Synthesis and anti-HCC activity of full 2ʹ-F/OMe-siRNA encapsulated with neutral cytidinyl/cationic lipid / 药学学报

A variety of full 2ʹ-F/OMe-modified siRNAs were designed and synthesized, and the activity against hepatocellular carcinoma Huh-7 and HepG2 cells was evaluated. K&A DNA/RNA H-8 synthesizer was used to synthesize siRNAs, and neutral cytidinyl lipid DNCA mixed with cationic lipid CLD were used to transfect siRNA. By RT-qPCR and CCK-8 assay, the target gene silence and the proliferation of Huh-7 and HepG2 cells were detected. The siRNAs loading into Ago2 protein was detected by RNA-binding protein immunoprecipitation. Drug uptake and cell apoptosis were detected by flow cytometry, and the expression of PLK1 protein was detected by Western blot. Partial full 2ʹ-F/OMe modified siRNAs, especial siPLK1A3, increased the uptake of Huh-7 cells, enhanced their binding to Ago2 and gene silencing activity, down-regulated PLK1 protein, as well as induced more Huh-7 cell apoptosis and proliferation inhibition activity. It provides important data for the development of novel siRNA modification patterns and anti-HCC formulations.

A Study On Etiology And Clinical Profile Of Patients With Thrombocytopenia

BACKGROUND: Thrombocytopenia refers to a reduction in platelet count below 1.5 lakh/microliter. The presence of thrombocytopenia in a hemogram should alert the physician to identify the underlying etiology for the prompt management of the patient. Timely identification and treatment prevent bleeding manifestations, requirement of platelet transfusions/steroids and overall impact on mortality of the patients. AIM OFSTUDY:Analysis to study the etiology, bleeding manifestation, percentage of patients requiring platelet transfusion, length of hospital stay in patients with thrombocytopenia. METHODOLOGY: 100 cases thrombocytopenia both male and female were included in the study. The diagnosis was made on peripheral smear and Hemogram. RESULTS: Dengue fever was the most common cause of thrombocytopenia with 43 cases. Sepsis with 23 cases was the second commonest. Bleeding manifestations were seen in 23% of the study population.100% of the patients with platelet count less than 10,000/microlitre had bleeding manifestations. 26 patients (26%) received platelet transfusion out of which 23 were therapeutic and 3 were prophylactic transfusions. Steroid therapy was given in 11% of patients. Mortality was highest in patients with sepsis induced thrombocytopenia. CONCLUSION:This study shows that Dengue fever is the commonest diagnosis made in patients who are detected to have thrombocytopenia. One fifth of patients with platelet count less than 1,00,000/microlitre tend to have bleeding manifestation, commonest being GI bleed, petechial rash and epistaxis. Majority of the bleeding occurs with platelet count less than 10,000. The proportion of patients receiving therapeutic platelet transfusion was higher compared to prophylactic transfusion.

Ademetionine in patients with liver disease: a review

The aim of the present review is to have an in-depth analysis of the published scientific literature relating to the clinical use of ademetionine in various etiologies of liver disease. Literature search was performed using electronic databases like Pubmed/Medline/others to identify studies on ademetionine in patients with intrahepatic cholestasis, alcoholic liver disease, non-alcoholic fatty liver disease, drug induced liver injury and viral hepatitis. Ademetionine has been studied in various etiologies of liver disease with varying dosing and durations. In patients with chronic and alcoholic liver disease, ademetionine was found to be beneficial in improving liver enzyme levels, increasing glutathione levels, improving signs and symptoms of fatigue, pruritus and jaundice. Positive effects of ademetionine therapy have also been documented in multiple studies in patients with non-alcoholic fatty liver disease, with improvements observed in triglyceride, total cholesterol, alanine transaminase and asprtate transaminase levels and ultrasound grading of fatty change. In patients with drug induced liver injury, improvements were observed in liver biochemical markers and symptoms such as pruritus, fatigue and jaundice. Ademetionine has also been studied in patients with viral hepatitis with improvement in laboratory markers and signs and symptoms. Published data suggest that there is clinical evidence to substantiate the use of ademetionine across indications. Its use has resulted in sustained improvement in biochemical markers; signs and symptoms of liver disease has been observed in both acute and chronic liver disease. Further data is warranted through clinical studies to focus on specific end points of therapy areas, in existing and new indications.

Prevalence of genotype D in chronic liver disease patients with occult HBV infection in northern region of India.

