Predictive values of ALBI and EZ-ALBI scores for early survival of recipients with liver failure after liver transplantation / 器官移植

Objective To evaluate the predictive values of albumin-bilirubin (ALBI) and easy albumin-bilirubin (EZ-ALBI) scores for early survival (postoperative 3 months) of recipients with liver failure after liver transplantation. Methods Clinical data of 137 recipients diagnosed with liver failure and underwent liver transplantation were retrospectively analyzed. The optimal cut-off values of preoperative ALBI, EZ-ALBI and MELD scores to predict early survival of recipients with liver failure after liver transplantation were determined by the area under the receiver operating characteristic (ROC) curve. The risk factors of early death of recipients with liver failure after liver transplantation were identified by univariate and multivariate Cox regression analyses. The effects of different ALBI and EZ-ALBI levels upon early prognosis of recipients with liver failure after liver transplantation were analyzed. Results The optimal cut-off values of ALBI, EZ-ALBI and MELD scores were 0.21, -19.83 and 24.36, and the AUC was 0.706, 0.697 and 0.686, respectively. Univariate Cox regression analysis showed that preoperative alanine aminotransferase(ALT)≥50 U/L, aspartate aminotransferase(AST)≥60 U/L, ALBI score≥0.21 and EZ-ALBI score≥-19.83 were the risk factors for early postoperative death of recipients with liver failure after liver transplantation (all P < 0.05). Multivariate Cox regression analysis demonstrated that preoperative ALBI score≥0.21 was an independent risk factor for early postoperative death of recipients with liver failure after liver transplantation (P < 0.05). According to the optimal cut-off value of ALBI score, the early survival rates in the ALBI < 0.21 (n=46) and ALBI≥0.21(n=91) groups were 93.5% and 64.8%, and the difference was statistically significant (P < 0.05). According to the optimal cut-off value of EZ-ALBI score, the early survival rates in the EZ-ALBI < -19.83(n=60) and EZ-ALBI≥-19.83(n=77) groups were 88.3% and 63.6%, and the difference was statistically significant (P < 0.05). Conclusions Preoperative ALBI score is of high predictive value for early survival of recipients with liver failure after liver transplantation, which could be utilized as a reference parameter for selecting liver transplant recipients.

Índice de necesidad de tratamiento periodontal en embarazadas y no embarazadas que asisten a la maternidad provincial de Córdoba, Argentina / Periodontal index for treatment needs in pregnant and non-pregnant women attending the provincial maternity hospital in Córdoba, Argentina

Objetivo: Comparar la salud periodontal de embaraza- das y no embarazadas mediante la aplicación del Índice de Periodontal Comunitario (IPC). Materiales y métodos: Se realizó un estudio ob- servacional de corte transversal. Se reclutaron 100 mujeres embarazadas (EMB) y 50 no embarazadas (NoEMB) que concurrieron al Hospital Materno Provincial de la Ciudad de Córdoba, Dr. Raúl F. Lucini. En todas se registró el IPC con la sonda periodontal WHO 621 en los 6 sextantes de la boca. Los datos se analizaron con el software Infostat/SP; el nivel de significación establecido fue de P <0,05. Resultados: El 70% de las pacientes presentó edades de entre 18 y 25 años. En las EMB el código 3 del IPC fue el más frecuente presente en 240 sextantes (40,1%) y en las NoEMB el código 2 fue el más frecuente con 39 sextantes (43%). A ambos grupos de estudio les corresponde el trata- miento de instrucción de higiene bucal, instrumentación supra y/o subgingival, y/o regularización de obturaciones. Conclusiones: El código 3 fue el más frecuente entre las EMB, a quienes les corresponde un Código de tratamiento periodontal (CTP) 2; las NoEMB presentaron un IPC de 1 y 2 como los más frecuentes y se vinculan con un CTP 1 y 2. Nos encontramos frente a una situación clínica periodontal posible de resolver con terapia básica que puede ser realizada por odontólogos generalistas (AU)

