Characterization of In vitro inhibitory effects of consensus short interference RNAs against non-structural 5B gene of hepatitis C virus 1a genotype

Purpose: Chronic hepatitis C has infected approximately 170 million people worldwide. The novel direct-acting antivirals have proven their clinical efficacy to treat hepatitis C infection but still very expensive and beyond the financial range of most infected patients in low income and even resource replete nations. This study was conducted to establish an in vitro stable human hepatoma 7 (Huh-7) cell culture system with consistent expression of the non-structural 5B (NS5B) protein of hepatitis C virus (HCV) 1a genotype and to explore inhibitory effects of sequence-specific short interference RNA (siRNA) targeting NS5B in stable cell clones, and against viral replication in serum-inoculated Huh-7 cells. Materials and Methods: In vitro stable Huh-7 cells with persistent expression of NS5B protein was produced under gentamycin (G418) selection. siRNAs inhibitory effects were determined by analysing NS5B expression at mRNA and protein level through reverse transcription-polymerase chain reaction (PCR), quantitative real-time PCR, and Western blot, respectively. Statistical significance of data (NS5B gene suppression) was performed using SPSS software (version 16.0, SPSS Inc.). Results: siRNAs directed against NS5B gene significantly decreased NS5B expression at mRNA and protein levels in stable Huh-7 cells, and a vivid decrease in viral replication was also exhibited in serum-infected Huh-7 cells. Conclusions: Stable Huh-7 cells persistently expressing NS5B protein should be helpful for molecular pathogenesis of HCV infection and development of anti-HCV drug screening assays. The siRNA was effective against NS5B and could be considered as an adjuvant therapy along with other promising anti-HCV regimens.

Role of biopsy in pediatric lymphadenopathy

To determine the role of lymph node biopsy in the diagnosis of lymphadenopathy and to find out the pattern of different diseases in relation to age, gender, and the site of lymph nodes involved. This retrospective study was carried out at the Histopathology Department of the Children's Hospital and The Institute of Child Health, Lahore, Pakistan, over a period of 9 years, from January 1999 to December 2007. Tissue samples were collected from 898 children presenting with lymphadenopathy, and the diagnosis was confirmed on histology and through various specific tests. The clinical data of the patients were collected from computerized hospital records. Among the total 898 consecutive lymph node biopsies, the most common pathology encountered was reactive hyperplasia in 356 children [39.6%], followed by tuberculosis in 262 [29.1%] and malignant lymphomas in 132 children [14.6%]. The rest of the lesions include; 72 cases of granulomatous lymphadenitis [8%], 13 of histiocytosis X [1.4%], 44 [4.9%] of metastatic tumors, 16 of chronic inflammation [1.8%], and 3 cases of Kikuchi's disease [0.3%]. The cause of lymphadenopathy was found to be significantly associated with age, gender, and site of the lymph nodes involved. Lymphadenopathy is a relatively common condition in the pediatric age group. Although 39.6% of children had reactive hyperplasia of unknown etiology, 60.3% children presented with a specific diagnosis

Smith - Lemli - Opitz Syndrome

This is a very rare autosomal recessive disorder characterised by short stature, microcephaly,low set ears, hypospadias (in males), pyramidal signs in addition to several other features as described.

Incidence of intestinal protozoan parasites of man in twin cities of Rawalpindi Islamabad

A total of 8850 stool samples [Rawalpindi = 5360 and Islamabad = 3490] from seven hospitals of the twin cities of Rawalpindi-Islamabad were examined for intestinal prtotozoa during September 1980 to September 1981 and the following protozoan parasites were recorded: Entamoeba histolytica, E. coli, iodamoeba buetschlli, Giardia lamblia, and Pentarichomonas hominis. Of the total patients examined 41.88% from Rawalpindi and 42.14% from Islamabad carried intestinal protozoan infection. In both the cities E. histolytica and G. lamblia had an almost similar incidence while E. coli, I. Buetschlii, and P. hominis showed a very low percentage of infection. It was noted that such factors as heavy faecal pollution of the premises, unsanitary faecal disposal, unhygienic water supply, and poor personal cleanliness facilitate transmission of these parasites