An increasing number of studies have reported that approximately 400 microRNAs (miRNAs), encoded by vertebrate viruses, regulate the expression of both host and viral genes. Many studies have used computational and/or experimental analyses to identify the target genes of miRNAs, thereby enabling us to understand miRNA functions. Here, we suggest that important aspects become apparent when we focus on conserved viral miRNAs, although these miRNA sequences generally show little similarity among viral species. Reliable viral miRNA–target gene pairs can be efficiently identified using evolutionary information. In this review, we summarize information on (i) the nucleotide sequence conservation among viral miRNAs and (ii) the RNAs targeted by viral miRNAs. Recent advances in these topics are discussed.

The aim of this study was to determine the usefulness of cancer antigen 125 (CA-125) serum levels in patients with tuberculosis (TB) with and without tuberculous serositis. A total of 64 TB patients with a mean age of 58.17 ± 19.05 years were enrolled in this observational case series study. All patients underwent blood sampling for the measurement of CA-125 serum levels before treatment. If the CA-125 serum levels were found to be elevated, the patients underwent blood sampling in the initial treatment phase, continuation treatment phase, and every 6 months thereafter for 2 years. The treatment outcomes of the pulmonary TB group were evaluated using chest radiography and sputum examinations, and those of the tuberculous serositis group were evaluated on the basis of the amounts of fluid determined by ultrasound. All patients in the tuberculous serositis group and 45% of the patients in the pulmonary TB group had elevated CA-125 serum levels before treatment. The pretreatment mean CA-125 serum level was significantly higher in the tuberculous serositis group than in the pulmonary TB group. CA-125 serum levels decreased along with improvement in anti-TB treatment outcomes in both the groups. In conclusion, the CA-125 serum levels in combination with clinical responses, chest radiography, and sputum examinations, can offer better monitoring of therapeutic responses in anti-TB treatment.

We report the results of the first large-scale questionnaire surveillance on the clinical use of pit viper antivenom in tertiary care centers in Japan. The questionnaire surveillance was conducted over a period of 3 years (April 2006 to March 2009). Completed questionnaires were received from the tertiary care centers of 108 (49.3%) medical institutions. In that period, 574 cases of pit viper bites, including 2 severe cases, were reported. Antivenom was administered in 44% of the cases of pit viper bites, and of these cases, 2.4% had adverse reactions but no severe symptoms. Approximately half of the clinicians indicated that antivenom was effective. Antivenom was recognized to be safe; however, the remarkable finding was that although the severity of treated cases was unclear, some clinicians reported using cepharanthine as the first choice of treatment for pit viper bites.

Since April 2009, the outbreak of human pandemic influenza A (H1N1) virus (pH1N1) infection has spread from North America to other parts of the world, and currently, pH1N1 is the predominant circulating strain of influenza viruses. Our objectives were to perform a serological survey of medical personnel at the Chumphae Hospital in Thailand and to investigate the prevalence of pH1N1 in randomly selected patients diagnosed with respiratory tract disease. Prevalence of pH1N1 in the patients was determined by performing real-time reverse transcription-polymerase chain reaction. The study was carried out between July 2009 and November 2010. Seroprevalence of hemaglutination inhibition (HI) titers among medical personnel was established in three cross-sectional studies at the end of each wave of the pandemic by performing HI assay to detect antibodies against pH1N1. Infection by the pH1N1 peaked between July and October 2009; the second wave was from January to March 2010 and the third wave from June to November 2010. The HI titers after the first, second, and third waves were 48.2%, 22.4%, and 25.7%, respectively. After the second and third waves, 52.1% and 45.3% of the medical personnel who had received pH1N1 vaccination had HI titers ≥40. These findings show that seasonal influenza strain in Chumphae and the predominant influenza strain from each wave was pH1N1. HI assay results also represent the severity of the attack rate in each wave.

