Whole-transcriptome sequencing and bioinformatics analysis of calcified outer-layer tissue from cystic echinococcosis.

Echinococcosis is a common zoonotic disease in livestock; the type with the highest incidence is cystic echinococcosis (CE). In clinical management, patients with CE of the liver in which the cyst wall is calcified have been found to have better prognoses than those without calcification. In this study, we collected calcified and uncalcified cyst wall tissue from patients with hepatic CE and observed significant changes in the expression of 2336 messenger ribonucleic acids (mRNAs), 178 long noncoding RNAs (lncRNAs), 210 microRNAs (miRNAs), and 33 circular RNAs (circRNAs) using high-throughput sequencing (HTS). Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses of differentially expressed RNAs (DERNAs: DEmRNAs, DElncRNAs, DEmiRNAs, and DEcircRNAs) were performed to explore these RNAs' potential biological functions and signaling pathways. Ultimately, the results of hematoxylin and eosin (H&E) and terminal deoxynucleotidyl transferase deoxyuridine triphosphate (dUTP) nick end labeling (TUNEL) staining confirmed the correlation between calcification and apoptosis of the cyst wall. In summary, this study was an initial exploration of the molecular-biological mechanism underlying spontaneous calcification of the hydatid cyst wall, and it provides a theoretical basis for exploring new targets for drug treatment in CE.

Correlations between the femoral neck osteotomy angle and radiologic and clinical outcomes analyzed in patients undergoing total hip replacement with metaphyseal fixation.

OBJECTIVE: This study will explore whether the femoral neck osteotomy angle (FNOA) has an effect on hip anatomical functional reconstruction and clinical outcomes after total hip arthroplasty (THA). PATIENTS AND METHODS: The study included 254 patients (296 hips) who underwent primary total hip arthroplasty using the same uncemented short stem (Tri-Lock BPS) between December 2018 and December 2019. Correlations between FNOA and the radiologic and clinical outcomes of patients were analyzed. RESULTS: Patients were divided into 3 groups according to different FNOA. FNOA ≤50° is Group A, 50°< FNOA <55° Group B, and FNOA ≥55° Group C. There were significant differences among the three groups in distal D1 (p=0.029), sitting proud (SP) (p<0.001), varus and valgus alignment (p<0.001), FO (p=0.001), and caput-collum-diaphysis angle (CCD) (p<0.001). There were significant differences in the incidence of complications among the three groups (p<0.007). There was a significant linear correlation with D1 (B=0.005, CI=0.002 to 0.008, p=0.004), SP (B=-0.266, CI=-0.286 to 0.166, p<0.001), the femoral stem varus-valgus alignment angle (B=-0.359, CI=-0.422 to -0.297, p<0.001), femoral offset (FO) (B=-0.500, CI=-0.795 to -0.205, p=0.001), and CCD (B=0.696, CI=0.542 to 0.849, p<0.001). In logistic regression analysis, inappropriate FNOA increased the risk of dislocation (OR=0.892, CI=0.812 to 0.979, p=0.016) and thigh pain (OR=0.920, CI=0.851 to 0.995, p=0.037). CONCLUSIONS: The study demonstrates the relationship between FNOA and short-term radiological and clinical outcomes of patients after THA using a Tri-Lock femoral prosthesis. Inappropriate FNOA was significantly associated with failure of hip anatomical reconstruction and a higher risk of complications.

Top-cited works about exercise for knee osteoarthritis: a bibliometric analysis from 2000 to 2021.