Aim: To compare the periodontal health of pregnant and non-pregnant women by applying the Community Periodontal Index (CPI). Materials and methods: In an observational, cross-sec- tional study, 100 pregnant women (PREG) and 50 non-preg- nant women (NonPREG) were recruited at the Dr. Raúl F. Lu- cini Provincial Maternity Hospital in Córdoba City. The CPI was determined in the 6 sextants of the mouth using a WHO 621 periodontal probe. The data were analyzed with Infostat SP software. P <0.05 was considered significant. Results: 70% of the patients were 18 to 25 years old. In the PREG group, CPI Code 3 was the most frequent, present in 240 sextants (40.1%), while in the non-PREG group, CPI Code 2 was the most frequent, with 39 sextants (43%). Treat- ment needs in both study groups are oral hygiene instruction, supra- and/or subgingival instrumentation, and/or correction of plaque retentive margins. Conclusions: Code 3 was the most frequent among preg- nant women, which corresponded to Periodontal Treatment Code (CTP) 2. CPI 1 and 2 were the most frequent in non-pregnant women, corresponding to CTP 1 and 2. This periodontal clinical condition can be treated with initial dental hygiene therapy, which can be performed by general dentists (AU)

Clinical progress of diagnostic and evaluation criteria for liver transplantation in patients with severe liver disease / 器官移植

Liver transplantation, although recognized as the only effective radical treatment for severe liver disease, might be accompanied by high surgical risks, high perioperative mortality and high postoperative complications. Considering the shortage of donor liver and related surgical risks, it is necessary to strictly control the indication of operation and the opportunity of transplantation. Therefore, accurate diagnosis and comprehensive evaluation of the condition of patients with severe liver disease to be treated by liver transplantation is an important part in determining the treatment plan. At present, there are many evaluation criteria for severe liver disease. In addition to the classic ChildTurcotte-Pugh (CTP) score and model for end-stage liver disease (MELD) score, many other evaluation criteria have also been developed. All transplant centers have their own choices and thus there is no uniform diagnostic criterion, with disputes among various criteria, which is exactly what this paper aims to summarize.

Role of lipid profile in assessment of severity of cirrhosis

Background: Cirrhosis is defined anatomically as a diffuse process with fibrosis and nodule formation. It is the result of the fibrogenesis that occurs with chronic liver injury. For reduced liver biosynthesis capacity, low level of serum cholesterol, low density lipoprotein (LDL) and high density lipoprotein (HDL) is usually observed in the chronic liver disease. Due to the high prevalence of chronic liver disease in our country we have conducted this study to determine role of lipid profile in a patient with cirrhosis and to assess its relationship to the severity of cirrhosis.Methods: In this cross-sectional study, patients classified in 3 groups as per CTP classification for severity of cirrhosis. Serum lipid profile was observed in these patients. The primary aim was to assess changes in various parameters of lipid profile and its relationship with severity of liver cirrhosis.Results: About 74 cirrhotic patients were enrolled, 20 in class A, 25 in class B and 29 in class C. serum lipid profile was observed in these patients. Serum cholesterol and HDL cholesterol were decreased with increasing severity of cirrhosis. Serum triglyceride level increases with progression of cirrhosis and very low density lipoprotein (VLDL) level has no correlation with severity of cirrhosis.Conclusions: Serum cholesterol and HDL level decreases with progression of cirrhosis. In future serum lipid profile can be used in classification criteria for assessing severity of liver cirrhosis.

Infliximab biosimilar CT-P13 is interchangeable with its originator for patients with inflammatory bowel disease in real world practice

BACKGROUND/AIMS: An interim analysis of post-marketing surveillance of CT-P13, an infliximab biosimilar, was performed to evaluate its safety and efficacy in Japanese patients with inflammatory bowel disease.METHODS: Patients were prospectively enrolled between November 2014 and March 2017, after the launch of CT-P13 in Japan, and case report forms of patients followed for at least 4 months were analyzed as of July 2018.RESULTS: Of 523 patients in the analysis set, 372 remained on CT-P13 therapy, while 54 (20.2%) of 267 patients with Crohn’s disease, and 97 (37.9%) of 256 patients with ulcerative colitis were withdrawn during follow-up. A total of 144 adverse drug reactions (ADRs) were reported in 106 patients (20.3%). Infusion reaction was the most frequent ADR observed in 49 patients (9.4%). Efficacy parameters decreased immediately after the start of treatment in naïve patients to anti-tumor necrosis factor-α antibody. In the patients switched from originator infliximab for nonmedical reasons, the decreased parameters due to proceeded treatment with the originator were maintained in low ranges, and the treatment continuation rate was high with low ADR incidence. In contrast, in patients switched for medical reasons such as adverse event or loss of response, the incidence of ADRs was high. However, the efficacy parameters were improved, and the treatment continuation rate was not significantly different from that of the naïve patient group.CONCLUSIONS: In this interim analysis, CT-P13 was comparable to the originator infliximab with respect to ADRs and efficacy, and is therefore considered to be a cost-efficient interchangeable biosimilar for Japanese patients with inflammatory bowel disease.

