol x L(-1)]. Mechanism study data indicated that XU07011 blocked HIV-1 reverse transcription process through acting on reverse transcriptase RNA-dependent DNA polymerase with IC 50 of (1.1 +/- 0.3) micromol x L(-1). The compound showed no effect on RNase H activity. XU07011 exhibited better activities comparing with nevirapine on K103N mutated NNRTIs-resistant HIV-1 strains. This study could provide a theoretical basis for novel anti-HIV reagents development.

Caspase 6/NR4A1/SOX9 signaling axis regulates hepatic inflammation and pyroptosis in ischemia-stressed fatty liver.

The mechanism of nonalcoholic fatty liver susceptibility to ischemia/reperfusion (IR) injury has not been fully clarified. Caspase 6 is a critical regulator in innate immunity and host defense. We aimed to characterize the specific role of Caspase 6 in IR-induced inflammatory responses in fatty livers. Human fatty liver samples were harvested from patients undergoing ischemia-related hepatectomy to evaluate Caspase 6 expression. in mice model, we generated Caspase 6-knockout (Caspase 6KO) mice to investigate cellular and molecular mechanisms of macrophage Caspase 6 in IR-stimulated fatty livers. In human liver biopsies, Caspase 6 expression was upregulated combined with enhanced serum ALT level and severe histopathological injury in ischemic fatty livers. Moreover, Caspase 6 was mainly accumulated in macrophages but not hepatocytes. Unlike in controls, the Caspase 6-deficiency attenuated liver damage and inflammation activation. Activation of macrophage NR4A1 or SOX9 in Caspase 6-deficient livers aggravated liver inflammation. Mechanistically, macrophage NR4A1 co-localized with SOX9 in the nuclear under inflammatory conditions. Specifically, SOX9 acts as a coactivator of NR4A1 to directly target S100A9 transcription. Furthermore, macrophage S100A9 ablation dampened NEK7/NLRP3-driven inflammatory response and pyroptosis in macrophages. In conclusion, our findings identify a novel role of Caspase 6 in regulating NR4A1/SOX9 interaction in response to IR-stimulated fatty liver inflammation, and provide potential therapeutic targets for the prevention of fatty liver IR injury.

Single-cell analysis reveals the immune heterogeneity and interactions in lungs undergoing hepatic ischemia-reperfusion.

Hepatic ischemia-reperfusion IR (HIR) is an unavoidable pathophysiological process during liver transplantation, resulting in systematic sterile inflammation and remote organ injury. Acute lung injury (ALI) is a serious complication after liver transplantation with high postoperative morbidity and mortality. However, the underlying mechanism is still unclear. To assess the phenotype and plasticity of various cell types in the lung tissue microenvironment after HIR at the single-cell level, single-cell RNA sequencing (scRNA-seq) was performed using the lungs from HIR-induced mice. In our results, we identified 23 cell types in the lungs after HIR and found that this highly complex ecosystem was formed by subpopulations of bone marrow-derived cells that signaled each other and mediated inflammatory responses in different states and different intervals. We described the unique transcriptional profiles of lung cell clusters and discovered two novel cell subtypes (Tspo+Endothelial cells and Vcan+ monocytes), as well as the endothelial cell-immune cell and immune cell-T cell clusters interactome. In addition, we found that S100 calcium binding protein (S100a8/a9), specifically and highly expressed in immune cell clusters of lung tissues and exhibited detrimental effects. Finally, the cellular landscape of the lung tissues after HIR was established, highlighting the heterogeneity and cellular interactions between major immune cells in HIR-induced lungs. Our findings provided new insights into the mechanisms of HIR-induced ALI and offered potential therapeutic target to prevent ALI after liver transplantation.

[Shizukaol F: a new structural type inhibitor of HIV-1 reverse transcriptase RNase H].