OBJECTIVE: Previous trials demonstrated the effectiveness of exercise in improving pain and functional impairment in patients with knee osteoarthritis (KOA). However, a bibliometric analysis of top-cited papers on exercise treatment for KOA has not yet been conducted. The aim of the present study was to critically analyze the bibliometric characteristics of the most frequently cited articles on exercise treatment for KOA. MATERIALS AND METHODS: Publications about exercise treatment for KOA from 2000 to 2021 were searched from the Web of Science database. Two authors independently collected 100 top-cited articles, and a consensus was reached to form the final list. The title, journal, author, year of publication, country and institution of origin, total citations, citations in 2021, main topics, research nature, and level of evidence were extracted, and the publication trends in exercise treatment for KOA were evaluated. RESULTS: A total of 1,258 papers were retrieved from the database. According to the final list, clinical research accounted for 81% of the studies, but no statistical difference in the number of citations was found among the four types of articles (p=0.194). Seventy articles had a level of evidence of Ib, and no statistical differences in citations were found per level of evidence (p=0.767). Most of the top-cited articles were published between 2005-2014, and Dr Messier was the prominent writer in this field. CONCLUSIONS: This bibliometric study is the first to identify the most cited papers in exercise treatment for KOA research. Traditional Chinese exercise, comorbidity, and exercise adherence may be the next popular research trends that will receive more attention in the future.

[Retrospective analysis of short-term curative effect of total laparoscopic loop ileostomy reversal after radical resection of rectal cancer].

Objective: To investigate the safety, feasibility and short-term efficacy of total laparoscopic loop ileostomy reversal in patients after resection of rectal cancer. Methods: The clinical data of 20 patients who underwent total laparoscopic loop ileoscopic loop ileostomy after radical resection of rectal cancer at Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, or Beijing Chaoyang District Sanhuan Cancer Hospital from October 2019 to June 2020 were collected and retrospectively analyzed. Results: All patients had successfully underwent total laparoscopic ileostomy reversal without conversion to open surgery or discontinued operation. No perioperative related death cases were found. In the whole group, the median operation time was 97 (60-145) minutes and the median intraoperative blood loss was 20 (10-100) milliliters. The median Visual Analogue Scale (VAS) score was 1.9 (1-5) one day after the operation. Nobody needed to use additional analgesic drugs. The median time to grand activities was 25 (16-42) hours, the median time to flatus was 44 (19-51) hours, and the median hospitalization after operation was 6.9 (5-9) days. No patients underwent operation related complications such as operative incision infection, abdominal and pelvic infection, intestinal obstruction, anastomotic leakage, bleeding and so on. Conclusions: Total laparoscopic loop ileostomy reversal appears to be safe, feasible and with promising efficacy for selected patients.

Effects of Glycyrrhiza polysaccharide in diet on growth performance, serum antioxidant capacity, and biochemistry of broilers.

In the present study, we analyzed the effects of Glycyrrhiza polysaccharide (GCP) on growth performance, serum antioxidant capacity, and biochemistry of broilers. A total of 600, one-day-old AA broilers randomly divided into 5 treatment groups with 6 replicate pens of 20 birds per cage received dietary supplementation with GCP (0, 200, 500, 1,000, and 1,500 mg/kg) for 42 d. The supplementation of GCP linearly decreased (P < 0.05) feed conversion rate on day 22 to 42. Dietary supplementation with GCP reduced (P < 0.05) serum total cholesterol on day 21 and 42 and linearly improved (P < 0.05) albumin and high-density lipoprotein cholesterol. Dietary supplementation with 1,000 or 1,500 mg/kg GCP significantly increased (P < 0.05) serum total superoxide dismutase (T-SOD) activity on day 21 and 42 and reduced (P < 0.05) serum malondialdehyde content on 21 d. Dietary supplementation with 1,000 or 1,500 mg/kg GCP significantly improved (P < 0.05) interleukin-1ß (IL-1ß) and interferon-γ (IFN-γ) expressions in liver on day 21 and 42. At the end of the experiment, we randomly selected 20 broilers from 3 treatment groups (0, 1,000, and 1,500 mg/kg), respectively, to perform an lipopolysaccharide (LPS)-induced acute stress experiment. The 60 broilers were divided into 6 treatment groups with 10 birds per cage. The experiment was designed as a 3 × 2 factorial arrangement with GCP (0, 1,000, or 1,500 mg/kg) and LPS (injection of saline or 1 mg/kg body weight) levels as treatments. When the grouping was finished, the broilers were immediately intraperitoneally injected with LPS or normal saline. Six hours after challenged, serum antioxidant and liver immunity were analyzed. The results showed that dietary GCP prevented LPS-induced reductions in T-SOD activity and increases in malonaldehyde content (P < 0.05). Also, dietary GCP supplementation mitigated the LPS-induced increase in IL-1ß and IFN-γ in the liver. Supplementation with 1,500 mg/kg GCP showed the most optimal effect in broilers. GCP has the potential to be used as feed additive in broilers.