Efficacy of sequential endoscopic variceal ligation plus endoscopic variceal sclerotheropy and the factors associated with the prognosis / 中国内镜杂志

Objective To explore the efficacy of sequential endoscopic variceal ligation plus endoscopic variceal sclerotheropy and the factors associated with the prognosis.Methods 106 cases with esophageal varices in control group was treated with drugs alone;study group had 113 cases, was given endoscopic therapy add drugs, The study group randomly divided into two groups, one was treated with endoscopic variceal ligation all the time (EVL group), another was treated with sequential endoscopic variceal ligation plus endoscopic variceal sclerotheropy (sequential group). After the treatment, the rate of removal of varicose veins, the rate of rebleeding, the number of total treatment,mortality and intra-operative complications and postoperative complications were compared. And compared rebleeding rate and mortality with the control group and study groups with different CTP and MELD, analyze the factors of prognosis, and evaluate their prognostic value.Results Rebleeding, rate in control group, EVL group and sequential group were 41.51%,10.53% and 10.64%,sequential group was significantly better than control group (P = 0.000); mortality in control group, EVL group and sequential group were 15.09%, 5.26% and 2.13%, sequential group was also significantly better than control group (P = 0.001); rate of recurrence in EVL group and sequential group within half a year were 73.68% and 44.68%, sequential group was significantly better than EVL group (P = 0.021). In all control group and EVL group and the sequential group, rebleeding rate and mortality of the liver function Child-Turcotte-Pugh (CTP) class C was significantly higher than that of calss A; In MELD model, AUC area under the ROC curve of rebleeding rate in control group and sequential group were 0.944 and 0.851, mortality of the two groups were 0.881 and 0.984, while the rate of recurrence in the EVL group and sequential group were respectively 0.914 and 0.765, the MELD score has the important value to the prediction of rebleeding and death.Conclusion The rebleeding rate and mortality in cirrhotic patients with esophageal varices treated with sequential endoscopic variceal ligation plus endoscopic variceal sclerotheropy were significantly decreased and the recurrence rate was lower than that of the patients with endoscopic variceal ligation all the time. Liver function Child-Turcotte-Pugh (CTP) score and the MELD score have important value in prediction of rebleeding and death, ligation and sclerosing sequential therapy can significantly reduced rebleeding and mortality in CTP class B and C, and improve the MELD threshold of rebleeding and death.

Current status of biosimilars in the treatment of inflammatory bowel diseases

Introduction of biological therapies have led to dramatic changes in the management of debilitating immune-mediated inflammatory bowel diseases (IBD) including ulcerative colitis and Crohn's disease. However, the long term use of these agents may be very expensive, placing a significant burden on National Healthcare Systems. The development of first biosimilar to infliximab, CT-P13 (Remsima; Celltrion Inc., Incheon, Korea and Inflextra; Hospiral, Lake Forest, Illinois, USA) has become another way to decrease the medical care cost and increase patient treatment option, but, actual equivalence of efficacy and safety of CT-P13 was investigated in rheumatic diseases only. The extrapolation of outcome from rheumatic trials to IBD and the interchangeability of CT-P13 with infliximab have come to be a matter of concern. Two recent retrospective studies reported the similarity of CT-P13 in terms of efficacy and safety. Infliximab biosimilars may be promising new treatment options for IBD patients, however, well-designed, prospective randomized non-inferiority trials should be needed to confidently integrate infliximab biosimilars into IBD treatment.

