Resumo O vírus da hepatite C (HCV) é o grave problema de saúde pública das doenças hepáticas. Aproximadamente 170 milhões de pessoas no mundo estão infectadas com HCV; no Paquistão, a doença tem alto índice de ocorrência. O presente estudo prevê uma prevalência atualizada do HCV e distribuição genotípica na população geral do distrito de Mardan, Khyber Pakhtunkhwa (KP), Paquistão. As amostras de sangue de 6.538 indivíduos, incluindo 3.263 homens e 3.275 mulheres, foram analisadas para o antígeno de superfície da hepatite C por teste imunocromatográfico (ICT), ensaio imunoenzimático (ELISA) e reação em cadeia da polimerase de transcrição reversa (PCR). Verificou-se que 396 (12,13%) de 3.263 indivíduos continham anticorpos no sangue contra o HCV, enquanto entre as diferentes faixas etárias as maiores incidências de anticorpos anti-HCV foram encontradas na faixa etária de 31 a 40 anos (11,01%). As amostras positivas para ICT foram posteriormente rastreadas por nested PCR para determinar a existência de HCV-RNA ativo. Identificou-se que 7,11% (3.263) do total da população (6.538) testada foram positivos, dentre os quais 461 (14,07%) mulheres possuíam anticorpos no sangue contra o HCV. Nossos dados mostraram que a infecção total pelo HCV na população investigada foi de 5,78%. Maior porcentagem de prevalência de HCV foi detectada em homens do que em mulheres nas faixas etárias de 31-40 e 41-50. Para comparar a prevalência de genótipos de HCV com relação à idade no genótipo masculino e feminino 3a foi encontrado o genótipo mais prevalente seguido por 1a, 2a e 3b, respectivamente.

Abstract Hepatitis C virus (HCV) is the serious global public health burden of liver disease. Approximately 170 million people in the world are infected with (HCV). In Pakistan, where the disease has high occurrence rate. The present study envisages an up-to-date prevalence of HCV and genotypic distribution in the general population of Mardan District, Khyber Pakhtunkhwa (KP), Pakistan. The blood samples from 6,538 individuals including 3,263 males and 3,275 females were analyzed for hepatitis C surface antigen by Immuno-chromatographic test (ICT), Enzyme-linked immunosorbent assay (ELISA), and reverse transcription-polymerase chain reaction (PCR). It was found that 396 (12.13%) out of 3263 individuals contained antibodies in their blood against HCV, while among the different age groups, the highest incidences of HCV antibodies were found in the 31-40 age group (11.01%). The ICT positive samples were further screened by nested PCR to determine the existence of active HCV-RNA. It was identified that 7.11% (3263) of the total population (6538) tested was positive, among which the 461 (14.07%) females possessed antibodies in their blood against HCV. Our data showed total HCV infection in the investigated population was 5.78%. Higher percentage of HCV prevalence was detected in males than females in the age group 31-40 and 41-50. To compare the prevalence of HCV genotypes age-wise in male and female genotype 3a was found most prevalent genotype followed by 1a, 2a and 3b, respectively.

Resumo A infecção pelo vírus da hepatite B é perigosa entre os cinco tipos de hepatite, pois permanece clinicamente assintomática. O presente estudo traça a prevalência atualizada do vírus da hepatite B (HBV) na população geral de Mardan, Khyber Pakhtunkhwa, no Paquistão. Amostras de sangue de 4.803 indivíduos, incluindo 2.399 homens e 2.404 mulheres, foram investigadas. Todas as amostras suspeitas foram analisadas para o antígeno de superfície da hepatite B usando teste imunocromatográfico (ICT), enzyme-linked immunosorbent assay (ELISA), seguido por transcrição reversa-reação em cadeia da polimerase (RT-PCR). Os resultados mostraram que 312 (13,00%) de 2.399 indivíduos continham anticorpos no sangue contra o VHB, enquanto, entre as diferentes faixas etárias, as maiores incidências de anticorpos VHB foram encontradas nos grupos de 21 a 30 anos (10,73%). Além disso, amostras positivas para ICT foram rastreadas por reação em cadeia da polimerase aninhada para detectar a existência de HBV-DNA ativo. Observou-se que 169 (7,04%) de 2.399 homens do total da população (4803) testados foram positivos. Por outro lado, 463 mulheres (19,25%) possuíam anticorpos no sangue contra VHB. Acumulativamente, nossos resultados mostraram uma porcentagem maior de prevalência de HBV em homens do que em mulheres na faixa etária de 21 a 30 anos. O total de HCV infectados na população geral de Mardan foi registrado em 5,7%, incluindo homens e mulheres.

