Impact of pre-treatment extracellular volume fraction measured by computed tomography on response of primary lesion to preoperative chemotherapy in abdominal neuroblastoma

Abstract Objectives: To retrospectively investigate the impact of pre-treatment Extracellular Volume Fraction (ECV) measured by Computed Tomography (CT) on the response of primary lesions to preoperative chemotherapy in abdominal neuroblastoma. Methods: A total of seventy-five patients with abdominal neuroblastoma were retrospectively included in the study. The regions of interest for the primary lesion and aorta were determined on unenhanced and equilibrium phase CT images before treatment, and their average CT values were measured. Based on patient hematocrit and average CT values, the ECV was calculated. The correlation between ECV and the reduction in primary lesion volume was examined. A receiver operating characteristic curve was generated to assess the predictive performance of ECV for a very good partial response of the primary lesion. Results: There was a negative correlation between primary lesion volume reduction and ECV (r = -0.351, p = 0.002), and primary lesions with very good partial response had lower ECV (p < 0.001). The area under the curve for ECV in predicting the very good partial response of primary lesion was 0.742 (p < 0.001), with a 95 % Confidence Interval of 0.628 to 0.836. The optimal cut-off value was 0.28, and the sensitivity and specificity were 62.07 % and 84.78 %, respectively. Conclusions: The measurement of pre-treatment ECV on CT images demonstrates a significant correlation with the response of the primary lesion to preoperative chemotherapy in abdominal neuroblastoma.

Association of maternal isolated thyroid peroxidase antibody positive in the first trimester with fetal growth / 北京大学学报(医学版)

OBJECTIVE@#To investigate the association of isolated thyroid peroxidase antibody (TPOAb) positive in the first trimester with fetal growth.@*METHODS@#A total of 16 446 pregnant women were included in the birth cohort study, whose last menstrual period was between May 2016 and April 2019 and with singleton pregnancy. Maternal serum samples were collected when they firstly came for prenatal care in the first trimester. The pregnant women were consecutively seen and followed in the hospital and the information of pregnant women was extracted from the electronic medical information system. The pregnant women were divided into isolated TPOAb positive group (n=1 654) and euthyroid group (n=14 792). Three fetal ultrasound examinations were scheduled during the routine prenatal visits at the hospital and were performed by trained sonographers. All fetal growth indicators were quantified as gestational age- and gender- adjusted standard deviation score (Z-score) using the generalized additive models for location, scale and shape (GAMLSS). Fetal growth indicators included estimated fetal weight (EFW), abdominal circumference (AC), biparietal diameter (BPD), femur length (FL) and head circumference (HC). Fetal growth restriction (FGR) was defined as AC or EFW Z-score＜3rd centile based on clinical consensus. Generalized estimating equation (GEE) analysis was applied to assess the association of maternal isolated TPOAb positive with fetal growth. The generalized linear model was further used to analyze the association between isolated TPOAb positive and fetal growth indicator at different gestational ages when the fetal growth indicator was significantly associated with isolated TPOAb positive in the GEE mo-del.@*RESULTS@#The median gestational age at three ultrasound measurements was 23.6 (23.3, 24.1), 30.3 (29.7, 30.9), 37.3 (37.0, 37.7) weeks, respectively. The BPD Z-score was higher in isolated TPOAb positive women, compared with the euthyroid pregnant women after adjustment (β=0.057, 95%CI: 0.014-0.100, P=0.009). The generalized linear model showed the BPD Z-score was higher in the isolated TPOAb positive women at the end of 21-25 weeks (β=0.052, 95%CI: 0.001-0.103, P=0.044), 29-32 weeks (β=0.055, 95%CI: 0.004-0.107, P=0.035) and 36-40 weeks (β=0.068, 95%CI: 0.011-0.125, P=0.020), compared with the euthyroid pregnant women. There was no difference in other fetal growth indicators (EFW, AC, FL and HC) and FGR between the isolated TPOAb positive and euthyroid pregnant women.@*CONCLUSION@#The BPD Z-score was slightly increased in the isolated TPOAb positive pregnant women in the first trimester, while other fetal growth indicators were not changed. The reproducibility and practical significance of this result need to be confirmed.