Front-face fluorescence spectroscopy combined with chemometrics to detect high proteinaceous matter in milk and whey ultrafiltration permeate.

Proteinaceous matter can leak into the permeate stream during ultrafiltration (UF) of milk and whey and lead to financial losses. Although manufacturers can measure protein content in the finished permeate powders, there is currently no rapid monitoring tool during UF to identify protein leak. This study applied front-face fluorescence spectroscopy (FFFS) and chemometrics to identify the fluorophore of interest associated with the protein leak, develop predictive models to quantify true protein content, and classify the types of protein leak in permeate streams. Crude protein (CP), nonprotein nitrogen (NPN), true protein (TP), tryptone-equivalent peptide (TEP), α-lactalbumin (α-LA), and ß-lactoglobulin (ß-LG) contents were measured for 37 lots of whey permeate and 29 lots of milk permeate from commercial manufacturers. Whey permeate contained more TEP than did milk permeate, whereas milk permeate contained more α-LA and ß-LG than did whey permeate. The types of protein leak were thus identified for predictive model development. Based on excitation-emission matrix (EEM) of high- and low-TP permeates, tryptophan excitation spectra were collected for predictive model development, measuring TP content in permeate. With external validation, a useful model for quality control purposes was developed, with a root mean square error of prediction of 0.22% (dry basis) and a residual prediction deviation of 2.8. Moreover, classification models were developed using partial least square discriminant analysis. These classification methods can detect high TP level, high TEP level, and presence of α-LA or ß-LG with 83.3%, 84.8%, and 98.5% cross-validated accuracy, respectively. This method showed that FFFS and chemometrics can rapidly detect protein leaks and identify the types of protein leak in UF permeate. Implementation of this method in UF processing plants can reduce financial loss from protein leaks and maintain high-quality permeate production.

[Association between fatty liver and type 2 diabetes in the baseline population of Jinchang Cohort].

Objective: To explore the association between fatty liver and type 2 diabetes mellitus (T2DM) in the baseline-population of Jinchang cohort study. Methods: Data from all the participants involved in the baseline-population of Jinchang cohort study was used, to compare the risks of T2DM in fatty liver and non fatty liver groups and to explore the interaction between family history or fatty liver of diabetes and the prevalence of T2DM. Results: Among all the 46 861 participants, 10 574 were diagnosed as having fatty liver (22.56%), with the standardized rate as 20.66%. Another 3 818 participants were diagnosed as having T2DM (8.15%) with standardized rate as 6.90%. The prevalence of T2DM increased in parallel with the increase of age (trend χ(2)=2 833.671, trend P<0.001). The prevalence of T2DM in the fatty liver group was significantly higher than that in the non-fatty liver group, both in men or women and in the overall population. Compared with the group of non-fatty liver, the risks of T2DM in fatty liver group were seen 1.78 times higher in males, 2.33 times in women and 2.10 times in the overall population, after adjustment for factors as age, levels of education, smoking, drinking, physical exercise, BMI, family history of diabetes and some metabolic indicators (pressure, TC, TG, uric acid, ALT, AST, gamma-glutamyl transferase). Date from the interaction model showed that fatty liver and family history of diabetes present a positive additive interaction on T2DM (RERI=1.18, 95%CI: 0.59-1.78; AP=0.24, 95%CI: 0.14-0.34; S=1.43, 95%CI: 1.21-1.69). Conclusions: Fatty liver could significantly increase the risk of T2DM and a positive additive interaction was also observed between fatty liver and family history of diabetes on T2DM. It was important to strengthen the prevention program on T2DM, in order to effectively control the development of fatty liver.