Abstract Hepatitis B virus infection is perilous among the five types of Hepatitis, as it remains clinically asymptomatic. The present study draws up-to-date prevalence of Hepatitis B virus (HBV) in the general population of Mardan, Khyber Pakhtunkhwa Pakistan. The blood samples from 4803 individuals including 2399 male and 2404 females were investigated. All the suspected samples were analyzed for hepatitis B surface antigen using Immuno-chromatographic test (ICT), Enzyme-linked immunosorbent assay (ELISA), and followed by Reverse transcription-polymerase chain reaction (RT-PCR). Results showed that 312 (13.00%) out of 2399 individuals contained antibodies in their blood against HBV, while among the different age groups, the highest incidences of HBV antibodies were found in the age of 21-30 groups (10.73%). Furthermore, the ICT positive samples were screened by nested polymerase chain reaction to detect the existence of active HBV-DNA. It was observed that 169 (7.04%) out of (2399) male of the total population (4803) tested was positive. On the other hand, the female 463 (19.25%) possessed antibodies in their blood against HBV. Accumulatively, our results showed a higher percentage of HBV prevalence in males than females in the age group 21-30 years. The total HCV infected in Mardan general population was recorded at 5.7% comprising both male and female.

RESUMO O objetivo do presente estudo foi investigar a influência de mudanças patológicas de plasma antes de inseminação artificial (TAI) na gestação de vacas. O conteúdo de estrogênio (E2), progesterona (P4), glucose (Glu), selênio (Se), fator neurotrófico derivado do cérebro (BDNF), e histamina (HIS) no plasma de 48 vacas Holstein foi medido antes de TAI. De acordo com a detecção de estro, as vacas foram divididas em dois grupos: estro (E) e anestro (A). Após teste de gestação 28 d após TAI, dois grupos de E e A foram formados em gestação positiva do grupo E (EP+), gestação negativa do grupo E (EP-), gestação positiva do grupo A (AP+), e gestação negativa do grupo A (AP-). Os valores de E2, P4, Glu, Se, BDNF e hIS foram significativamente diferentes entre os quatro grupos (P<0,01). A análise ROC foi utilizada para determinar o risco de teste de gestação negativo (-) após aumento de TAI quando plasma E2 estava abaixo de 46,45 pmol/L em vacas antes de TAI. Alterações em E2, P4,hIS, Glu e BDNF no sangue de estro natural e anestro natural em vacas afetou a gestação após TAI. O nível de E2 no plasma pode ser usado para avaliar o risco de gestação negativa após TAI.

ABSTRACT The objective of this study was to investigate the influence of plasma pathological changes before timed artificial insemination (TAI) on pregnancy of cows. The contents of estrogen (E2), progesterone (P4), glucose (Glu), selenium (Se), brain-derived neurotrophic factor (BDNF), and histamine (HIS) in plasma of 48 Holstein cows were measured before TAI. According to the estrus detection, the cows were divided into estrus (E) and anestrus (A) groups. After pregnancy testing at 28 d after TAI, two groups of E and A were divided into positive pregnancy of E group (EP+), negative pregnancy of E group (EP-), positive pregnancy of A group (AP+), and negative pregnancy of A group (AP-). The contents of E2, P4, Glu, Se, BDNF and hIS significantly differed among the four groups (P<0.01). The ROC analysis was used to determine the risk of negative pregnancy test (-) after TAI was increased when plasma E2 was less than 46.45 pmol/L in cows before TAI. The changes in E2, P4,hIS, Glu, and BDNF in the blood of natural estrus and natural anestrus cows affected the pregnancy after TAI. the level of E2 in plasma may be used to assess the risk of negative pregnancy after TAI.