Expert Consensus on Acute Respiratory Failure in Critically Ill Cancer Patients (2023) / 中国医学科学杂志(英文版)

Objective This consensus aims to provide evidence-based recommendations on common questions in the diagnosis and treatment of acute respiratory failure (ARF) for critically ill cancer patients.Methods We developed six clinical questions using the PICO (Population, Intervention, Comparison, and Outcome) principle in diagnosis and treatment for critical ill cancer patients with ARF. Based on literature searching and meta-analyses, recommendations were devised. The GRADE (Grading of Recommendation Assessment, Development and Evaluation) method was applied to each question to reach consensus in the expert panel. Results The panel makes strong recommendations in favor of (1) metagenomic next-generation sequencing (mNGS) tests may aid clinicians in rapid diagnosis in critically ill cancer patients suspected of pulmonary infections; (2) extracorporeal membrane oxygenation (ECMO) therapy should not be used as a routine rescue therapy for acute respiratory distress syndrome in critically ill cancer patients but may benefit highly selected patients after multi-disciplinary consultations; (3) cancer patients who have received immune checkpoint inhibitor therapy have an increased incidence of pneumonitis compared with standard chemotherapy; (4) critically ill cancer patients who are on invasive mechanical ventilation and estimated to be extubated after 14 days may benefit from early tracheotomy; and (5) high-flow nasal oxygen and noninvasive ventilation therapy can be used as a first-line oxygen strategy for critically ill cancer patients with ARFs. A weak recommendation is: (6) for critically ill cancer patients with ARF caused by tumor compression, urgent chemotherapy may be considered as a rescue therapy only in patients determined to be potentially sensitive to the anticancer therapy after multidisciplinary consultations. Conclusions The recommendations based on the available evidence can guide diagnosis and treatment in critically ill cancer patients with acute respiratory failure and improve outcomes.

Braided stent-assisted coil embolization versus laser engraved stent-assisted coil embolization in patients with unruptured complex intracranial aneurysms

Abstract Purposes: Braided and laser-cut stents both are efficacious and safe for coiling intracranial aneurysms. The study aimed to compare outcomes following braided stent-assisted coil embolization versus laser engraved stent-assisted coil embolization in 266 patients who were diagnosed with unruptured intracranial aneurysms of different types and locations. Methods: Patients with unruptured complex intracranial aneurysms underwent braided (BSE cohort, n = 125) or laser engraved (LSE cohort, n = 141) stent-assisted embolization. Results: The deployment success rate was higher for patients of the LSE cohort than those of the BSE cohort (140 [99%] vs. 117 [94%], p = 0.0142). Seventy-one (fifty-seven percentages) and 73 (52%) were coil embolization procedure success rates of the BSE and the LSE cohorts. Periprocedural intracranial hemorrhage was higher in patients of the BSE cohort than those of the LSE cohort (8 [6%] vs. 1 [1%], p = 0.0142). Four (three percentages) patients from the LSE cohort and 3 (2%) patients from the BSE cohort had in-stent thrombosis during embolization. Permanent morbidities were higher in patients of the LSE cohort than those of the BSE cohort (8 [6%] vs. 1 [1%], p = 0.0389). Higher successful procedures (76% vs. 68%) and fewer postprocedural intracranial hemorrhage (0% vs. 5%) and mortality (0% vs. 5%) were reported for patients of the BSE cohort in posterior circulation aneurysmal location than those of the LSE cohort. Laser engraved stent has fewer problems with deployment and may have better periprocedural and follow-up outcomes after embolization. Conclusions: Braided stent-assisted embolization should be preferred when the aneurysm is present in the posterior circulation.

Polysaccharide of atractylodes macrocephala inhibits the growth of mice in-situ colon cancer HT-29 cell xenograft via activating immune cells / 中国肿瘤生物治疗杂志

