A Zika virus envelope mutation preceding the 2015 epidemic enhances virulence and fitness for transmission.

Arboviruses maintain high mutation rates due to lack of proofreading ability of their viral polymerases, in some cases facilitating adaptive evolution and emergence. Here we show that, just before its 2013 spread to the Americas, Zika virus (ZIKV) underwent an envelope protein V473M substitution (E-V473M) that increased neurovirulence, maternal-to-fetal transmission, and viremia to facilitate urban transmission. A preepidemic Asian ZIKV strain (FSS13025 isolated in Cambodia in 2010) engineered with the V473M substitution significantly increased neurovirulence in neonatal mice and produced higher viral loads in the placenta and fetal heads in pregnant mice. Conversely, an epidemic ZIKV strain (PRVABC59 isolated in Puerto Rico in 2015) engineered with the inverse M473V substitution reversed the pathogenic phenotypes. Although E-V473M did not affect oral infection of Aedes aegypti mosquitoes, competition experiments in cynomolgus macaques showed that this mutation increased its fitness for viremia generation, suggesting adaptive evolution for human viremia and hence transmission. Mechanistically, the V473M mutation, located at the second transmembrane helix of the E protein, enhances virion morphogenesis. Overall, our study revealed E-V473M as a critical determinant for enhanced ZIKV virulence, intrauterine transmission during pregnancy, and viremia to facilitate urban transmission.

Repair mechanism of Yishen Tongluo formula on mouse sperm DNA fragmentation caused by polystyrene microplastics.

CONTEXT: Plastics can break down into millions of microplastic (MPs, < 5 mm) particles in the soil and ocean. These MPs can then affect the function of the reproductive system. There is currently no effective solution to this problem aside from traditional Chinese medicine. We have previously used Yishen Tongluo formula (YSTL) to treat sperm DNA damage caused by some toxic substances. OBJECTIVE: To investigate the mechanism underlying the repair of mouse sperm DNA fragmentation caused by polystyrene microplastics by YSTL. MATERIALS AND METHODS: An animal model of polystyrene microplastic (PS-MP)-induced sperm DNA damage was replicated by gavage of SPF ICR (CD1) mice PS-MPs at 1 mg/d and treated with YSTL at 11.89, 23.78 and 47.56 g/kg, respectively, for 60 days. The Sperm DNA fragmentation index (DFI) of each group was detected and compared. The target genes of YSTL identified by transcriptomic and proteomic analyses were validated by qRT-PCR and western blotting. RESULTS: The DFI of the PS group (20.66%) was significantly higher than that of the control group (4.23%). The medium and high doses of the YSTL group (12.8% and 11.31%) exhibited a significant repairing effect. The most enriched pathway was PI3K/Akt. TBL1X, SPARC, hnRNP0, Map7D1, Eps8 and Mrpl27 were screened and SPARC was validated. DISCUSSION AND CONCLUSIONS: The precise mechanism by which YSTL inhibits PD-MPs DNA damage may be associated with the PI3K/Akt pathway and SPARC. It provides a new direction for using traditional Chinese medicine to prevent and repair reproductive system injury caused by MPs.

[Effect of bird-induced anus-lifting surgery combined with oral Relinqing Granules on LUTS/CP with damp heat syndrome: Evaluation by infrared thermal imaging].