Rabies is an endemic infectious disease and one of the most important causes of human mortality in both underdeveloped and developing countries. In Turkey, 167,000 individuals are believed to be victims of animal bites annually. In this study, we investigated Turkish physicians’ knowledge and clinical awareness of rabies caused by animal bites. This was a cross-sectional, analytical study. We used questionnaires that collected demographic information and assessed the physicians’ basic knowledge of rabies as well as the management of animal bites suspected of causing rabies. The questionnaires were completed in person with physicians who work in Istanbul. A total of 890 physicians responded to our cross-sectional questionnaires. The maximum possible scores for basic and clinical rabies- related knowledge was 100 points each. The average score for basic rabies knowledge was 64.5 ± 16, while the average score for clinical rabies knowledge was 62.8 ± 12. However, 68% of the physicians in the study were not aware of the proper method for cleaning wounds as a first-line treatment in postexposure prophylaxis. In addition, 38.4% of the physicians in the study did not understand the administration of vaccines together with immunoglobulin as part of postexposure prophylaxis. We also found that 79% of the physicians did not know the correct doses of vaccines, while 37.6% did not know the correct sites and routes of vaccine administration. Finally, 30% of the physicians were not aware of the correct vaccine schedules in postexposure prophylaxis. Our data indicate that Turkish physicians’ basic and clinical knowledge of rabies was insufficient. Rabies prophylaxis educational programs should be designed to educate physicians on the guidelines provided by the World Health Organization and the Advisory Committee on Immunization Practices for the treatment of rabies caused by animal bites.

This study was aimed at determining the molecular epidemiology of rabies virus (RABV) circulating in Vietnam. Intra vitam samples (saliva and cerebrospinal fluid) were collected from 31 patients who were believed to have rabies and were admitted to hospitals in northern provinces of Vietnam. Brain samples were collected from 176 sick or furious rabid dogs from all over the country. The human and canine samples were subjected to reverse transcription-polymerase chain reaction analysis. The findings showed that 23 patients tested positive for RABV. Interestingly, 5 rabies patients did not have any history of dog or cat bites, but they had an experience of butchering dogs or cats, or consuming their meat. RABV was also detected in 2 of the 100 sick dogs from slaughterhouses. Molecular epidemiological analysis of 27 RABV strains showed that these viruses could be classified into two groups. The RABVs classified into Group 1 were distributed throughout Vietnam and had sequence similarity with the strains from China, Thailand, Malaysia, and the Philippines. However, the RABVs classified into Group 2 were only found in the northern provinces of Vietnam and showed high sequence similarity with the strain from southern China. This finding suggested the recent influx of Group 2 RABVs between Vietnam and China across the border. Although the incidence of rabies due to circulating RABVs in slaughterhouses is less common than that due to dog bite, the national program for rabies control and prevention in Vietnam should include monitoring of the health of dogs meant for human consumption and vaccination for workers at dog slaughterhouses. Further, monitoring of and research on the circulating RABVs in dog markets may help to determine the cause of rabies and control the spread of rabies in slaughterhouses in Vietnam.

Yamakagashi, Rhabdophis tigrinus, is a natricine snake widely distributed in eastern Asia. Severe bite cases, some with fatal outcomes, occur regularly in Japan. Because previous production of R. tigrinus antivenom in rabbits and goats was quite effective, we considered the experimental manufacture of a new antivenom against R. tigrinus in horses. This new antivenom could be used in emergency treatment of snakebite victims. Two horses were immunized with venom extracted from about 500 snakes. After an adequate increase of the antivenom titer, serum was collected and subjected to standard purification procedures for the manufacture of equine antivenoms. The purified immunoglobulin fraction was freeze-dried in 1,369 vials under optimum conditions for therapeutic use. This antivenom proved to be very potent in neutralizing the coagulant and hemorrhagic activities of the snake venom. In cases of severe bites, this antivenom was used and recognized as effective even after the occurrence of severe symptoms.

Babesia microti, the primary causal agent of human babesiosis in North America, was thought to distribute in Europe in association with ixodid ticks and rodents. Recent analyses of β-tubulin and the eta subunit of the chaperonin-containing t-complex protein 1 (CCT7) genes revealed discrete clusters (a species-complex comprised of at least 4 taxa for the U.S., Kobe, Munich, and Hobetsu). To further assess the micro-evolutionary history and genetic variability within the taxon, we combined a set of 6 introns from the CCT7 gene to use as a rapidly evolving DNA marker. Phylogenetic and comparative sequence analyses subdivided the U.S. taxon into 3 geographic subclades―North America, western to central Eurasia, and northeastern Eurasia (≥98% bootstrap supports for each node). The Kobe taxon, which occurs only in a few geographic foci of Japan, could further be subdivided into 2 subgroups (100% support). The Munich and Hobetsu taxa, common to Europe and Japan, respectively, exhibited little or no pairwise sequence divergence among geographically diverse samples, suggesting an extreme population bottleneck during recent history. Despite the small sample size, this study provides a better understanding of the micro-evolutionary relationships and the genetic variability present within each lineage of the B. microti-group.