Abstract Background: Cyclocodon lancifolius (Roxb.) Kurz is a perennial medicinal and edible plant with a huge potential economic value. The wild resources of this plant are gradually scarce by the serious destruction of the habitat and the limitations of sexual reproduction. This is the first attempt to establish an in vitro reproductive system for the species. Hypothesis: The suitable plant regulator types and its mass concentration range, combined with the explants, can induce the development of plants at various stages. We expected to establish an in vitro regeneration system of C. lancifolius based on these factors. Species studied: Cyclocodon lancifolius Study site and years of study: Yunnan Breeding and Cultivation Research and Development Center of Endangered and Daodi Chinese Medicinal Materials, Yunnan University of Chinese Medicine, from 2017 to 2019. Methods: The plant regeneration of C. lancifolius was established by single factor, L9 (34) orthogonal and complete combination experiments. Results: Stem segments were the best explants for callus induction on MS medium containing 0.05 mg·L-1 KT, 0.5 mg·L-1 6-BA and 0.5 mg·L-1 NAA. MS medium with 0.2 mg·L-1 TDZ and MS medium were used alternately as the culture method to conduct differentiation and proliferation of adventitious shoot. The optimal protocol for the rooting was MS medium combined with 0.1 mg·L-1 6-BA and 1.0 mg·L-1 NAA. Conclusions: A rapid propagation system of C. lancifolius was established which provided a possible solution for the protection of wild resources and artificial cultivation.

Resumen Antecedentes: Cyclocodon lancifolius es una planta perenne medicinal y comestible. Los recursos silvestres de esta planta escasean gradualmente debido a la grave destrucción del hábitat y las limitaciones de la reproducción. Este es el primer intento de establecer un sistema reproductivo in vitro para esta especie. Hipótesis: El tipo de regulador de crecimiento adecuado y su rango de concentración másica, combinado con los explantes, pueden inducir el crecimiento de plantas en varias etapas. Se espera establecer un sistema de regeneración in vitro de C. lancifolius basado en estos factores. Especie estudiadas: Cyclocodon lancifolius. Sitio y años del estudio: Universidad de Medicina China de Yunnan, De 2017 a 2019. Métodos: La regeneración de plantas de C. lancifolius se estableció mediante los experimentos de un solo factor, ortogonales de tipo L9 (34) y los de combinación completa. Resultados: Los segmentos del tallo fueron los mejores explantes para la inducción de callos en un medio MS que contenía 0.05 mg·L-1 KT, 0.5 mg·L-1 6-BA y 0.5 mg·L-1 NAA. El medio MS + 0.2 mg·L-1 TDZ se usó de manera alternativa con el medio MS como el método de cultivo para realizar la diferenciación y proliferación de brotes adventicios. El protocolo óptimo para el enraizamiento fue el medio MS combinado con 0.1 mg·L-1 6-BA y 1.0 mg·L-1 NAA. Conclusiones: Se estableció un sistema de propagación rápida de C. Lancifolius que proporcionó una posible solución para la protección de los recursos silvestres, así como para el cultivo artificial.