OBJECTIVE: To observe the clinical effect of bird-induced anus-lifting surgery (BIAL) combined with oral Relinqing Granules (RG) in the treatment of lower urinary tract symptoms / chronic prostatitis (LUTS/CP) with damp-heat syndrome using infrared thermal imaging (ITI). METHODS: This study included 165 cases of LUTS/CP with damp-heat syndrome treated in the Departments of Andrology and Urology of our hospital, which were divided into three groups of an equal number and treated by medication of oral tamsulosin hydrochloride at 0.2 mg, qd plus levofloxacin at 0.2 g bid (control), anus-lifting exercise (15 lifts once a day) combined with oral RG at 8 g tid (routine treatment) and BIAL (15 lifts once a day) combined with oral RG at 8 g tid (BIAL+RG), respectively, all for two weeks. We recorded the ITI indicators, NIH-CPSI and Self-Rating Anxiety Scale (SAS) scores of patients before and after treatment, observed adverse reactions during the treatment, and compared them among the three groups of patients. RESULTS: Compared with the baseline, NIH-CPSI and SAS scores were significantly reduced in all the three groups after treatment (P < 0.05), even more significantly in the BIAL+RG (10.49 ± 2.97 and 49.73 ± 2.79) than in the control (21.15 ± 3.56 and 52.62 ± 5.38) and routine treatment groups (15.55 ± 2.77 and 52.35 ± 3.87) (P < 0.05). The ITI temperature at the related parts of the body was remarkably decreased in the three groups after treatment (P < 0.05), even lower in the BIAL+RG group (30.47 ± 1.61, 32.14 ± 1.25, 31.16 ± 1.58, 31.97 ± 1.19) than in the control (33.39 ± 0.6, 33.64 ± 1.07, 32.78 ± 0.58, 33.18 ± 0.78) and routine treatment groups (31.93 ± 1.14, 33.06 ± 0.8, 33.08 ± 0.82, 33.17 ± 1.05) (P < 0.05), with statistically significant difference from the baseline in the BIAL+RG group after 12 hours and 24 hours of treatment (ï¼»34.26 ± 0.06ï¼½ vs ï¼»32.65 ± 0.05ï¼½ and ï¼»32.72 ± 0.09ï¼½, P < 0.05), but not between the two time points (P > 0.05). The incidence rate of adverse reactions was markedly lower in the BIAL+RG and routine treatment groups than in the control (P < 0.05) but not significantly different between the BIAL+RG and routine treatment groups (P > 0.05). CONCLUSION: Bird-induced anus-lifting surgery combined with oral Relinqing Granules can safely and significantly improve infrared thermal imaging indicators and NIH-CPSI scores in LUTS/CP patients with damp-heat syndrome.

Repair mechanism of Wuwei Fuzheng Yijing formula in di-2-ethylhexyl phthalate-induced sperm DNA fragmentation in mice.

CONTEXT: Di-2-ethylhexyl phthalate (DEHP), a known persistent organic pollutant, can increase the sperm DNA fragmentation index (DFI). OBJECTIVE: To investigate the mechanism underlying the repair of DEHP-induced sperm DNA damage in mice by Wuwei Fuzheng Yijing (WFY) formula. MATERIALS AND METHODS: The potential targets of WFY and sperm DNA fragment (SDF) were obtained from the TCMSP, BATMAN-TCM, OMIM and GeneCards. The protein-protein interaction (PPI) network, GO and KEGG pathway analyses of WFY-SDF were constructed. An animal model of DEHP-induced sperm DNA damage was replicated by gavage of SPF ICR (CD1) mice DEHP at 1 g/kg/d and treated with WFY at 8.92, 17.84 and 35.67 g/kg, respectively, for 60 d. Sperm DFI of each group was detected and compared. The target genes of WFY identified by transcriptomic and proteomic analyses were validated by qRT-PCR and Western blotting. RESULTS: Network pharmacology pathway analysis indicated that PI3K/Akt was the potential target of WFY on SDF. The DFI of the DEHP group (25.48%) was significantly higher than that of the control group (4.02%). The high-dose WFY group (19.05%) exhibited the most significant repairing effect. The related pathways were PI3K/Akt and metabolic. Aass, Aldh1a7, GSTA3, betaine homocysteine S-methyltransferase (Bhmt), Mug2 and Svs1 were screened and Bhmt was validated. DISCUSSION AND CONCLUSIONS: WFY can repair sperm DNA damage caused by DEHP, and the mechanism may be related to PI3K/Akt and metabolic pathways, and Bhmt. This provides a new direction for using traditional Chinese medicine to prevent and repair reproductive system injury caused by pollutants.

[Huayu Jiedu Shengjing Decoction improves semen quality and reduces TCM syndrome scores in patients with varicocele-induced asthenospermic infertility].