Dl-3-n-butylphthalide protects the blood brain barrier of cerebral infarction by activating the Nrf-2/HO-1 signaling pathway in mice.

OBJECTIVE: The aim of this study was to explore whether Dl-3-n-butylphthalide (DBT) could protect blood-brain barrier (BBB) of mice with experimental cerebral infarction and the relevant mechanism. MATERIALS AND METHODS: Adult male CD-1 mice were selected as the study objects. The permanent middle cerebral artery occlusion (MCAO) model was prepared by Longa's modified suture-occluded method. The mice were randomly divided into 3 groups: the sham operation group (Sham group), the cerebral infarction model group (CI group) and the DBT (120 mg/kg) intervention group (DBT group). Neurologic function deficits were evaluated by Longa's modified scoring method after 24 h of permanent MCAO. The wet and dry weight method was used for measuring water content in brain tissues. 2% 2,3,5-triphenyltetrazolium chloride (TTC) staining method was applied to determine the volume of cerebral infarction. Changes in the protein and messenger ribonucleic acid (mRNA) expression levels of matrix metallopeptidase 9 (MMP-9), claudin-5, vascular endothelial growth factor (VEGF), glial fibrillary acidic protein (GFAP), NF-E2 related factor 2 (Nrf-2) and heme oxygenase 1 (HO-1) in ischemic brain tissues were detected using immunohistochemistry, Western blotting and quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR). Ultrastructure changes in BBBs were observed under an electron microscope. RESULTS: DBT improved the neurologic function deficits of mice and reduced the infarction volume of mice with cerebral infarction. DBT alleviated edema and decreased the permeability of BBBs of mice with cerebral infarction. DBT down-regulated the expression of MMP-9 and up-regulated the expression of claudin-5 in brain tissues of mice with cerebral infarction. DBT increased the expressions of VEGF and GFAP. DBT improved the ultrastructure in capillary endothelial cells of BBBs and increased the expressions of Nrf-2 and HO-1. CONCLUSIONS: DBT may protect BBB by activating the Nrf-2/HO-1 signaling pathway, thus achieving its protective effect on the brain.

Large-scale screening and characterisation of Lemna aequinoctialis and Spirodela polyrhiza strains for starch production.

Duckweed is considered a promising feedstock for bioethanol production due to its high biomass and starch production. Selection of duckweed strains with high starch accumulation is essential for application of duckweeds to bioethanol production. Geographic differentiation had a large influence on genetic diversity of duckweeds. Biomass production, starch content and starch amount in geographically isolated strains of 20 Lemna aequinoctialis and Spirodela polyrhiza were calculated to evaluate their potential for bioethanol production. The influence of different collection time, culture medium and NaCl concentration on starch accumulation of the best strains were analysed. The results showed that biomass production, starch content and starch production of duckweeds demonstrated clonal dependency. The best strain was L. aequinoctialis 6000, with biomass production of 15.38 ± 1.47 g m-2 , starch content of 28.68 ± 1.10% and starch production of 4.39 ± 0.25 g m-2 . Furthermore, starch content of L. aequinoctialis 6000 was highest after 8 h of light, tap water was the best medium for starch induction, and NaCl did not induce starch accumulation. This study suggests duckweed biomass production and starch production demonstrate clonal dependency, indicating that extensive clonal comparisons will be required to identify the most suitable isolates for duckweed selective breeding for bioethanol.

[The Value of SMI in the Evaluation of interventional therapy of liver cancer].