Chloroplast localization of Cry1Ac and Cry2A protein- an alternative way of insect control in cotton

BACKGROUND: Insects have developed resistance against Bt-transgenic plants. A multi-barrier defense system to weaken their resistance development is now necessary. One such approach is to use fusion protein genes to increase resistance in plants by introducing more Bt genes in combination. The locating the target protein at the point of insect attack will be more effective. It will not mean that the non-green parts of the plants are free of toxic proteins, but it will inflict more damage on the insects because they are at maximum activity in the green parts of plants. RESULTS: Successful cloning was achieved by the amplification of Cry2A, Cry1Ac, and a transit peptide. The appropriate polymerase chain reaction amplification and digested products confirmed that Cry1Ac and Cry2A were successfully cloned in the correct orientation. The appearance of a blue color in sections of infiltrated leaves after 72 hours confirmed the successful expression of the construct in the plant expression system. The overall transformation efficiency was calculated to be 0.7%. The amplification of Cry1Ac-Cry2A and Tp2 showed the successful integration of target genes into the genome of cotton plants. A maximum of 0.673 µg/g tissue of Cry1Ac and 0.568 µg/g tissue of Cry2A was observed in transgenic plants. We obtained 100% mortality in the target insect after 72 hours of feeding the 2nd instar larvae with transgenic plants. The appearance of a yellow color in transgenic cross sections, while absent in the control, through phase contrast microscopy indicated chloroplast localization of the target protein. CONCLUSION: Locating the target protein at the point of insect attack increases insect mortality when compared with that of other transgenic plants. The results of this study will also be of great value from a biosafety point of view.

Structural characterization of the polysaccharide CTP3-E from Crepis turczaniowii C. A. Mey / 中国药学杂志

OBJECTIVE: To characterize the chemical structure of a homogenous polysaccharide CTP3-E purified from Crepis turczaninowii C. A. Mey. METHODS: Sugar composition analysis, methylation analysis, IR and NMR (Dept, HMBC, HMQC, etc.) were used to elucidate the structure. RESULTS: The molecular weight of CTP3-E was 12182. CTP3-E was composed of mannose, rhamose, glucuronic acid, galacturonic acid, glucose, galactose, and arabinose. CONCLUSION: CTP3-E is a new acidic heteropolysaccharide and was isolated from this medicinal plant for the first time.

Clinical research on the correlation of liver function CTP classification and MELD score of post-hepatitis B cirrhosis between different TCM syndromes / 国际中医中药杂志

Objective To explore and identify the correlation of CTP classification,MELD score of patients with cirrhosis after hepatitis between different TCM syndromes.Methods For 135 patients comply with hepatitis B cirrhosis standard,standardized TCM syndrome differentiation and laboratory tests is executed so as to analyze the correlation of CTP classification,MELD score of patients with cirrhosis after hepatitis between different TCM syndromes.Results The liver function CTP classification in liver Qi stagnation syndrome and dump-heat accumulation syndrome patients mainly scores to Class A,and the MELD score less than 10 points-based.The liver function CTP classification in Yin deficiency of Liver and kidney,Yang deficiency of spleen and kidney and blood stasis syndromes mainly scores to Class B-C,and the MELD scores more than 10 points basically.Yin deficiency of Liver and kidney,Yang deficiency of spleen and kidney and blood stasis syndromes take majority in advanced cirrhosis patients compared with liver Qi stagnation syndrome and dump-heat accumulation syndrome take majority in early stage cirrhosis patients.Conclusion Matches the traditional view in TCM that form Qi stagnation,to damp heat-water stopped,to blood stasis-Yang and Yin deficiency is the regular pathogenesis evolution pattern of post-hepatitis cirrhosis.

Comparison of CTP, MELD, and MELD-Na Scores for Predicting Short Term Mortality in Patients with Liver Cirrhosis / 대한소화기학회지