OBJECTIVE: To observe the clinical efficacy of "Huayu Jiedu Shengjing Decoction" (HJSD) in the treatment of varicocele (VC)-induced asthenospermic infertility and its action mechanism. METHODS: Using computer-generated random numbers, we equally divided 88 patients meeting the study criteria into an experimental and a control group, the former treated orally with HJSD plus or minus, while the latter with Maizhiling Tablets and levocarnitine, both for a course of 12 weeks. After medication, we obtained TCM syndrome scores, sperm motility, sperm DNA fragmentation index (DFI), the seminal cord venous ultrasonographic index, and levels of nitric oxide (NO), reactive oxygen species (ROS) and superoxide dismutase (SOD) in the seminal plasma from the patients, compared the therapeutic effects between the two groups, and analyzed the correlation among the obtained parameters. RESULTS: The total effectiveness rate was dramatically higher in the experimental than in the control group (86.04% vs 73.74%, P < 0.01). The TCM syndromes scores, sperm motility, sperm DFI, and seminal plasma NO, ROS and SOD were all more significantly improved in the former than in the latter group (P < 0.05). CONCLUSION: Huayu Jiedu Shengjing Decoction can improve semen quality and reduce TCM syndrome scores without adverse reactions in patients with VC-induced asthenospermic infertility, which may be attributed to its effect of improving antioxidation and local blood flow.

[Yishen Tongluo Prescription for repairing benzo(a)pyrene-induced sperm DNA damage in male rats: Effect and mechanism].

OBJECTIVE: To investigate the repairing effect of Yishen Tongluo Prescription (YTP) on sperm DNA fragmentation index (DFI) in male rats exposed to benzo(a)pyrene (BaP) and its possible mechanism. METHODS: Thirty Wistar male rats were equally randomized into a blank control, a BaP-exposure and a YTP intervention group, those in the latter two groups exposed to BaP at 20 mg/kg/d for 60 consecutive days, and those in the YTP intervention group treated intragastrically with YTP from the 31st day of BaP exposure for a total of 30 days. After the last administration, the sperm DFI of the rats was detected by sperm chromatin structure analysis, the levels of FSH, LH and T in the serum and superoxide dismutase (SOD), malondialdehyde (MDA), nitric oxide (NO) and adenosine triphosphate (ATP) in the testis were measured by ELISA. RESULTS: Compared with the blank controls, the rats in the BaP-exposure group showed significantly increased DFI ( ï¼»4.23 ± 1.40ï¼½% vs ï¼»12.46 ± 3.07ï¼½%, P < 0.05), serum FSH (ï¼»1.76 ± 0.31ï¼½ vs ï¼»2.53 ± 0.28ï¼½ U/L, P < 0.05) and LH (ï¼»30.59 ± 2.14ï¼½ vs ï¼»39.72 ± 2.80ï¼½ U/L, P < 0.05), decreased levels of serum T (ï¼»5.33 ± 0.43ï¼½ vs ï¼»4.42 ± 0.38ï¼½ nmol/L, P < 0.05) and testicular SOD (ï¼»166.18 ± 3.74ï¼½ vs ï¼»113.23 ± 10.76ï¼½ U/ml, P < 0.05) and ATP (ï¼»41.23 ± 2.21ï¼½ vs ï¼»33.48 ± 2.74ï¼½ mol/L, P < 0.05), and elevated contents of MDA (ï¼»7.55 ± 0.93ï¼½ vs ï¼»10.59 ± 1.17ï¼½ nmol/ml, P < 0.05) and NO (ï¼»44.23±4.47ï¼½ vs ï¼»54.49 ± 3.13ï¼½ mol/L, P < 0.05). All the above parameters returned to normal after YTP intervention (DFI: ï¼»5.73 ± 2.46ï¼½%, FSH: ï¼»2.07 ± 0.45ï¼½ U/L, LH: ï¼»33.94 ± 4.44ï¼½ U/L, T: ï¼»4.96 ± 0.24ï¼½ nmol/L, SOD: ï¼»135.22 ± 7.26ï¼½ U/ml, ATP: ï¼»38.26 ± 2.14ï¼½ mol/L, MDA: ï¼»8.37 ± 1.29ï¼½ nmol/ml, NO: ï¼»48.36 ± 3.98ï¼½ mol/L), with statistically significant difference from those in the BaP-exposure group (all P < 0.05). CONCLUSION: Yishen Tongluo Prescription can repair BaP-induced sperm DNA damage in male rats, which may be attributed to its effects of suppressing oxidative damage.