The aim of this study was to evaluate the prevalence and risk factors of human papillomavirus (HPV) infection among human immunodeficiency virus (HIV)-positive women in China. To this end, we enrolled 200 HIV-positive and 182 HIV-negative women in this cross-sectional cohort study. The following sampling methods were used: (i) structured interview, (ii) CD4 cell counts, and (iii) cervical specimens. HPV genotype (total 23 types) was analyzed using polymerase chain reaction assay. Logistic regression analysis was used to identify independent causative factors for HPV infection. The prevalence of HPV infection was 3-fold higher in the HIV-positive women than in the HIV-negative women. The overall prevalences of HPV infection, high risk (HR)-HPV infection, and multiple HPV infections in the HIV-positive women were 36.5%, 33.5%, and 13.0%, respectively, and the corresponding values in HIV-negative women were 12.1%, 10.4%, and 6.0%, respectively (P < 0.05). The types of HR-HPVs were similar in the HIV-positive and HIV-negative women (HPV-16, -52, -58, and -18), and the prevalences of infections by these viruses were 1.5- to 3-fold higher in the HIV-positive group than in the HIV-negative group. HR-HPV infection among the HIV-positive women was associated with three factors: low CD4 count (OR for 200 ≤ CD4 ≤ 350 and CD4 < 200/µL were 2.11 and 3.13, respectively), HIV infection through sexual contact (OR, 7.90; 95% CI, 2.38–14.60), and having HIV-positive sexual partners (OR, 2.02; 95% CI, 1.03–3.95). We found that the prevalence of HPV infection among the HIV-positive Chinese women was higher than that among the HIV-negative women; moreover, among the HIV-positive women, factors associated with HIV infection were risk factors for HR-HPV infection.

This study aimed to investigate the epidemiology of norovirus (NoV) associated-diarrhea among pediatric outpatients in Shanghai, and characterize the genotypes of circulating NoV strain. Stool samples were collected from 910 children with non-dysenteric diarrhea between August 2008 and July 2009. One-step real-time RT-PCR was used to screen for NoV genogroup I (GI) and genogroup II (GII). Genotypes were classified by sequence analysis of partial capsid and RNA-dependent RNA polymerase (RdRp) fragments. NoV was detected year round with high activity in July, August, September, and October. Of 910 specimens, 165 (18.13%) were positive for NoV; 4 (2.42%) were GI, and 161 (97.58%) were GII. Based on capsid sequences, 8 different genotypes were identified for 114 NoV strains, including GII.4 2006b (57.89%), GII.3 (30.70%), GII.6 (4.39%), GII.12 (3.51%), GII.14, GII.2, GI.4, and GI.5 (0.88% each). Based on the RdRp sequences of 86 NoV strains, NoV was genotyped as GII.4 (62.79%), GII.12 (30.23%), GII.g (2.33%), GII.2 (1.16%), GII.6 (1.16%), GII.7 (1.16%), and GI.4 (1.16%). The RdRp genotypes of 30 strains were inconsistent with the capsid genotypes, indicating potential NoV recombinants.

Twenty-three Chinese men with primary HIV infection (PHI) who have sex with men, and 17 asymptomatic HIV-infected patients participated in this study. The effect of different initial activation parameters on viral set points and the immunologic progression of subjects with PHI were determined. We found that the proportions of CD38+DR+ and CD38−DR+ T cells were lower while those of CD38+DR− and CD38−DR− T cells were higher in patients with PHI than in the asymptomatic HIV-infected patients. The initial proportion of CD4+CD38+DR+ T cells emerged as a statistically significant predictor (P < 0.05) of viral set points by multivariate least squares regression. In addition, individuals with a higher initial proportion of CD4+CD38+DR+ cells were more likely to have CD4+ T cell counts decrease to ≤350 cells/µl (P = 0.015), as determined by Kaplan-Meier curve analysis. We concluded that the initial CD4+CD38+DR+ T cell level in PHI patients is an important prognostic parameter of viral set points and is therefore related to disease progression.

Nontuberculous mycobacteria are known to produce soft-tissue infections following surgical procedures. We report a non-healing surgical wound infection caused by a scotochromogenic nontuberculous mycobacterium, possibly Mycobacterium flavescens, which was earlier thought to be saprophytic. This organism was isolated from the wound discharge at the site of incision following a laparoscopic cholecystectomy; the finding suggests that M. flavescens is a clinically important pathogen in cases of surgical wound infections.