BACKGROUND/AIMS: MELD-Na (model for end-stage liver disease with incorporation of serum sodium) was suggested to provide better survival prediction than MELD alone for patients with end stage liver disease. However, there is no data verifying the usefulness of MELD-Na for predicting short term mortality of cirrhotic patients in Korea. This study was aimed to determine whether MELD-Na would be more accurate in predicting short term mortality than other scoring systems such as Child-Turcotte-Pugh (CTP) or MELD. METHODS: Data from 355 patients admitted due to liver cirrhosis were retrospectively reviewed. The cumulative survival rates were obtained. Prediction of mortality rate for three months and one year were analyzed using the area under the receiver's operating characteristics curve (AUC). RESULTS: One hundred patients (28%) died during the study period. All of the three systems showed significant differences in the cumulative survival rate according to the scores on admission (p0.05), and the AUC of each score system for death within one year were 0.792, 0.800, and 0.831, respectively (p>0.05). The AUC of MELD-Na in predicting short term death were the highest, although it was not statistically significant. Multivariate analysis showed that only MELD-Na was significantly related to three-month mortality (p=0.012). CONCLUSIONS: MELD-Na is more appropriate in predicting short term mortality, but larger scale studies are needed to confirm the superiority of MELD-Na to MELD and CTP in patients with liver cirrhosis.

Usefulness of delta MELD/month for Prediction of the Mortality in the First Episode of Variceal Bleeding Patients with Liver Cirrhosis: Comparison with CTP, MELD score and delta CTP/month / 대한간학회지

BACKGROUND/AIMS: There are many models for predicting prognosis of liver cirrhosis including Child Turcotte Pugh (CTP), the model for end-stage liver disease (MELD) score and its changes over time (delta CTP and delta MELD/month). We investigated the ability of these models to predict the mortality of liver cirrhosis patients with the first episode of variceal bleeding and which model can be usefully applied in practice. METHODS: Seventy-one liver cirrhosis patients hospitalized for the first episode of esophageal variceal bleeding were retrospectively analyzed. The predictive power of initial CTP, MELD score, delta CTP and delta MELD/month was compared through c-statistics and multiple logistic regression. RESULTS: All of the prognostic predictors measured higher in patients who survived than in those who died. The area under the receiver operating characteristic (ROC) curve for delta MELD/month in 6 months was 1, a higher value than 0.81 for initial CTP, 0.75 for initial MELD, and 0.84 for delta CTP/month; the area of delta MELD/month in 12 months was 0.81, also showing a higher value than others. delta MELD/month >0.27 was a strong significant prognostic predictor in 6 (odds ratio: 40.1, p=0.001) and 12 months (odds ratio: 14.1, p<0.001). Only the delta MELD/month was an independent prognostic predictor with a risk ratio of 1.604 (95% CI: 1.119-2.302, p=0.01) in 6 months and 1.627 (95% CI: 1.294-2.047, p<0.001) in 12 months. CONCLUSIONS: The delta MELD/month is superior to initial CTP, MELD and delta CTP/month to predict 6 and 12 months mortality in liver cirrhosis patients with the first episode of variceal bleeding.

Usefulness of CT Perfusion in the Postoperative Evaluation of Ruptured Aneurysm / 대한뇌혈관외과학회지

OBJECTIVE: Our goal was to evaluate the usefulness of CT perfusion (CTP) in early detection of the post operative cerebral ischemia, alteration of treatment modality and patient prognosis in cerebral aneurysm patients. METHODS: 24 patients who underwent either surgical operation or endovascular coiling for ruptured aneurysms were selected. All patients undertook an angiogram, conventional CT, and CTP scan immediately following surgical operation or endovascular coiling. All patients performed a CT 2 weeks after treatment to evaluate possible development of a cerebral infarction. Postoperative CT results of patients with abnormal postoperative CTP scan findings were compared, and these results were compared with the CT results and clinical symptoms of patients who developed infarction or not. RESULTS: Of the 24 patients evaluated, 11 patients showed abnormal findings on CTP. 9 patients were diagnosed with cerebral infarction through a CT scan done 2 weeks after treatment; all exhibited abnormal CTP results immediately after treatment. Abnormal CTP findings were divided into two groups; patients with abnormal CBF and MTT maps, but with normal CBV maps, and patients with abnormal CBF, CBV and MTT maps. A correlation was seen between abnormality on CBV maps and cerebral infarction. Patients with abnormal CTP findings also exhibited poorer prognostic value. CONCLUSION: Postoperative CTP in ruptured aneurysm patients is a very useful and objective tool in evaluating abnormal cerebral hemodynamics. The CBV map of CTP is the most precisely predictable value of postoperative patient's status and alteration of treatement modality.