One-step synthesis of fluorescence-enhanced carbon dots for Fe (III) on-off-on sensing, bioimaging and light-emitting devices.

Carbon quantum dots (CDs), as one of the most potential fluorescent sensing materials, have attracted lots of attention in recent years. However, the low quantum yields, complicated separation and purification procedures have limited its application and large-scale production. In this paper, a facile and universal method was successfully developed to synthesize CDs-silica (SiO2) spheres composites (CS composites), which not only demonstrates 10-fold fluorescence enhancement compared with single CDs but also enables the purification with a simple centrifugation step. Meanwhile, fabricated composites also presented superior photoluminescence (PL) stability and high PL intensity, even in some tough environments such as acid/alkali aqueous solution, high/low temperature, and high ion concentration aqueous solution. This simple, fast, low-cost, and efficient synthesis method would potentially expand the application of CDs for clinical analysis, optical sensing (ferric ion (Fe3+) and pyrophosphate), bioimaging and light-emitting diodes (LEDs).

Water-stable perovskite-on-polymer fluorescent microspheres for simultaneous monitoring of pH, urea, and urease.

Perovskite materials have attracted attention due to their excellent optical and electrical properties; however, their unsatisfactory stability limits their application in biochemical detection. In this paper, CsPbBr3 perovskite quantum dots were successfully encapsulated in poly(styrene/acrylamide) microspheres, using a swelling-shrinking method. The manufactured perovskite microspheres (PDPS composites) not only maintained strong photoluminescence (PL) stability but also demonstrated great water solubility. Additionally, a real-time pH monitoring platform was constructed based on the prepared PDPS composites and dopamine, and the system showed a good linear relationship in a pH range of 4-12. Furthermore, urea could be hydrolyzed to produce hydroxyl groups, thereby increasing the pH of the solution. Therefore, this system was then extended for urea and urease detection. As a result, the detection limits of urea and urease were recorded as 1.67 µM and 2.1 mU/mL, respectively. This development provides an interesting demonstration of the expanding list of applications of perovskite materials.

Chikungunya Virus: Role of Vectors in Emergence from Enzootic Cycles.

Chikungunya virus (CHIKV), a re-emerging mosquito-borne arbovirus, has caused millions of cases of severe, often chronic arthralgia during recent outbreaks. In Africa, circulation in sylvatic, enzootic cycles involves several species of arboreal mosquito vectors that transmit among diverse nonhuman primates and possibly other amplifying hosts. Most disease occurs when CHIKV emerges into a human-amplified cycle involving Aedes aegypti and sometimes Aedes albopictus transmission and extensive spread via travelers. Epidemiologic studies suggest that the transition from enzootic to epidemic cycles begins when people are infected via spillover in forests. However, efficient human amplification likely only ensues far from enzootic habitats where peridomestic vector and human densities are adequate. Recent outbreaks have been enhanced by mutations that adapt CHIKV for more efficient infection of Ae. albopictus, allowing for geographic expansion. However, epistatic interactions, sometimes resulting from founder effects following point-source human introductions, have profound effects on transmission efficiency, making CHIKV emergence somewhat unpredictable.

[Luobufukebiri Pills improve semen quality and seminal plasma-related biochemical indexes in patients with asthenospermia].