This study is to investigate the mechanism of action of lindenane disesquiterpenoid shizukaol F on HIV-1 replication. Real time quantity PCR, ELISA assay and fluorescence methods were used to test HIV-1 reverse transcription process, RNA-dependent DNA polymerase activity, and RNase H activity, respectively. It showed that shizukaol F inhibited LTR/Gag production of HIV-1 reverse transcription with an IC50 of 9.11 micromol x L(-1). This result is consistent with its inhibitory effect on HIV-1 replication (IC50 of 6.12 micromol x L(-1)). Mechanism studies showed that compound shizukaol F inhibited HIV-1 RT-RNase H with IC50 of 26.4 micromol x L(-1), but had no effect on HIV-1 RT RNA-dependent DNA polymerase activity. In conclusion, shizukaol F is a new structural type HIV-1 RNase H inhibitor. This discovery will provide a clue for new type of reverse transcriptase inhibitors development.

Study on the Classification Method of Rice Leaf Blast Levels Based on Fusion Features and Adaptive-Weight Immune Particle Swarm Optimization Extreme Learning Machine Algorithm.

Leaf blast is a disease of rice leaves caused by the Pyricularia oryzae. It is considered a significant disease is affecting rice yield and quality and causing economic losses to food worldwide. Early detection of rice leaf blast is essential for early intervention and limiting the spread of the disease. To quickly and non-destructively classify rice leaf blast levels for accurate leaf blast detection and timely control. This study used hyperspectral imaging technology to obtain hyperspectral image data of rice leaves. The descending dimension methods got rice leaf disease characteristics of different disease classes, and the disease characteristics obtained by screening were used as model inputs to construct a model for early detection of leaf blast disease. First, three methods, ElasticNet, principal component analysis loadings (PCA loadings), and successive projections algorithm (SPA), were used to select the wavelengths of spectral features associated with leaf blast, respectively. Next, the texture features of the images were extracted using a gray level co-occurrence matrix (GLCM), and the texture features with high correlation were screened by the Pearson correlation analysis. Finally, an adaptive-weight immune particle swarm optimization extreme learning machine (AIPSO-ELM) based disease level classification method is proposed to further improve the model classification accuracy. It was also compared and analyzed with a support vector machine (SVM) and extreme learning machine (ELM). The results show that the disease level classification model constructed using a combination of spectral characteristic wavelengths and texture features is significantly better than a single disease feature in terms of classification accuracy. Among them, the model built with ElasticNet + TFs has the highest classification accuracy, with OA and Kappa greater than 90 and 87%, respectively. Meanwhile, the AIPSO-ELM proposed in this study has higher classification accuracy for leaf blast level classification than SVM and ELM classification models. In particular, the AIPSO-ELM model constructed with ElasticNet+TFs as features obtained the best classification performance, with OA and Kappa of 97.62 and 96.82%, respectively. In summary, the combination of spectral characteristic wavelength and texture features can significantly improve disease classification accuracy. At the same time, the AIPSO-ELM classification model proposed in this study has sure accuracy and stability, which can provide a reference for rice leaf blast disease detection.

Rapid and Visual Detection of Porcine Parvovirus Using an ERA-CRISPR/Cas12a System Combined With Lateral Flow Dipstick Assay.

Porcine parvovirus (PPV) is one of the important causes of pig reproductive diseases. The most prevalent methods for PPV authentication are the polymerase chain reaction (PCR), enzyme-linked immunosorbent assay, and quantitative real-time PCR. However, these procedures have downsides, such as the fact that they take a long time and require expensive equipment. As a result, a rapid, visible, and economical clinical diagnostic strategy to detect PPV is necessary. In this study, three pairs of crRNA primers were designed to recognize the VP2 gene, and an ERA-CRISPR/Cas12a system for PPV detection was successfully developed. The approach involved isothermal detection at 37°C, and the method can be used for visual inspection. The detection limit of the ERA-CRISPR/Cas12a system was 3.75 × 102 copies/µL, and no cross reactions with other porcine viruses were found. In view of the preceding, a rapid, visible, and low-cost nucleic acid testing approach for PPV has been developed using the ERA-CRISPR/Cas12a system.

Lindenane disesquiterpenoids with anti-HIV-1 activity from Chloranthus japonicus.