Background: Cold-active endo-1, 4-β-glucanase (EglC) can decrease energy costs and prevent product denaturation in biotechnological processes. However, the nature EglC from C. farmeri A1 showed very low activity (800 U/L). In an attempt to increase its expression level, C. farmeri EglC was expressed in Escherichia coli as an N-terminal fusion to protein S (ProS) from Myxococcus xanthus. Results: A novel expression vector, pET(ProS-EglC), was successfully constructed for the expression of C. farmeri EglC in E. coli. SDS-PAGE showed that the recombinant protein (ProS-EglC) was approximately 60 kDa. The activity of ProS-EglC was 12,400 U/L, which was considerably higher than that of the nature EglC (800 U/L). ProS-EglC was active at pH 6.5-pH 8.0, with optimum activity at pH 7.0. The recombinant protein was stable at pH 3.5-pH 6.5 for 30 min. The optimal temperature for activity of ProS-EglC was 30°C-40°C. It showed greater than 50% of maximum activity even at 5°C, indicating that the ProS-EglC is a cold-active enzyme. Its activity was increased by Co2+ and Fe2+, but decreased by Cd2+, Zn2+, Li+, methanol, Triton-X-100, acetonitrile, Tween 80, and SDS. Conclusions: The ProS-EglC is promising in application of various biotechnological processes because of its cold-active characterizations. This study also suggests a useful strategy for the expression of foreign proteins in E. coli using a ProS tag.

Agaricus blazei Murill is a native Brazilian mushroom which functions primarily as an anticancer substance in transplanted mouse tumors. However, the mechanism underlying this function of A. blazei Murill remains obscure. The present study was carried out to investigate the effect of fraction FA-2-b-ß, an RNA-protein complex isolated from A. blazei Murill, on human leukemia HL-60 cells in vitro. Typical apoptotic characteristics were determined by morphological methods using DNA agarose gel electrophoresis and flow cytometry. The growth suppressive effect of fraction FA-2-b-ß on HL-60 cells in vitro occurred in a dose- (5-80 µg/mL) and time-dependent (24-96 h) manner. The proliferation of HL-60 cells (1 x 10(5) cells/mL) treated with 40 µg/mL of fraction FA-2-b-ß for 24-96 h and with 5-80 µg/mL for 96 h resulted in inhibitory rates ranging from 8 to 54.5%, and from 4.9 to 86.3%, respectively. Both telomerase activity determined by TRAP-ELISA and mRNA expression of the caspase-3 gene detected by RT-PCR were increased in HL-60 cells during fraction FA-2-b-ß treatment. The rate of apoptosis correlated negatively with the decrease of telomerase activity (r = 0.926, P < 0.05), but correlated positively with caspase-3 mRNA expression (r = 0.926, P < 0.05). These data show that fraction FA-2-b-ß can induce HL-60 cell apoptosis and that the combined effect of down-regulation of telomerase activity and up-regulation of mRNA expression of the caspase-3 gene could be the primary mechanism of induction of apoptosis. These findings provide strong evidence that fraction FA-2-b-ß could be of interest for the clinical treatment of acute leukemia.

OBJECTIVE: To assist with strategic planning for the eradication of malaria in Henan Province, China, which reached the consolidation phase of malaria control in 1992, when only 318 malaria cases were reported. METHODS: We conducted a prospective two-year study of the costs for Henan's malaria control programme. We used a cost model that could also be applied to other malaria programmes in mainland China, and analysed the cost of the three components of Henan's malaria programme: suspected malaria case management, vector surveillance, and population blood surveys. Primary cost data were collected from the government, and data on suspected malaria patients were collected in two malaria counties (population 2 093 100). We enlisted the help of 260 village doctors in six townships or former communes (population 247 762), and studied all 12 325 reported cases of suspected malaria in their catchment areas in 1994 and 1995. FINDINGS: The average annual government investment in malaria control was estimated to be US$ 111 516 (case-management 59%; active blood surveys 25%; vector surveillance 12%; and contingencies and special projects 4%). The average cost (direct and indirect) for patients seeking treatment for suspected malaria was US$ 3.48, equivalent to 10 days' income for rural residents. Each suspected malaria case cost the government an average of US$ 0.78. CONCLUSION: Further cuts in government funding will increase future costs when epidemic malaria returns; investment in malaria control should therefore continue at least at current levels of US$ 0.03 per person at risk.