OBJECTIVE: To investigate the clinical effect of Luobufukebiri Pills (LP) on asthenospermia and its action mechanism. METHODS: A total of 206 patients with asthenospermia in Henan Provincial Hospital of Traditional Chinese Medicine from March 2018 to December 2018 were randomly assigned to receive oral LP (the experimental group, n = 103) bid 10 pills each time or vitamin E soft capsules (the control group, n = 103) bid 100 mg each time for a course of 12 weeks. We compared the semen volume, sperm concentration, sperm motility, levels of α-glucosidase (α-Glu), fructose (Fru), malondialdehyde (MDA) and superoxide dismutase (SOD) in the seminal plasma, and the number of pregnancies between the two groups of patients before and after treatment. RESULTS: All the patients finished the experiment except 1 in the control group due to personal reasons. After 12 weeks of medication, the patients in the experimental group showed significant increases over the baseline in sperm concentration (ï¼»36.27 ± 11.05ï¼½ vs ï¼»25.96 ± 8.63ï¼½ ×106/ml, P < 0.05), grade a sperm (ï¼»16.92 ± 5.72ï¼½% vs ï¼»9.34 ± 3.18ï¼½%, P < 0.05), grade a+b sperm (ï¼»36.08 ± 6.84ï¼½% vs ï¼»19.88 ± 4.65ï¼½%, P < 0.05), total sperm motility (ï¼»50.37 ± 10.38ï¼½% vs ï¼»24.62 ± 6.12ï¼½%, P < 0.05), and the levels of α-Glu (ï¼»46.94 ± 9.16ï¼½ vs ï¼»32.25 ± 8.30ï¼½ U/ml, P < 0.05), Fru (ï¼»28.08 ± 5.72ï¼½ vs ï¼»16.38 ± 4.62ï¼½ µmol/ml, P < 0.05) and SOD (ï¼»182.28 ± 18.44ï¼½ vs ï¼»100.37 ± 10.22ï¼½ U/ml, P < 0.05), but a decrease in the content of seminal plasma MDA (ï¼»10.37 ± 2.93ï¼½ vs ï¼»15.62 ± 4.68ï¼½ nmol/ml, P < 0.05). Increases were also observed in the control group over the baseline in grade a sperm (ï¼»12.40 ± 6.33ï¼½% vs ï¼»9.26 ± 2.94ï¼½%, P < 0.05), grade a+b sperm (ï¼»25.42 ± 5.24ï¼½% vs ï¼»19.43 ± 4.17ï¼½%, P < 0.05), total sperm motility (ï¼»40.22 ± 9.72ï¼½% vs ï¼»23.99 ± 5.77ï¼½%, P < 0.05), and levels of α-Glu (ï¼»40.31 ± 8.22ï¼½ vs ï¼»31.06 ± 7.51ï¼½ U/ml, P < 0.05) and SOD (ï¼»139.71 ± 14.08ï¼½ vs ï¼»102.11 ± 9.24ï¼½ U/ml, P < 0.05), but a decrease in the content of seminal plasma MDA (ï¼»12.72 ± 3.11ï¼½ vs ï¼»15.62 ± 4.68ï¼½ nmol/ml, P < 0.05). The total clinical effectiveness rate was remarkably higher in the experimental than in the control group (87.38% vs 58.82%, P < 0.05). CONCLUSIONS: Luobufukebiri Pills are safe and effective for the treatment of asthenospermia, which may be attributed to their ability of improving oxidative stress damage.

ICTV Virus Taxonomy Profile: Togaviridae.

The Togaviridae is a family of small, enveloped viruses with single-stranded, positive-sense RNA genomes of 10-12 kb. Within the family, the genus Alphavirus includes a large number of diverse species, while the genus Rubivirus includes the single species Rubella virus. Most alphaviruses are mosquito-borne and are pathogenic in their vertebrate hosts. Many are important human and veterinary pathogens (e.g. chikungunya virus and eastern equine encephalitis virus). Rubella virus is transmitted by respiratory routes among humans. This is a summary of the International Committee on Taxonomy of Viruses (ICTV) Report on the taxonomy of the Togaviridae, which is available at www.ictv.global/report/togaviridae.

Comprehensive Genome Scale Phylogenetic Study Provides New Insights on the Global Expansion of Chikungunya Virus.