The Anaylsis of Mortality Rate According to CTP Score and MELD Score in Patients with Liver Cirrhosis / 대한간학회지

BACKGROUND/AIMS: The Model for End-Stage Liver Disease (MELD) consists of serum bilirubin and creatinine levels, International Normalized Ratio (INR) for prothrombin time, and etiology of liver disease. The MELD score is a reliable measurement of mortality risk and is suitable for a disease severity index in patients with end-stage liver disease. We examined the validity of the MELD as a disease severity index for patients with end-stage liver disease. METHODS: We investigated the 379 patients with liver cirrhosis hospitalized between January 1995 and May 2001. We retrospectively reviewed the hospital records to verify the diagnosis of cirrhosis and to collect exact patient information about their demographic data, portal hypertensive complications and laboratory data. The ability to classify patients with liver cirrhosis according to their risk of death was examined using the concordance c-statistic. RESULTS: The MELD score performed well in predicting death within 3 months with a c-statistic of 0.73 with etiology and 0.71 without etiology. The significant clinical, laboratory variables on 3 month survival in patients with liver cirrhosis are serum bilirubin, ascites and hepatic encephalopathy. The addition of portal hypertensive complications to the MELD score did not improve the accuracy of the MELD score. CONCLUSIONS: The MELD score is a useful disease severity index for the patients with end-stage liver disease and provides reliable measurement of short term survival over a wide range of liver disease severity and diverse etiology.

Effects of Fas protein expression on neural apoptosis in hippocampus after traumatic brain injury in rats / 第三军医大学学报

Objective To study the mechanisms of nerve cell apoptosis after traumatic brain injury and the influences of CTP on rats after brain trauma. Methods The model of severe closed traumatic brain injury (TBI) was used. A total of 300 Wistar rats were divided randomly into TBI group, CTP treatment group, and sham operation group. The Fas protein expression and neural apoptosis in each group were observed at 3, 6, 12, 24, 48, 72, 168, and 336 h after TBI. At the same time, the rest 12 rats were employed as the normal controls. Immunohistochemistry and TUNEL staining were used to observe the neural apoptosis and the expression of Fas protein in hippocampus of rats after TBI. Results Increased expression of Fas protein and neural apoptosis were found in the rat hippocampus after TBI. Conclusion The increased expression of Fas protein may result in neural apoptosis after TBI. CTP can reduce the expression of Fas protein and neural apoptosis.

Determination of Intracellular Ara-CTP Level by RP-HPLC / 中国药房

OBJECTIVE:To establish a simple and rapid ion-pair-RP-HPLC method for the determination of intracellular concentration of ara-CTP.METHODS:Mononuclearcells were separated from the whole blood,centrifugated and injected for determination on Symmetry C18 column with the mobile phase consisted of 0.1 mol?L-1 KDP buffer solution and 0.01 mol?L-1 tetrabutylammonium hydrogen sulfate(pH=2.7)at a flow rate of 1.0 mL?min-1 with the UV wavelength set at 278 nm.RESULTS:The linear range of ara-CTP was 0.35-9.12 ?g?mL-1(r=0.998 6)with its detection limit at 0.35 ?g?mL-1.The recoveries of ara-CTP at low,middle and high concentrations,and the intra-day and inter-day RSDs were all up to the standards.CONCLUSION:The established method is simple,rapid,stable,reproducible and applicable for the monitoring of intracellular concentration of ara-CTP.

Detection of CTP gene expression in the asexual erythrocytic stages of Plasmodium falciparum (FCC1 /HN) by reverse transcriptase- polymerase chain reaction and construction of eukaryotic expression vector of CTP gene / 热带医学杂志

Objectve To detect whether the CTP(phosphocholine cytidylyltransferase) gene was expressed in the asexual erythrocytic stages of Plasmodium falciparum (FCC 1/HN )by using the RT - PCR and to construct eukaryotic expression vector of CTP. Method The erythrocytic stage parasites of Plasmodium falciparum were cultured as described by Trager and Jensen. RNA from erythrocytic stage parasite was extracted by using Trizol reagent. The complete genes coding for CTP gene isolates FCCI/HN were amplified by reverse transcriptase -polymerase chain reaction(RT- PCR). CTP gene was cloned into eukaryotic expression vector pcDNA3. Results CTP encoding gene was amplified from the erythrocytic stages of Plasmodiumfalciparum (FCC 1/HN) and eukaryotic expression vector of CTP was constructed. Conclusion CTP gene was expressed in the erythrocytic stages of Plasmodium falciparum (FCC 1/HN) and eukaryotic expression vector of CTP was successfully constructed.