Five new lindenane disesquiterpenoids, chlorajaponilides A-E (1-5), together with 11 known analogues were isolated from whole plants of Chloranthus japonicus. The structure and absolute configuration of 1 was confirmed by X-ray crystallography. Compounds 1 and 2 represent the first examples of lindenane disesquiterpenoids with a C-5 hydroxy group and a C-4-C-15 double bond. Compounds 8, 9, 11, and 12 showed anti-HIV-1 replication activities in both wild-type HIV-1 and two NNRTIs-resistant strains. Shizukaol B (8) exhibited the best activity against HIV(wt), HIV(RT-K103N), and HIV(RT-K103N) with EC50 values of 0.22, 0.47, and 0.50 µM, respectively. Compounds 8, 9, 11, and 12 had significant cytotoxicities against C8166 cells with CC50 values of 0.020, 0.089, 0.047, and 0.022, respectively, and exhibited inhibitory activities against HIV-1 with EC50 values of 0.0014, 0.016, 0.0043, and 0.0033 µM, respectively.

Identification and characterization of three novel small interference RNAs that effectively down-regulate the isolated nucleocapsid gene expression of SARS coronavirus.

Nucleocapsid (N) protein of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) is a major pathological determinant in the host that may cause host cell apoptosis, upregulate the proinflammatory cytokine production, and block innate immune responses. Therefore, N gene has long been thought an ideal target for the design of small interference RNA (siRNA). siRNA is a class of small non-coding RNAs with a size of 21-25nt that functions post-transcriptionally to block targeted gene expression. In this study, we analyzed the N gene coding sequences derived from 16 different isolates, and found that nucleotide deletions and substitutions are mainly located at the first 440nt sequence. Combining previous reports and the above sequence information, we create three novel siRNAs that specifically target the conserved and unexploited regions in the N gene. We show that these siRNAs could effectively and specifically block the isolated N gene expression in mammal cells. Furthermore, we provide evidence to show that N gene can effectively up-regulate M gene mediated interferon b (IFNb) production, while blocking N gene expression by specific siRNA significantly reduces IFNb gene expression. Our data indicate that the inhibitory effect of siRNA on the isolated N gene expression might be influenced by the sequence context around the targeted sites.

Association between Congenital Cytomegalovirus Infection and Brain Injury in Neonates: A Meta-analysis of Cohort Studies.

OBJECTIVE: To assess association between congenital cytomegalovirus (CMV) infection and brain injury in neonates. METHODS: The literatures from inception to November 4, 2020, were searched through PubMed, Embase, Cochrane Library, and Web of Science. Heterogeneity test was conducted for each indicator and measured by I 2 statistics. If I 2 ≥ 50%, the random effects model was applied; otherwise, the fixed effects model was used. Sensitivity analysis was performed for all models. Weighed mean difference (WMD) was used as the effect size for measurement data, and risk ratio (RR) was as the effect indicator. RESULTS: A total of 13 studies, including 4,262 congenital CMV infection neonates, were enrolled in this study. Our results showed that the rate of hearing impairment (RR: 2.105, 95% CI: (1.115, 3.971), P = 0.002), sensorineural hearing loss (SNHL) (RR: 17.051, 95% CI: (6.201, 46.886), P < 0.001), and microcephaly (RR: 2.283, 95% CI: (1.325, 3.935), P =0.003) in neonates infected congenital CMV was higher than that in control group. CONCLUSION: The risks of hearing impairment, SNHL, and microcephaly in neonates during childhood may be associated with congenital CMV infection. It is necessary to establish neonatal screening programs and comprehensive diagnostic tests for patients to reduce the risk of adverse brain damage to the congenital CMV infection as early as possible and to improve the prognosis of the newborn.

Acrocentric Chromosome Polymorphic Variants on Chinese Female Have Possible Association with Unexplained Recurrent Pregnancy Loss.