Since the India and Indian Ocean outbreaks of 2005 and 2006, the global distribution of chikungunya virus (CHIKV) and the locations of epidemics have dramatically shifted. First, the Indian Ocean lineage (IOL) caused sustained epidemics in India and has radiated to many other countries. Second, the Asian lineage has caused frequent outbreaks in the Pacific islands and in 2013 was introduced into the Caribbean, followed by rapid spread to nearly all of the neotropics. Further, CHIKV epidemics, as well as exported cases, have been reported in central Africa after a long period of perceived silence. To understand these changes and to anticipate the future of the virus, the exact distribution, genetic diversity, transmission routes, and future epidemic potential of CHIKV require further assessment. To do so, we conducted the most comprehensive phylogenetic analysis to date, examined CHIKV evolution and transmission, and explored distinct genetic factors associated with the emergence of the East/Central/South African (ECSA) lineage, the IOL, and the Asian lineage. Our results reveal contrasting evolutionary patterns among the lineages, with growing genetic diversities observed in each, and suggest that CHIKV will continue to be a major public health threat with the potential for further emergence and spread. IMPORTANCE: Chikungunya fever is a reemerging infectious disease that is transmitted by Aedes mosquitoes and causes severe health and economic burdens in affected populations. Since the unprecedented Indian Ocean and Indian subcontinent outbreaks of 2005 and 2006, CHIKV has further expanded its geographic range, including to the Americas in 2013. Its evolution and transmission during and following these epidemics, as well as the recent evolution and spread of other lineages, require optimal assessment. Using newly obtained genome sequences, we provide a comprehensive update of the global distribution of CHIKV genetic diversity and analyze factors associated with recent outbreaks. These results provide a solid foundation for future evolutionary studies of CHIKV that can elucidate emergence mechanisms and also may help to predict future epidemics.

Chikungunya virus 3' untranslated region: adaptation to mosquitoes and a population bottleneck as major evolutionary forces.

The 3' untranslated genome region (UTR) of arthropod-borne viruses is characterized by enriched direct repeats (DRs) and stem-loop structures. Despite many years of theoretical and experimental study, on-going positive selection on the 3'UTR had never been observed in 'real-time,' and the role of the arbovirus 3'UTR remains poorly understood. We observed a lineage-specific 3'UTR sequence pattern in all available Asian lineage of the mosquito-borne alphavirus, chikungunya virus (CHIKV) (1958-2009), including complicated mutation and duplication patterns of the long DRs. Given that a longer genome is usually associated with less efficient replication, we hypothesized that the fixation of these genetic changes in the Asian lineage 3'UTR was due to their beneficial effects on adaptation to vectors or hosts. Using reverse genetic methods, we examined the functional importance of each direct repeat. Our results suggest that adaptation to mosquitoes, rather than to mammalian hosts, is a major evolutionary force on the CHIKV 3'UTR. Surprisingly, the Asian 3'UTR appeared to be inferior to its predicted ancestral sequence for replication in both mammals and mosquitoes, suggesting that its fixation in Asia was not a result of directional selection. Rather, it may have resulted from a population bottleneck during its introduction from Africa to Asia. We propose that this introduction of a 3'UTR with deletions led to genetic drift and compensatory mutations associated with the loss of structural/functional constraints, followed by two independent beneficial duplications and fixation due to positive selection. Our results provide further evidence that the limited epidemic potential of the Asian CHIKV strains resulted from founder effects that reduced its fitness for efficient transmission by mosquitoes there.

Urban epidemic of dengue virus serotype 3 infection, Senegal, 2009.

An urban epidemic of dengue in Senegal during 2009 affected 196 persons and included 5 cases of dengue hemorrhagic fever and 1 fatal case of dengue shock syndrome. Dengue virus serotype 3 was identified from all patients, and Aedes aegypti mosquitoes were identified as the primary vector of the virus.

Chikungunya virus emergence is constrained in Asia by lineage-specific adaptive landscapes.