Effect of 3,3',5-trliodothyronine on the expression of Go?in primary cultured neurons of rats / 中华内分泌代谢杂志

Objective To investigate the effect of triiodothyronine on the expression of alpha subunit of C protein Co (Coa) gene in cultured rat cerebral cortex neurons. Methods Primary cultured neurons of rat were prepared from the cerebral cortex at embryonic day 19. The neurons were cultured in DMEM supplemented with 15% fetal calf serum. After 7 days, the neurons were cultured in different media; DMEM supplemented with 15% fetal calf serum (group A), DMEM supplemented with 15% fetal calf serum stripped of thyroid hormone (group B), 0.5nmol/L T3-containing DMEM supplemented with 15% fetal calf serum stripped of thyroid hormone (group C) , 5nmol/L T3-containing DMEM supplemented with 15% fetal calf serum stripped of thyroid hormone (group D) and 50nmol/L T3-containing DMEM supplemented with 15% fetal calf serum stripped of thyroid hormone (group E). The neurons were cultured for another 7 days and total RNA was extracted. Goa mRNA leves were measured by competitive RT-PCR. Results As compared with group A, Coa mRNA leves in group B, D and E were low (P 0.05). Conclusion T, has dual effects on the expression of Coa gene in cultured rat cerebral cortex neurons. T3 down-regulated Coa mRNA in cultured neuron. However, in the absence of T3, Goa mRNA was also very low. Thyroid hormone may exert their action on brain development by regulating the the expression of Goa gene.

Properties of Hepatitis B Virus Associated DNA Polymerase

The nature of hepatitis B virus (HBV) particle associated DNA polymerase was studied in relation to various enzyme inhibitors including antiviral agents. HBV DNA polymerase required high concentration of MgCl2(> 30 mM) and neutral pH for its full activity. p-chloromercuribenzoate was a strong inhibitor (85% inhibition at 1 mM) but N-ethylmaleimide had much less inhibitory effect (20% inhibition at 10 mM). Phosphonoformic acid showed the greatest inhibitory effect on HBV-DNA polymerase (almost complete inhibition at 100 microM) among phosphocompounds tested. Adenine arabinoside triphosphate (ara-ATP) and cytosine arabinoside triphosphate (ara-CTP) were competitive inhibitors with respect to their respective deoxyribonucleoside triphosphate (dATP and dCTP). Ara-CPT was a stronger inhibitor of HBV-DNA polymerase compared to ara-ATP. Ki values for ara-ATP and ara-CTP were 15.0 microM and 11.7 microM , respectively. HBV-DNA polymerase is characteristic in its ionic requirements and susceptibilities to certain inhibitors.

Construction and eukaryotic expression of recombinant plasmid ?hCG-CTP/pcDNA3 / 第三军医大学学报

Objective To clone and express the gene fragment of human chorionic gonadotrophin-? carboxyl terminal peptide (?hCG-CTP) and to explore the possibility of mouse immunocontraception with ?hCG-CTP gene fragment. Methods ?hCG-CTP gene fragment was chemically synthesized. ?hCG-CTP DNA fragment was digested with EcoRⅠ and BamHⅠ together, and then ligated to eukaryotic expression vector pcDNA3 which was also digested under the same condition. Product of ligation reaction was transformed into Escherichia coli DH5?. Recombinant plasmid pcDNA3/?hCG-CTP was then transfected into COS-7 cells with liposome. The expression of ?hCG-CTP gene fragment in mammalian animal cells was detected by immunohistochemistry. Results Double restriction enzyme digestion and subsequent sequencing of recombinant plasmids confirmed the correctness of the recombinant plasmids. Recombinant plasmid pcDNA3/?hCG-CTP was transfected into COS-7 cells with liposome, and transient expression of ?hCG-CTP protein was obtained. Conclusion Recombinant plasmid pcDNA3/?hCG-CTP which can express ?hCG-CTP gene in mammalian animal cells has been constructed correctly. The expressed product ?hCG-CTP can be recognized by antibody against ?hCG.