@# Objective: To investigate the effect and mechanism of polysaccharide of atractylodes macrocephala (PAM) on the growth of colon cancer cells in mice bearing in-situ colon cancer transplantation tumor. Methods: 1×107 colon cancer HT-29 cells labeled with luciferase were injected into colon serosa of the mice to establish the in-situ colon cancer transplantation tumor model. When the tumor volume reached 230 mm3, the mice were given 30 mg/kg PAM (PAM group) or equal volume of normal saline (Control group) by gavage for 10 consecutive days. The effect of PAM on the growth of colon cancer cells in mice was tested by in vivo tumor imaging technology. The expressions of MHCII and IL-12 in granulocytes, dendritic cells and macrophages, the activation of lymphocytes, and IFN-γ expression in CD4+ and CD8+ cells of tumor tissues were detected by Flow cytometry. Results: PAM significantly inhibited the growth of colon cancer cells in mice bearing in-situ colon cancer transplantation tumor (P<0.01). PAM activated immune cells though increasing the expression levels of MHCII and IL-12 in dendritic cells and macrophages (both P<0.01). PAM significantly increased the frequency of CD8+ cells, NK cells, CD44+/NK cells and CD44+/CD4+ cells in tumor tissues and the number of CD8+ cells and NK cells per unit volume (all P<0.01). PAM significantly increased the IFN-γ secretion of CD4+ and CD8+ cells (both P<0.01), too. Conclusion: PAM inhibits the growth of colon cancer by activating immune cells in tumor tissues of mice bearing in-situ colon cancer transplantation tumor.

AMPK regulates mitochondrial oxidative stress in C2C12 myotubes induced by electrical stimulations of different intensities / 南方医科大学学报

<p><b>OBJECTIVE</b>To study effect of electrical stimulations of different intensities on mitochondrial oxidative stress in C2C12 myotubes and explore the molecular mechanisms.</p><p><b>METHODS</b>After 7 days of differentiation, C2C12 myotubes were subjected to electrical stimulations (15 V, 3Hz, 30 ms) for 60, 120, or 180 min, and the morphological changes of muscular tubes were observed under inverted microscope. The levels of MDA and SOD activity of the cells were detected, and flow cytometry was used to detect mitochondrial reactive oxygen species (ROS) and membrane potential. Western blotting was used to detect the expression of PGC1, AMPK-Ser485, AMPK-Thr172, and AMPK in the cells.</p><p><b>RESULTS</b>No significant changes occurred in the morphology of C2C12 myotubes in response to electrical stimulations. Electrical stimulation for 60 min resulted in significantly increased levels of MDA, AMPK-Ser485 and AMPK-Thr172 in the cells (P<0.05); simulations of the cells for 120 and 180 min caused significantly increased MDA, ROS, mitochondrial ROS, AMPK-Ser485 and PGC1 along with marked reduction of mitochondrial membrane potential (P<0.05).</p><p><b>CONCLUSION</b>Electrical stimulation significantly activates oxidative stress, and a longer stimulation time causes stronger mitochondrial oxidation. AMPK-Thr172 regulates oxidative stress induced by stimulations for a moderate time length, while AMPK-Ser485 and PGC1 function to modulate oxidative stress following prolonged stimulations.</p>

Rictor/mTORC2 regulates blood-testis barrier and spermatogenesis in mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the role of Rictor/mTORC2 in the formation of blood testis barrier (BTB), testicular development, and spermatogenesis.</p><p><b>METHODS</b>Amh Cre positive mice homozygous for rictor loxP with Sertoli cell specific deletion of rictor were obtained by cross breeding Amh Cre mice with rictor loxP mice. The histology of the reproductive organs, seminiferous tubules and epididymis of the transgenic mice was observed with HE staining. The cell subgroups of the germ cells in the seminiferous tubule were detected by flow cytometry with propidium iodide labeling. The expression levels of Ki 67 and separase were detected with immunofluorescence assay, and the expression levels of BTB associated proteins were detected with immunofluorescence and Western blotting.</p><p><b>RESULTS</b>Compared with the control (Amh Cre, rictoror rictor) mice, the mice with Sertoli cell specific rictor deletion showed significantly decreased testicular weight and epididymis weight (P<0.05), significantly increased diploid cells (P<0.01), and decreased haploid cells (P<0.01) but comparable tetraploid cells and similar expression levels of Ki 67 and separase. The mice with rictor knockout also showed aberrant localization of BTB associated proteins, which were scattered over the whole seminiferous epithelium, but the expression levels of the protein remained stable.</p><p><b>CONCLUSION</b>Rictor in testicular Sertoli cells is essential for maintaining BTB integrity and function and ensuring normal spermatogenesis in mice.</p>