Adaptation of RNA viruses to a new host or vector species often results in emergence of new viral lineages. However, lineage-specific restrictions on the adaptive processes remain largely unexplored. Recently, a Chikungunya virus (CHIKV) lineage of African origin emerged to cause major epidemics of severe, persistent, debilitating arthralgia in Africa and Asia. Surprisingly, this new lineage is actively replacing endemic strains in Southeast Asia that have been circulating there for 60 y. This replacement process is associated with adaptation of the invasive CHIKV strains to an atypical vector, the Aedes albopictus mosquito that is ubiquitously distributed in the region. Here we demonstrate that lineage-specific epistatic interactions between substitutions at amino acid positions 226 and 98 of the E1 envelope glycoprotein, the latter of which likely resulted from a founder effect, have for 60 y restricted the ability of endemic Asian CHIKV strains to adapt to this new vector. This adaptive constraint appears to be allowing invasion of the unoccupied vector niche by Ae. albopictus-adapted African strains. These results underscore how different adaptive landscapes occupied by closely related viral genotypes can profoundly affect the outcome of viral evolution and disease emergence.

Temporally structured metapopulation dynamics and persistence of influenza A H3N2 virus in humans.

Populations of seasonal influenza virus experience strong annual bottlenecks that pose a considerable extinction risk. It has been suggested that an influenza source population located in tropical Southeast or East Asia seeds annual temperate epidemics. Here we investigate the seasonal dynamics and migration patterns of influenza A H3N2 virus by analysis of virus samples obtained from 2003 to 2006 from Australia, Europe, Japan, New York, New Zealand, Southeast Asia, and newly sequenced viruses from Hong Kong. In contrast to annual temperate epidemics, relatively low levels of relative genetic diversity and no seasonal fluctuations characterized virus populations in tropical Southeast Asia and Hong Kong. Bayesian phylogeographic analysis using discrete temporal and spatial characters reveal high rates of viral migration between urban centers tested. Although the virus population that migrated between Southeast Asia and Hong Kong persisted through time, this was dependent on virus input from temperate regions and these tropical regions did not maintain a source for annual H3N2 influenza epidemics. We further show that multiple lineages may seed annual influenza epidemics, and that each region may function as a potential source population. We therefore propose that the global persistence of H3N2 influenza A virus is the result of a migrating metapopulation in which multiple different localities may seed seasonal epidemics in temperate regions in a given year. Such complex global migration dynamics may confound control efforts and contribute to the emergence and spread of antigenic variants and drug-resistant viruses.

A tissue-specific profile of miRNAs and their targets related to paeoniaflorin and monoterpenoids biosynthesis in Paeonia lactiflora Pall. by transcriptome, small RNAs and degradome sequencing.

Paeonia lactiflora Pall. (Paeonia) has aroused many concerns due to its extensive medicinal value, in which monoterpene glucoside paeoniflorin and its derivatives are the active chemical components. However, little is known in the molecular mechanism of monoterpenoids biosynthesis, and the regulation network between small RNAs and mRNAs in monoterpenoids biosynthesis has not been investigated yet. Herein, we attempted to reveal the tissue-specific regulation network of miRNAs and their targets related to paeoniaflorin and monoterpenoids biosynthesis in Paeonia by combining mRNA and miRNA expression data with degradome analysis. In all, 289 miRNAs and 30177 unigenes were identified, of which nine miRNAs from seven miRNA families including miR396, miR393, miR835, miR1144, miR3638, miR5794 and miR9555 were verified as monoterpenoids biosynthesis-related miRNAs by degradome sequencing. Moreover, the co-expression network analysis showed that four monoterpenoids-regulating TFs, namely AP2, MYBC1, SPL12 and TCP2, were putatively regulated by five miRNAs including miR172, miR828, miR858, miR156 and miR319, respectively. The present study will improve our knowledge of the molecular mechanisms of the paeoniaflorin and monoterpenoids biosynthesis mediated by miRNA to a new level, and provide a valuable resource for further study on Paeonia.

Transcriptomic and proteomic characteristics of the di(2-ethylhexyl) phthalate-induced sperm dna damage mouse model.