Recurrent pregnancy loss (RPL) occurs frequently in Chinese patients, but the causes for 40-50% RPL remain unexplained. This study aims to explore possible association between recurrent pregnancy loss and chromosome polymorphism. We performed a retrospective cohort study including a total number of 2967 Chinese patients from January in 2013 to December in 2019.These participants were divided into two groups: group I (RPL group) and group II (control group). Group I included 1791 patients who underwent RPL. Group II included 1176 individuals who underwent primary infertility. Their chromosome karyotype analyses were performed by Giemsa banding protocol. Pearson chi-squared test and Fisher's exact test were employed to evaluate the difference in frequency of chromosomal polymorphic variations between both groups. Chromosomal polymorphism occurred more frequently in RPL group compared with control group, the differences for 9qh+, inv(9), and Yqh+ between both groups were (χ2 = 6.697, p = 0.01), (χ2 = 6.242, p = 0.012), and (χ2 = 4.587, p = 0.032), respectively. The most significant difference of chromosomal polymorphism karyotype between RPL group and control group was found for acrocentric chromosome (D/G group) variants (χ2 = 30.7, p < 0.001). Moreover, the difference in frequency of D/G group variants between women and men in RPL group was remarkably significant (χ2 = 55.3, p < 0.001), and furthermore, D/G group variants occurred more frequently among women in RPL group than women in control group (χ2 = 44.3, p < 0.001). We found that acrocentric chromosomal polymorphism occurred preferentially on Chinese women with RPL. The fascinating finding provides us novel insight into the potential association of higher frequency of acrocentric chromosomal variations on Chinese women with unexplained RPL.

[Effects of organic manure partial substitution for chemical fertilizer on the photosynthetic rate, nitrogen use efficiency and yield of rice]. / æœ‰æœºè‚¥æ›¿ä»£éƒ¨åˆ†åŒ–è‚¥å¯¹æ°´ç¨»å ‰åˆé€ŸçŽ‡ã€æ°®ç´ åˆ©ç”¨çŽ‡å’Œäº§é‡çš„å½±å“.

Rational application of organic fertilizers is an effective approach to improve soil fertility, crop yield, and zero growth of chemical fertilizer in agricultural production. The rice variety 'Shennong9816' was planted in Shenyang, Liaoning Province, under seven different treatments: zero nitrogen fertilizer (CK), low nitrogen, 150 kg·hm-2(LN), medium nitrogen, 240 kg·hm-2 (MN), high nitrogen, 330 kg·hm-2(HN), medium nitrogen with replacement of chemical N by 10% organic manure (OMN10), medium nitrogen with replacement of chemical N by 20% organic manure (OMN20), and medium nitrogen with replacement of chemical N by 30% organic manure (OMN30). The effects of different treatments on photosynthetic rate, nitrogen absorption, nitrogen use efficiency, and yield were examined and the optimal fertilization scheme was determined. The results showed that the photosynthetic rate, biomass and yield were increased with the increases of nitrogen application rate, while the nitrogen use efficiency was decreased significantly. Compared with the MN treatment, the photosynthetic rate of OMN10 and OMN20 in filling stage was increased by 22.9% and 9.9%, respectively. The yield of OMN20 was increased by 3.8% compared to that of MN. The nitrogen agronomic efficiency and physiological efficiency of OMN20 were significantly improved by 8.1% and 13.3%, respectively. Moreover, the nitrogen agronomic efficiency and physiological efficiency of OMN20 were improved by 27.2% and 37.2% compared with the HN treatment. Thus, we concluded that the organic fertilizer replacement treatment could conserve soil fertility, achieve high yield and high nitrogen use efficiency, while reduce chemical nitrogen fertilizer application. The OMN20 treatment was recommended as a fertilizer application model due to its superior performance among the seven treatments.

Epidemiology and Evolution of Emerging Porcine Circovirus-like Viruses in Pigs with Hemorrhagic Dysentery and Diarrhea Symptoms in Central China from 2018 to 2021.

Porcine circovirus-like virus (PCLV) is a type of circular Rep-encoding single-stranded DNA virus and may be associated with the development of diarrheal symptoms in pigs. In this study, we retrospectively analyzed three years of past cases in Anhui, China, and reported a case of hemorrhagic enteritis and death in a pregnant sow possibly caused by PCLV. In addition, we analyzed the evolutionary characteristics of PCLV and found that mutation, recombination and selective pressure all played an important role in the evolution of PCLV. We identified N15D and T17S as well as L56T, T58R, K59Q, M62R, L75I and R190K mutations in two different branches, and we noted recombination events in the Rep of a group of Chinese strains. Analysis of selection pressure revealed that PCLV gained more positive selection, indicating that the virus is in a continuous evolutionary state. The PR2 plot, ENC-plot and neutrality analysis showed a greater role of natural selection than that of mutational pressure in the formation of codon usage patterns. This study is the first to identify PCLV in sows with hemorrhagic dysentery and death, and it provides new epidemiological information on PCLV infection in pigs in China.