Curative effect of compound Xuesaitong combined with Conbercept in the treatment of choroidal neovascularization in pathological myopia / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To analyze the effect of compound xuesaitong combined with conbercept on central macular retinal thickness (CMT), fluorescein fundus angiography and best corrected visual acuity(BCVA) in pathological patients with choroidal neovascularization (CNV).METHODS:A total of 60 cases (60 eyes) with pathological myopia and CNV who were admitted to our hospital during January to October 2016 were enrolled in the study.They were divided into observation group and control group, 30 cases in each group.Observation group was treated with compound xuesaitong combined with conbercept.Control group was treated with conbercept only.Changes of CMT, intraocular pressure (IOP), area of CNV, BCVA and serum vascular endothelial growth factor (VEGF) before surgery, 4 and 12wk after surgery were compared.Complications during follow-up were statistically analyzed.RESULTS: At 12wk after surgery, the total effective rate was 97% in observation group, 80% in control group and the difference was significant (P<0.01).The CMT, IOP, area of CNV and serum VEGF levels were significantly lower or smaller than those before surgery while BCVA was significantly higher than that before surgery (P<0.01).There were no local complications of systematic severe complications.CONCLUSION: Compound xuesaitong combined with conbercept can effectively reduce CMT and IOP, reduce the area of CNV and significantly improve visual acuity.Its anti-VEGF effect is obvious, safe and feasible.

Development of rapid setting-drying device for herbarium specimens and its application in survey of Chinese materia medica resources in Anhui / 中国中药杂志

Herbarium specimens are the basis for the plant classification and indispensable media in teaching, scientific research and resources investigation. They have also played an important role in identifying and producing traditional Chinese medicine. High-quality herbarium specimens shall meet high requirements for integrity, smoothness, color and fabricating efficiency. Therefore, we designed a rapid setting and drying device for herbarium specimens, which could make the herbarium specimens smooth, colorful and not easy to mildew. In this paper, we pointed out the deficiency of traditional methods in making herbarium specimens, and introduced the structure and working principle of the device. Besides, we also discussed the effect of the device in setting and drying herbarium specimens and its application in the fourth national survey of the Chinese material medica resources (CMMR) in Anhui province. As a result, the device provides new ideas for producing herbarium specimens, with a reasonable design, good uniformity, high efficiency, safety and portability, and so is worthy of promotion and application in the national survey of CMMR.

New records of distribution of medical plants in Anhui province / 中国中药杂志

During the fourth national survey of Chinese material medica resources inventory, 9 species of medical plants in Huangshan area of Anhui Province were newly recorded, including Microlepia calvescens, Dryopteris hangchowensis, Fatoua pilosa, Girardinia chingiana, Lecanthus peduncularis, Galium kamtschaticum, Carpesium minus,Cirsium racemiforme, Globba racemosa, which belong to seven families and nine genera. Among these, 3 genera (Girardinia, Lecanthus, Globba) are new geographical distribution in Anhui Province. All of voucher specimens are preserved in ACM. These discoveries enrich the content of flora in Anhui and provide fundamental materials for studying the plants of Anhui.

Historical evolution and development countermeasures of uncommon-territorial herbs / 中国中药杂志

As an important part of Chinese medicinal materials, uncommon-territorial herbs are also the most complex parts in the herbal medicine markets. Through years of investigation on the key markets of Chinese herbal medicine, the meaning of uncommon-territorial herbs, their historical evolution, origin and characteristics were clarified in this paper, and some countermeasures were put forward for its development.

Establishment of a method to detect duck hepatitis B virus covalently closed circular DNA based on rolling circle amplification / 病毒学报

Rolling circle amplification (RCA) is a newly developed experimental technique that can specific ally amplify circular DNA. Since 2008, RCA has been extensively used in hepatitis B virus (HBV) research, such as the amplification of the full-length sequence of the HBV genome, and the analysis of the drug-resistant mutations of HBV covalently closed circular DNA (cccDNA), amongst others. To create an easy assay for the analysis of duck hepatitis B virus (DHBV) cccDNA, this study established an RCA-based method. DHBV cccDNA was amplified from the DHBV DNA samples of duck liver with four pairs of sulfur-modified primers, which were designed according to the highly conserved sequence of DHBV using sera DHBV DNA as the negative control. DHBV cccDNA was detected in the obtained RCA products by the sequencing of RCA amplicons that were amplified with primer pairs on both sides of the gap of DH BV relaxed circular DNA, rather than by digesting RCA products with a restriction enzyme. The liver and sera DHBV DNA samples of 39 ducks infected with DHBV were examined with the RCA-based DHBV cccDNA detection method, and the results showed that while DHBV cccDNA was detected from all 39 liver DHBV DNA samples, no DHBV cccDNA was found in any of the sera DHBV DNA samples. These results suggest that the method established in the study is highly specific and sensitive for the detection of DHBV cccDNA. The establishment of this RCA-based DHBV method for cccDNA detection lays the groundwork for using a DHBV model to study the role of cccDNA in the pathogenesis of hepatitis B and to evaluate the effect of anti-virus therapies.