Di(2-ethylhexyl) phthalate (DEHP) is one of the most common organic pollutants and is added to various plastic products as a plasticizer. DEHP oxidative metabolite content in the human body is associated with DNA damage in sperm and decreased testosterone levels in blood. In this study, a DEHP-induced sperm DNA damage mouse model was replicated and improved, and the transcriptomic and proteomic characteristics of the model were observed. Male mice in the two groups were exposed to DEHP 1 g/kg/d or the same amount of normal saline for 60 days, and the sperm DNA fragmentation index (DFI) was detected by a sperm chromatin structure assay (SCSA). The mRNA and protein expression profiles of the testis were detected by RNA-seq and data-independent acquisition (DIA). The sperm DFI of the DEHP group was significantly increased. Compared with the control group, 111 differentially expressed genes (DEGs) and 2147 differentially expressed proteins (DEPs), such as Lamb2, Ahnak, Tkt, Dnah8 and Tbl2, were found in the DEHP group. These genes were mainly enriched in metabolic pathways, pathways in cancer and the PI3K-Akt signaling pathway. Our results showed that DEHP 1 g/kg/d can induce sperm DNA damage in a male mouse model after 60 days of intragastric administration. The reproductive toxicity of DEHP may be related to metabolic pathways in cancer and the PI3K-Akt signaling pathway.

Clinical study of Wuwei Fuzheng Yijing formula in the treatment of sperm DNA damage in male infertility: A study protocol for a randomized controlled trial.

BACKGROUND: The incidence of male infertility is increasing in recent years, and the semen routine examination of some patients is normal, but the semen pathological examination shows that the sperm DNA fragmentation index (DFI) is high, and the patients' clinical manifestations are infertility or recurrent abortion of their spouses. At present, there is no special treatment for male infertility caused by high DFI, and traditional Chinese medicine compound prescription shows potential value in the treatment of male infertility. Wuwei Fuzheng Yijing formula (WFY) is an effective prescription for the treatment of sperm DNA damage in male infertility, but there is no strict clinical trial to support its application. Therefore, we designed a randomized controlled trial to evaluate the efficacy and safety of WFY in patients with sperm DNA damage in male infertility. METHODS: In this randomized controlled study, 100 patients who met the inclusion criteria were randomly divided into WFY group and levocarnitine oral solution group. The treatment period was 12 weeks. The main observation index was sperm DFI, and the secondary observation index was sperm concentration, motility, survival rate, and TCM syndrome score. Safety observation indicators will include electrocardiogram, blood tests (including blood routine tests, liver and renal function), routine urine tests, and routine stool tests. All results were evaluated at the 4th and 8th week of the baseline, and the end of treatment. DISCUSSION: This study will provide a basis for the efficacy and safety of WFY in the treatment of sperm DNA damage in male infertility with spleen and kidney qi deficiency and blood stasis.

Genome-scale phylogenetic analyses of chikungunya virus reveal independent emergences of recent epidemics and various evolutionary rates.

Chikungunya virus (CHIKV), a mosquito-borne alphavirus, has traditionally circulated in Africa and Asia, causing human febrile illness accompanied by severe, chronic joint pain. In Africa, epidemic emergence of CHIKV involves the transition from an enzootic, sylvatic cycle involving arboreal mosquito vectors and nonhuman primates, into an urban cycle where peridomestic mosquitoes transmit among humans. In Asia, however, CHIKV appears to circulate only in the endemic, urban cycle. Recently, CHIKV emerged into the Indian Ocean and the Indian subcontinent to cause major epidemics. To examine patterns of CHIKV evolution and the origins of these outbreaks, as well as to examine whether evolutionary rates that vary between enzootic and epidemic transmission, we sequenced the genomes of 40 CHIKV strains and performed a phylogenetic analysis representing the most comprehensive study of its kind to date. We inferred that extant CHIKV strains evolved from an ancestor that existed within the last 500 years and that some geographic overlap exists between two main enzootic lineages previously thought to be geographically separated within Africa. We estimated that CHIKV was introduced from Africa into Asia 70 to 90 years ago. The recent Indian Ocean and Indian subcontinent epidemics appear to have emerged independently from the mainland of East Africa. This finding underscores the importance of surveillance to rapidly detect and control African outbreaks before exportation can occur. Significantly higher rates of nucleotide substitution appear to occur during urban than during enzootic transmission. These results suggest fundamental differences in transmission modes and/or dynamics in these two transmission cycles.