Objective: To evaluate the value of super microvascular imaging(SMI) for evaluating the effect of interventional therapy of liver cancer. Methods: A total of 30 patients with 40 leisions were enrolled in this study, from the tumor intervention department in the third affiliated hospital of suzhou university.This patients were underwent TACE, after the treatment 1 month, CDFI, SMI, and CT were study respectively. Using the continuity correction McNemar matching chi-square test, with P < 0.05 for the standard , CDFI and SMI shows the difference in monitoring the microvascular imaging in and around the tumors leisions. Results: A total of 30 patients, 12 cases were primary liver cancer (7 cases combined with liver cirrhosis), 18cases were metastatic liver cancer; 30 cases including 25 single and 5 multiple. Significant difference were found between CT and SMI in detecting blood flows inside the lesion (χ² = 8.642 9, P < 0.05), and were also found between CT and CDFI in detecting blood flows inside the lesion (χ² = 16.961 5, P < 0.05). The AUROC, sensitivity, specificity, accuracy, PPV and NPV of CDFI were 0.647, 29.4%、100%、29.4%、100%、20.0%, while in SMI were 0.809, 61.8%、100%、61.7%、100%、31.5%, respectively. Conclusions: SMI SMI can detect the microvascular inside the lesions. This new method was superior to CDFI, achieving the same effect as CT.

MicroRNA-32 functions as a tumor suppressor and directly targets EZH2 in uveal melanoma.

MicroRNA-32 (miR-32) has been shown to be dysregulated in some human malignancies and this has been found to be correlated with tumor progression. However, its role in uveal melanoma formation and progression remains largely unknown. Thus, the aim of this study was to explore the expression and function of miR-32 in human uveal melanoma. Using quantitative reverse transcription-polymerase chain reaction, we detected miR-32 expression in uveal melanoma tumor tissues and cell lines. The effects of miR-32 on the biological behavior of uveal melanoma cells were also investigated. Finally, the potential regulatory function of miR-32 on EZH2 expression was confirmed. miR-32 expression levels were significantly downregulated in uveal melanoma samples and cell lines (both P < 0.01). Ectopic expression of miR-32 could inhibit uveal melanoma cell proliferation, migration, and invasion, and promote cell apoptosis in vitro. Further, EZH2 was confirmed as a direct target of miR-32 by using the luciferase reporter assay. These findings indicate that miR-32 may function as a novel tumor suppressor in uveal melanoma and could be a potential therapeutic target for this disease.

Correlation of miR-494 expression with tumor progression and patient survival in pancreatic cancer.

MicroRNA-494 (miR-494) expression is aberrant in various types of human cancer. However, the prognostic value of miR-494 in pancreatic cancer remains unclear. The level of miR-494 expression was determined in 99 pairs of primary pancreatic cancer and their corresponding, adjacent non-tumor tissues by using quantitative reverse transcriptase polymerase chain reaction. We also analyzed the associations between miR-494 expression and clinicopathological features. The survival correlations were analyzed by using the Kaplan-Meier method and Cox proportional hazards model. The level of miR-494 expression was significantly downregulated in pancreatic cancer tissues (mean relative expression level ± SD, 0.48 ± 0.11) as compared to matched adjacent normal tissues (1.80 ± 0.28, P < 0.05). We found significant correlations between the miR-494 expression levels and TNM stage (P = 0.009), lymphatic invasion (P = 0.036), vascular invasion (P = 0.011), distant metastasis (P = 0.007), and tumor grade (P = 0.031). Pancreatic cancer patients with a low miR-494 expression level had a shorter overall survival than those with a high miR-494 expression level (P < 0.05). Reduced miR-494 expression in pancreatic cancer tissues is correlated with tumor progression and might be an independent, poor prognostic factor for patients with pancreatic cancer.

Nucleotide mismatches of foot-and-mouth disease virus during replication.

As there is a lack of error correction mechanisms during RNA replication, foot-and-mouth disease virus (FMDV) has a very high mismatch rate, which leads to a high mutation rate, in the range of 10(-3) to 10(-5) per nucleotide site per genome replication. We examined the nucleotide mismatch of FMDV during replication, based on the whole genomes of the 7 serotypes retrieved from NCBI. With the Mega bio-software, SPSS, and Microsoft Excel, we studied the nucleotide differences compared to the sequence in the RefSeq database, and developed two probable mutation models, i.e., once mutation model and complication mutation model. Further analysis on the nucleotide mismatch during replication was made. We found that FMDV share similar difference rates between nucleotide and reverse differences, for example the mutation U→C and C→U. We also found that each nucleotide has its domain mismatch, and the virus kept a constant nucleotide composition during mutations.