Cloning and sequence analysis of the DHBV genome of the brown ducks in Guilin region and establishment of the quantitative method for detecting DHBV / 病毒学报

Brown ducks carrying DHBV were widely used as hepatitis B animal model in the research of the activity and toxicity of anti-HBV dugs. Studies showed that the ratio of DHBV carriers in the brown ducks in Guilin region was relatively high. Nevertheless, the characters of the DHBV genome of Guilin brown duck remain unknown. Here we report the cloning of the genome of Guilin brown duck DHBV and the sequence analysis of the genome. The full length of the DHBV genome of Guilin brown duck was 3 027bp. Analysis using ORF finder found that there was an ORF for an unknown peptide other than S-ORF, PORF and C-ORF in the genome of the DHBV. Vector NTI 8. 0 analysis revealed that the unknown peptide contained a motif which binded to HLA * 0201. Aligning with the DHBV sequences from different countries and regions indicated that there were no obvious differences of regional distribution among the sequences. A fluorescence quantitative PCR for detecting DHBV was establishment based on the recombinant plasmid pGEM-DHBV-S constructed. This study laid the groundwork for using Guilin brown duck as a hepatitis B animal model.

Survey of dietary intake of Chinese adults in 2010 / 中华预防医学杂志

<p><b>OBJECTIVE</b>To analyze the status of dietary intake of Chinese adults in 2010.</p><p><b>METHODS</b>In 2010, China Chronic Disease Surveillance was carried out in 31 provinces and Xinjiang Production and Construction Corps. The survey was undertaken in 162 surveillance sites in the national disease surveillance points system by stratified multi-stage cluster random sampling. The participants were local residents (living in the area for more than 6 months) aged 18 years and above in the surveillance sites. A semi-quantitative food frequency questionnaire was used to measure the dietary intake for 97 187 urban and rural adults aged 18 years and above. After complex weighting for the sample, the average daily consumption for major food was analyzed for different genders and regions.</p><p><b>RESULTS</b>After complex weighting, the median of cereal, rice and wheat flour intake for Chinese adults was 375.0 g. The median of livestock meat intake was 57.1 g with western areas (85.7 g) much higher than eastern (57.1 g) and central areas (46.2 g) (χ² = 2054.82, P < 0.01). The median of poultry intake was 7.1 g, eastern areas (10.0 g) higher than western and central areas (both 6.7 g) (χ² = 1210.86, P < 0.01). The median of seafood intake was 14.3 g, eastern (15.0 g) much higher than central (14.3 g) and western areas (7.1 g) (χ² = 5603.37, P < 0.01). The median of daily intake for eggs was 21.4 g, western (14.3 g) lower than eastern and central areas (both 21.4 g) (χ² = 1699.11, P < 0.01). The median of dairy product intake was 42.9 g, eastern (57.1 g) higher than western and central areas (both 35.7 g) (χ² = 196.35, P < 0.01). Overall, 52.8% of urban and rural residents did not meet recommended amount of fruit and vegetable intake. The rate of inadequate intake of fruit and vegetable was higher in rural (55.7%) than urban areas (46.1%, χ² = 929.43, P < 0.01).</p><p><b>CONCLUSION</b>The adult intake of cereal category foods tended to decline in 2010, and the intakes of vegetable and fruits, fishery products as well as dairy products were evidently deficient and regionally varied.</p>