OBJETIVO: Contribuir mediante la planificación estratégica a la erradicación del paludismo en la provincia china de Henan, donde se alcanzó la fase de consolidación del control del paludismo en 1992, año en que sólo se notificaron 318 casos de la enfermedad. MÉTODOS: Realizamos un estudio prospectivo de dos años sobre los costos del programa de lucha antipalúdica de Henan. Para ello utilizamos un modelo de costos que pudiera aplicarse también a otros programas contra el paludismo en la China continental, y analizamos el costo de los tres componentes del programa: manejo de los casos presuntos de paludismo, vigilancia de los vectores, y encuestas hematológicas de la población. Los datos principales sobre los costos se obtuvieron de la Administración, y los datos sobre los pacientes presuntamente afectados proceden de dos distritos con paludismo (2 093 100 habitantes). Conseguimos que nos ayudaran 260 médicos rurales en seis municipios o antiguas comunas (247 762 habitantes), y estudiamos la totalidad de los 12 325 casos notificados de posible paludismo registrados en sus zonas de captación en 1994 y 1995. RESULTADOS: Se calculó que la inversión anual media del Gobierno en el control del paludismo ascendía a US$ 111 516 (manejo de casos, 59%; encuestas hematológicas activas, 25%; vigilancia de vectores, 12%; e imprevistos y proyectos especiales, 4%). El gasto medio (directo e indirecto) para los pacientes que solicitaron tratamiento sospechando que tenían paludismo fue de US$ 3,48, suma que para los habitantes de las zonas rurales equivalía a los ingresos de 10 días. Cada caso presunto de paludismo supone para la Administración un gasto medio de US$ 0,78. CONCLUSIÓN: Todo recorte adicional de los fondos públicos se traducirá en mayores costos en el futuro cuando retorne el paludismo epidémico; por consiguiente, la inversión en la lucha antipalúdica debe mantenerse como mínimo en los niveles actuales de US$ 0,03 por persona en riesgo.

OBJECTIF: Aider à la planification stratégique de l'éradication du paludisme dans la Province de Henan (Chine), où a été atteinte la phase de consolidation de la lutte antipaludique en 1992, date à laquelle seuls 318 cas de paludisme ont été notifiés. MÉTHODES: Nous avons réalisé une étude prospective de deux ans sur les coûts du programme de lutte antipaludique de la Province de Henan. Nous avons utilisé pour cela un modèle de coût qui peut également être appliqué à d'autres programmes de lutte antipaludique en Chine continentale, et avons analysé le coût des trois éléments du programme, à savoir la prise en charge des cas suspects de paludisme, la surveillance des vecteurs et les enquêtes hématologiques en population. Les données primaires sur les coûts ont été obtenues auprès du Gouvernement, et les données sur les cas suspects ont été recueillies dans deux districts touchés par le paludisme (2 093 100 habitants). Nous avons fait appel à 260 médecins de village dans six municipalités ou ex- communes (247 762 habitants) et étudié la totalité des 12 325 cas suspects de paludisme notifiés dans la zone desservie par ces médecins en 1994 et 1995. RÉSULTATS: L'investissement annuel moyen du Gouvernement dans la lutte antipaludique a été évalué à US$ 111 516 (prise en charge des cas 59%; enquêtes hématologiques 25%; surveillance des vecteurs 12%; imprévus et projets spéciaux 4%). Le coût moyen (direct et indirect) pour les patients se faisant soigner pour une suspicion de paludisme était de US$ 3,48, soit l'équivalent de 10jours de revenu pour les résidents des zones rurales. Chaque cas suspect de paludisme coûtait en moyenne au Gouvernement US$ 0,78. CONCLUSION: Toute nouvelle réduction des fonds publics augmentera le coût futur de la lutte en cas de retour d'épidémies de paludisme; l'investissement dans la lutte antipaludique doit donc être maintenu au moins au niveau actuel de US$ 0,03 par personne à risque.