We report the case of a 62-year-old man who developed eosinophilic pneumonia due to visceral larva migrans (VLM) that was possibly caused by Ascaris suum. The patient, a resident of the middle Kyushu area who was found of eating raw porcine liver, complained of dry cough without dyspnea. The chest radiography showed a migration of infiltrative shadow. Transbronchial lung biopsy of the right middle lobe revealed massive infiltration of eosinophils. The multi-dot enzyme-linked immunosorbent assay (ELISA) and microtiter plate ELISA showed positive results for A. suum; therefore, the patient was diagnosed with VLM caused by A. suum. The patient was administered albendazole (600 mg/day) for 28 days; he recovered successfully with no adverse effects except mild liver dysfunction. Several cases of VLM caused by A. suum have been reported in Japan, with a majority of the cases being reported in Kyushu. Careful history taking of the patient's area of residence and dietary habit is essential for the diagnosis of this parasitic disease with underestimated prevalence.

Several studies in Western countries have found that lower respiratory tract infections (LRTIs) caused by the respiratory syncytial virus (RSV) in infancy may subsequently trigger the development of asthma. In this study, we enrolled 262 infants under the age of 3 who had been admitted to our hospital with LRTI between September 2002 and August 2003. RSV infection was diagnosed in these patients using an RSV rapid diagnostic kit and by measuring antibody titers in paired serum samples. In March 2009, we sent questionnaires on post-discharge allergic conditions to the families of the 249 patients, excluding 13 who had a prior history of asthma. A total of 133 responses were received (response rate, 53.4%); RSV was detected in 36 patients of the RSV group and 97 patients of the non-RSV group. Wheezing was experienced post-discharge by 10 patients in the RSV group (27.8%) and 32 in the non-RSV group (33.0%) (P = 0.57). Four patients in the RSV group (11.1%) and 6 in the non-RSV group (6.2%) (P = 0.34) were treated for asthma. This study revealed that RSV LRTI in infancy does not predispose children to subsequent development of asthma at the age of 7 years and 7 months. We believe that this is the first Japanese survey that has examined the relationship between RSV LRTI in infancy and the subsequent development of asthma.

Since 2002, we have conducted a series of intervention programs in Sado Island, Japan, to increase awareness of the appropriate use of antimicrobials among public and health-care staff. The objective of our study was to determine the antimicrobial susceptibility of Staphylococcus aureus among children in a semi-closed geographic region with controlled antimicrobial use. The average hospital visit-based rate for antimicrobial drug prescription drastically decreased from 535 per 1,000 clinic visits before the intervention programs to 45–75 per 1,000 clinic visits after the intervention programs (P < 0.0001). We obtained 1,260 clinical isolates of S. aureus from 957 pediatric outpatients aged 0–15 years between 2002 and 2010 at Sado General Hospital. The prevalence of methicillin-resistant S. aureus (MRSA) in the final year of the study (2010, 7.9%) was significantly lower than that in the early period of the study (2003, 23.6%; P < 0.001). All tested S. aureus isolates were susceptible to trimethoprim/sulfamethoxazole, teicoplanin, linezolid, and vancomycin. The prevalence of gentamicin-resistant S. aureus in isolates from skin lesions was significantly higher than in isolates from other lesions (41.3% versus 28.5%; P < 0.05). In conclusion, prevalence of MRSA in children in our study population could be decreased by eliminating unnecessary antimicrobial drug prescriptions.

Crimean-Congo hemorrhagic fever virus (CCHFV), a member of the genus Nairovirus of the family Bunyaviridae, causes a severe disease in humans with high mortality rates. In Turkey, the number of patients with CCHF has increased since 2002. Here, we aimed to treat CCHF patients with CCHFV hyperimmunoglobulin. We prepared a CCHFV hyperimmunoglobulin product from 22 individuals who survived CCHF infection. A total of 26 CCHF patients were enrolled into this study. For CCHFV hyperimmunoglobulin administration, a Kubar Unit (KU) was defined. As a standard therapeutic approach, 400 KU of hyperimmunoglobulin were given to each patient as a single dose before viral load was detected. We used one-step real-time reverse transcriptase-PCR to monitor the viral load of CCHF patients. According to the one-step real-time PCR results, 15 patients with a viral load of 10⁸ copies/mL or more were defined as high risk. In this high-risk group, the survival rate was found to be 86.6% (13/15) and 2 patients died despite CCHFV hyperimmunoglobulin administration. CCHF is a very serious and highly fatal infection, particularly for patients in the defined high-risk group. Prompt administration of CCHFV hyperimmunoglobulin might be a very promising new treatment approach, especially for high-risk individuals.