A cross-sectional study of sputum handling by and supervision of patients with pulmonary tuberculosis treated at home in China.

Disposal of sputum from patients with pulmonary tuberculosis (TB) who are treated at home is an important aspect of preventing the spread of TB. However, few studies have examined disposal of sputum by patients with TB who are treated at home. Patients with pulmonary TB who are treated at home were surveyed regarding sputum handling and supervision. A cross-sectional survey of a representative sample of patients with pulmonary TB who are treated at home was conducted in Shandong Province. Participants were individuals with TB who had been registered with a local agency responsible for TB control. Participants completed a questionnaire with both qualitative and quantitative questions. How sputum was handled was determined and factors associated with sputum disposal were analyzed using a non-parametric test, logistic regression, and content analysis. Responses were received from 720 participants. Patients expectorated sputum 4.56 ± 10.367 times a day, and 68.6% of patients responded that they correctly disposed of their sputum. Supervision as part of TB control focused on the efforts of health agencies and paid little attention to waste management by patients. A non-parametric test showed that sputum disposal was significantly associated with gender, age, education, sputum smear results, attitudes toward waste management, and attitudes toward supervision (all p < 0.05). Logistic regression analysis showed that gender (OR = 0.482, 95% CI: 0.329-0.704), sputum smear results (OR = 1.300, 95% CI: 1.037-1.629), and level of education (OR = 0.685, 95% CI: 0.528-0.889) were associated with receipt of TB health education (all p < 0.05). Sputum handling by and supervision of patients with pulmonary TB who are treated at home is severely wanting. From a policy perspective, special attention should be given to the definition, details, and methods of supervision of waste management by patients with TB to give them relevant health education and enhance their willingness to be supervised. A financial incentive should be provided to health workers supervising management of TB-related waste.

Endothelial nitric oxide synthase gene polymorphisms and essential hypertension in Han Chinese.

We examined the effect of polymorphisms in the endothelial nitric oxide synthase gene on the risk for essential hypertension in a Han Chinese population through a meta-analysis of data from 15 studies. Associations between increased risk for essential hypertension and 4b/a were obtained in a dominant model and allele contrast (aa + ab vs bb: odds ratio (OR)(FE) = 1.26, 95% confidence interval (CI) = 1.10-1.44; a vs b allele: OR(FE) = 1.23, 95%CI: 1.09-1.40). Four studies with sample sizes over 500 produced similar results. No evidence of publication bias was found. Also, no significant heterogeneity was observed among these studies. When we examined the G894T polymorphism, we found a marginally significant association for allele contrast and the recessive model when all the eligible studies were pooled together. However, there was no evidence for a significant association after the exclusion of two studies deviating from Hardy-Weinberg equilibrium in the control group. Heterogeneity among studies was observed. Results of cumulative and recursive cumulative meta-analysis indicated that more studies are needed to objectively determine the effects of these two polymorphisms.

The effect of grain size of rock phosphate amendment on metal immobilization in contaminated soils.

When rock phosphates (RP) are used to remediate Pb-contaminated soils, their effectiveness is likely affected by their grain size. In this study, the effect of grain size of rock phosphate on the effectiveness of heavy metal immobilization in two contaminated soils was measured in pot experiment. Rock phosphate was used with four different grain sizes: <35, 35-72, 72-133 and 133-266microm. The application rate of rock phosphate in two soils was determined based on P/metals (Pb, Zn, Cu and Cd) molar ratio of 5.0 in the soils. The results showed that rock phosphate of the smallest grain size (<35microm) was superior to all of other grain sizes more than 35microm for reducing uptake in plant (Brassica oleracea L.) shoots for Cd (19.6-50.0%), Pb (21.9-51.4%) and Zn (22.4-34.6%), respectively, as compared with the soil without application of rock phosphate. Sequential extraction analysis indicated that rock phosphate was most effective for soil Pb to induced transformation from non-residual fractions to a residual fraction than that for Zn and Cd. Such transformation was probably through dissolution of Pb associated with exchangeable (EX), organic fraction (OC), acidic fraction (AC) and amorphous Fe and Al oxides-bound (OX) fraction and precipitation of pyromorphite-like minerals. Results suggested that the rock phosphate with small grain size was superior to that with large grain size for in situ remediation technology.