Effect of low-dose fenvalerate on semen quality capacitation in adult mice / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Fenvalerate (FEN) has been demonstrated to be a reproductive toxicant in humans and rodents. However, little is known about whether short-term exposure to low-dose FEN produces reproductive toxicity.</p><p><b>METHODS</b>We administered FEN (0.009 375, 0.1875, 3.750, or 45.00 mg×kg(-1)×d(-1) by gavage for 30 days) to male ICR mice and compared reproductive toxicity parameters between groups receiving different concentrations of FEN. Reproductive toxicity was evaluated by computer-assisted semen quality analysis (CASA), chlortetracycline (CTC) assay, and histopathology.</p><p><b>RESULTS</b>The sperm morphology and testis histology of FEN-exposed mice (all doses) were similar to that in controlling mice. Exposure to FEN at a concentration of 0.1875 mg×kg(-1)×d(-1) decreased sperm path straightness (STR) and linearity (LIN) (both P < 0.05), but had no significant impact on average path velocity (VAP), straight line velocity (VSL), curvilinear velocity (VCL), lateral amplitude (ALH), beat cross frequency (BCF), or progressive motility (MOT). FEN reduced the rate of mouse sperm capacitation in a dose-dependent manner.</p><p><b>CONCLUSION</b>The present results demonstrate that exposure to low-dose FEN for 30 days reduces semen quality and sperm capacitation in adult mice.</p>

Relationship between Tpeak-Tend interval and coronary artery stenosis and effects of percutaneous transluminal coronary angioplasty on Tpeak-Tend / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the relationship between Tpeak-Tend interval (Tpe) and the extent and severity of coronary artery stenosis, and evaluate the effect of percutaneous transluminal coronary angioplasty and stent implantation (PCI) on Tpe in the patients with coronary heart disease (CHD).</p><p><b>METHODS</b>The ECG data were collected from 187 CHD patients undergoing coronary angiography and PCI to evaluate the extend and severity of coronary artery stenosis before and after the interventions.</p><p><b>RESULTS</b>The Tpe of patients with severe stenosis increased significantly as compared with that in patients with moderate stenosis (138.9-/+16.2 ms vs 116.5-/+13.7 ms, P<0.05), and a significant difference was also noted between the moderate stenosis and mild stenosis (86.4-/+12.9 ms) groups (P<0.05). The Tpe decreased significantly in the patients in the order of multi-vessel involvement (140.7-/+17.8 ms), double vessel involvement (118.6-/+14.9 ms), singly vessel involvement (100.5-/+13.2 ms), and stenosis-free (84.3-/+12.4 ms) groups (P<0.05). Tpe was correlated to the extent and severity of coronary artery stenosis (r>0.4). In patients with severe stenosis, the Tpe was significantly reduced at 1 h, 24 h, and 1 week after PCI (115.8-/+14.5, 92.7-/+12.9, and 88.2-/+11.3 ms, respectively, P<0.05).</p><p><b>CONCLUSION</b>The Tpe can reflect the severity and range of coronary artery stenosis, which can be reduced by PCI. Tpe can be a new index for evaluating myocardial ischemia in CHD patients.</p>

Clinical assessment of Tpeak-end interval for prediction of myocardial infarction / 南方医科大学学报

<p><b>OBJECTIVE</b>To assess the value of Tpeak-end interval (Tpe) in predicting myocardial infarction (MI).</p><p><b>METHODS</b>Tpe and Tpeak-end internal after correcting the heart rate (TpeRR) were measured and analyzed in 234 MI patients, who were followed-up for an average of 32 ± 10 months.</p><p><b>RESULTS</b>Clinical events occurred in 45 (19.2%) patients at the end TpeRR of the follow-up. Tpe and of the patients with clinical events were significantly higher than those in patients without the clinical events (P < 0.001). The incidence of clinical events in patients with Tpe > 140 ms were significantly higher than that in patients with Tpe ≤ 140 ms by Kaplan-Meier analysis (P < 0.001). With clinical event as the end point, the proportional hazards rate was 2.48 in univariate COX analysis (P < 0.01). After controlling for risk factors, the hazards rate was 2.66 by multvariate COX regression (P < 0.01).</p><p><b>CONCLUSION</b>Tpe is positively correlated to the prognosis of MI and serves as an new index for predicting the clinical events in MI patients.</p>