Background: Etiology of nearly 30% cases of chronic viral hepatitis remains undetected. Occult HBV infection (OBI) has emerged as an important clinical entity in this scenario. Apart from prevalence and clinical outcome of OBI patients genotype was determined in northern region of India. Materials and Methods: A total of 847 patients with chronic liver disease (CLD) were screened for common viral etiologies and others serological markers of HBV. Amplifi cation of surface, precore and polymerase genes of HBV was performed in patients negative for other etiologies. Genotyping and sequencing of the precore region was performed for OBI cases. Results: Twenty-nine (7.61%) cases of OBI were identifi edof which 9 had chronic liver disease (CHD), 11 liver cirrhosis (LC) and 9 hepatocellular carcinoma (HCC). Majority of OBI cases were detected by amplifi cation of surface gene 26 (89.6%), followed by pre-core gene 12 (41.3%). Their liver functions tests were signifi cantly deranged in comparison to overt HBV cases. IgG anti HBc was present in 8 (27.6%) OBI cases. Mutation was observed in 8 (32%) in pre-core region at nt. 1896 of overt HBV cases. Genotype D was the predominant genotype. In conclusion: OBI in our study was characterized by predominance of genotype D and more severe clinical and biochemical profi le in comparison to overt HBV. IgG anti HBc positivity could be utilized as a marker of OBI. We recommend use of sensitive nested PCR for diagnosis of OBI, amplifying at least surface and precore gene.

Polymorphisms in RAS Guanyl-releasing Protein 3 are Associated with Chronic Liver Disease and Hepatocellular Carcinoma in a Korean Population

RAS guanyl-releasing protein 3 (RasGRP3), a member of the Ras subfamily of GTPases, functions as a guanosine triphosphate (GTP)/guanosine diphosphate (GDP)-regulated switch that cycles between inactive GDP- and active GTP-bound states during signal transduction. Various growth factors enhance hepatocellular carcinoma (HCC) proliferation via activation of the Ras/Raf-1/ extracellular signal-regulated kinase (ERK) pathway, which depends on RasGRP3 activation. We investigated the relationship between polymorphisms in RasGRP3 and progression of hepatitis B virus (HBV)-infected HCC in a Korean population. Nineteen RasGRP3 SNPs were genotyped in 206 patients with chronic liver disease (CLD) and 86 patients with HCC. Our results revealed that the T allele of the rs7597095 SNP and the C allele of the rs7592762 SNP increased susceptibility to HCC (OR=1.55, p=0.04 and OR=1.81~2.61, p=0.01~0.03, respectively). Moreover, patients who possessed the haplotype (ht) 1 ( A-T-C-G) or diplotype (dt) 1 ( ht1/ht1) variations had increased susceptibility to HCC (OR=1.79 ~2.78, p=0.01~0.03). In addition, we identified an association between haplotype1 (ht1) and the age of HCC onset; the age of HCC onset are earlier in ht1 +/+ than ht1 +/- or ht1 -/- (HR=0.42~0.66, p=0.006~0.015). Thus, our data suggest that RasGRP3 SNPs are significantly associated with an increased risk of developing HCC.

Effects of Dexamethasone Treatment on Blood Pressure in Preterm Infants with Chronic Lung Disease (CLD)

PURPOSE: This study was designed to see the BP changes according to the time course and the duration of dexamethasone (DXM) therapy in premature infants with chronic lung disease (CLD). METHODS: We studied 27 chronic lung disease patients treated with DXM in NICU, Chonnam University Hospital from January 1994 to May 1998. Systolic, diastolic, and mean arterial pressure were recorded at three times (8 AM, 4 PM, midnight) daily. Data were analyzed by time peroid : Pre DXM means 14 days before DXM therapy, DXM during the therapy and Post DXM 14 days after the completion of therapy. Of 27 patients, 16 received short-course (7 days), and 11 long-course therapy (42 days). RESULTS: Mean gestational age of the patients was 29.3 (+/-1.5) weeks and the mean birth weight was 1,169 (+/-262) gm. Systolic, diastolic and mean BP were significantly increased during the DXM therapy compared to pre DXM (76+/-7 mmHg vs 67+/-9 mmHg, P<0.001, 44+/-6 mmHg vs 38+/-6 mmHg, P<0.001, 55+/-6 mmHg vs 49+/-7 mmHg, P<0.001, respectively). Even 14 days after the completion of therapy, systolic, diastolic and mean BP were not decreased to the level of pre DXM therapy. The maximal increase of BP was noted on the second day of treatment. When the BP changes were compared according to the duration of therapy, post DXM BP was decreased to the level of pre DXM in short course, but not in long course group with the higher post DXM systolic BP than that of short course group (78+/-11 mmHg vs 69+/-7 mmHg, P<0.05). CONCLUSION: BP significantly increased during DXM therapy, particularily on the second day of treatment. Also our result suggests that we have to watch the BP carefully more than two weeks after the completion of therapy.