Sesquiterpenoids from Magnolia grandiflora.

Three new sesquiterpenoids were obtained from the leaves of Magnolia grandiflora L. Their structures were determined as 6 alpha,11-dihydroxy-12,13-diacetoxyelem-1,3-diene, 4 alpha,6 alpha,10 alpha-trihydroxy-13-acetoxyguaia-11-ene, and 12,13-diacetoxyguaia-4 alpha,6 alpha,10 alpha,11-tetraol on the basis of spectral evidence. In addition, the known sesquiterpenoid magnograndiolide was also obtained.

Two new germacranolides from Magnolia grandiflora.

Two new sesquiterpenoids, 4,5-epoxy-13-methoxy-1(10)-germacren-12,6-olide and 4,5-epoxy-13-acetoxy-1(10)-germacren-12,6-olide, were isolated from the leaves of Magnolia grandiflora, together with six known compounds, 2alpha-hydroxy-dihydroparthenolide, parthenolide, costunolide, syringaresinol, (+) medioresinol and 6,7-dimethoxycoumarin. The structures of the new compounds were elucidated on the basis of spectroscopic methods and X-ray diffraction.

ent-Pimarane derivatives from Dysoxylum hainanense.

Four ent-pimarene diterpenoids. ent-18-acetoxy-8(14)-pimarene-15S, 16-diol, ent-18-acetoxy-16-hydroxy-8(14)-pimaren-15-one, ent-16,18-dihydroxy-8(14)-pimaren-15-one and ent-19-nor-4,16,18-trihydroxy-8(14)-pimaren-15-one, together with three known damarane triterpenoids, richenoic acid, eichleriainic acid and shoreic acid were isolated from the bark of Dysoxyhum hainanense Merr. Their structures were elucidated on the basis of spectroscopic techniques. The absolute configurations of four diterpenoids were assigned as ent-pimarene type by chemical transformation and by co-occurrence in the plant as well as by negative optical rotations for four compounds.

[Propagation of hepatitis E virus in several cell lines including human embryo lung diploid cell KMB17].

OBJECTIVE: To investigate the hepatitis E virus (HEV) sensitive cells and its tissue culture conditions. METHODS: The HEV from dejecta supernatant of patients with acute hepatitis E was amplified and activated by passaged in Rhesus. Then, the positive dejecta samples of infected monkeys were dealt with super-centrifugation and virus for culture was obtained. Various human-derived (including KMB17, A549, BEL7402, and Hela) and non-human primates derived cells (Vero) were inoculated with HEV. Sensitivity of cells to HEV was measured by CPE (cytopathic effect), RT-PCR and immunofluorescence. RESULTS: CPE in KMB17, A549 and BEL7402 cells appeared during 7-9 days, meanwhile, cells shelled during 11-13 days on the first filial generation. The existence of HEV genome +RNA and replicated -RNA was still detectable by RT-PCR after the tenth filial generation. Neither CPE nor amplification of HEV genome RNA could be detected in Hela and Vero cells after the second to fourth filial generation. HEV could also be detected from inoculated KMB17 cells by immunofluorescence and RT-PCR. CONCLUSIONS: It indicates that KMB17, A549 and BEL7402 cells are sensitive to HEV under the experimental culture conditions, while Hela and Vero cells are insensitive. Tissue culture system of HEV in certain filial generation is established.