Comparative analysis of drug resistant mutations in the reverse transcriptase domain of hepatitis B virus covalently closed circular DNA and the viral relax circle DNA / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To establish a method for detection of reverse transcriptase region of hepatitis B virus (HBV) covalently closed circular DNA (cccDNA), and to compare the pattern and frequency of drug-resistant mutations in the region between intrahepatic HBV cccDNA and serum HBV relax circle DNA (rcDNA).</p><p><b>METHODS</b>HBV DNA were extracted from liver biopsy tissues of 20 patients with chronic hepatitis B. The RT region of HBV cccDNA was amplified by rolling circle amplification (RCA) followed by polymerase chain reaction (PCR) mediated by a pair of primers spanning across the gap region of HBV genome. The RT region of serum HBV rcDNA from the same patient was amplified by nested-PCR. The PCR products were directly sequenced and analyzed by Vector NTI Suite 8.0 and chromaslite 201 software. x2 test was used for statistical significance analysis of drug-resistant mutation occurrences between the HBV cccDNA and rcDNA.</p><p><b>RESULTS</b>The RT regions of HBV cccDNA were successfully amplified from liver tissues of all enrolled patients using the RCA plus PCR assay. Simultaneously, HBV the RT regions of rcDNA were amplified from these patients serum samples. Sequence analysis showed that the drug-resistant mutations were significantly more frequently detected in HBV rcDNA (40%) than in HBV cccDNA (10%) (P<0.05). Different mutational patterns were observed between the HBV cccDNA and rcDNA in a few cases.</p><p><b>CONCLUSION</b>The RCA in combination with PCR is a practical method for the detection of drug-resistant mutation in the RT region of HBV cccDNA. Drug-resistant mutational patterns could be discrepant between HBV cccDNA and rcDNA.</p>

Analysis and recombinant construction of HBV the reverse transcriptase gene with drug-resistant mutations from 40 patients with chronic hepatitis B / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To analyze genetic mutation associated with drug resistance in the reverse transcriptase (RT) domain of HBV from 40 patients with chronic hepatitis B, and to construct mutant RT gene recombinant vectors for drug-resistant phenotypic analysis.</p><p><b>METHODS</b>HBV DNA was extracted from sera of the 40 patients receiving anti-HBV nucleot (5) ide analogue. The complete RT domain-encoding gene was amplified by nested PCR, and then cloned into pGEM-T-easy vector. Three to Five clones were randomly selected for DNA sequencing. Data were analyzed by UNASTAR software. The pTriEx-HBV (C) 1.1 expression vectors were constructed by replacing the 1250-hp Xho I/Nco I fragments containing complete RT domain from individual patients samples.</p><p><b>RESULTS</b>All samples were detected with drug-resistant mutations associated with lamivudine, adefovir, and entacavir singly or in combination. Ninety-six mutant RT genes were cloned into pGEM-T-easy vector, from which 40 major mutant RT genes were replaced into pTriEx-HBV (C) 1.1 expression vectors. The construction was confinned to be successful by verifying mutation existence using DNA sequencing, and detectable HBsAg and HBeAg in the cell supernatant after transfecting recombinant expression vectors into Huh7 cells.</p><p><b>CONCLUSION</b>The analysis of drug-resistant mutation and the construction of mutant-recombinant expression vectors were successfully implemented using the samples frum clinical patients. The work lays a foundation for drug-resistant phenotypic analysis of HBV mutants.</p>

Chronological changes of pathology in cerebellar degeneration in rats with methylmercury induced toxication / 第二军医大学学报

Objective:To explore the pathology and pathogenesis of cerebellar injuries induced by methylmercury chloride(MMC) toxication in rats. Methods:Rats were given MMC(4 mg·kg-1·d-1) consecutively and sacrificed on days 11, 15, 18 and 21. Pathological changes of the cerebellum were observed by histo-immunopathology; in situ staining was performed for DNA strand breaks in cerebellar granule cells by TUNEL technique; and the ultrastructures were observed by electron microscope. Results:On day 18, sparse TUNEL positive granular cells were observed mainly in deep lamina adjacent to the white matter. On day 21, apoptotic cells markedly increased and granule cells decreased with well-preserved Purkinje cells. Immunostaining with MRF-1 and GFAP demonstrated severe microgliosis and astrocytosis. On day 18, electron microscopy demonstrated that the nuclei of MMC-treated animals were shrunken and displayed increased electron density, and some homogeneously dense nuclear chromatin with tear-drop features, which were compatible with the apoptotic changes. Conclusion:These results indicate that the pathological changes in the cerebellum in this subacute MMC intoxication model resemble human cases, and the degeneration of granule cells is apoptosis.