The chemical constituents of Taxus chinensis var. mairei cell cultures were investigated by chromatographic methods, including silica gel column chromatography, Sephadex LH-20 and preparative HPLC. Thirteen compounds were isolated from the 80% ethanol extract of cultured cells and their structures were elucidated by spectral data and physicochemical properties, which were identified as 2α,4α,7β,9α,10β-pentaacetoxy-14β-hydroxytax-11-ene (1), 2α,4α,7β,9α,10β-pentaacetoxytax-11-ene (2), 1β-deoxybaccatin VI (3), 2α-acetoxytaxusin (4), taxuyunnanine C (5), yunnanxane (6), 2α,5α,10β-triacetoxy-14β-propionyloxy-4 (20), 11-taxadiene (7), 2α,5α,10β-triacetoxy-14β-isobutyryloxy-4 (20), 11-taxadiene (8), 2α,5α,10β-triacetoxy-14β-(2'-methyl)butyryloxy-4 (20), 11-taxadiene (9), 13-dehydroxylbaccatin III (10), 13-dehydroxy-10-deacetylbaccatin III (11), paclitaxel (12) and (13) β-sitosterol. Among them, compound 1 is a new compound, and compounds 2, 4, 10 and 11 are isolated from the cell culture of Taxus chinensis var. mairei for the first time.
Alkenes ; analysis ; Cell Culture Techniques ; Cells, Cultured ; Diterpenes ; analysis ; Molecular Structure ; Paclitaxel ; analysis ; Sitosterols ; analysis ; Taxoids ; analysis ; Taxus ; chemistry

OBJECTIVETo investigate the effect of potassium iodide on the expression of nuclear factor-kappaB and fibronectin.

METHODSThe experiment was performed with 72 SD rats weighing about 180-220 g. The animals were randomly assigned into nine groups. Group A, B, C (n=8) served as control and were fed with distilled water for 1 month, 2 month, 3 month respectively. Group D, E, F (n=8) served as lead exposed and were fed with water with 0.5% lead acetate for 1 month, 2 month, 3 month respectively. Group G, H, I (n=8) served as potassium iodide and lead exposed and were treated with 0.5% lead acetate simultaneously taking potassium iodide 3 mg/100 g weight by intragastric administration for 1 month, 2 month, 3 month respectively. Animals of different groups were sacrificed at the end of the treatment. Ultrastructure of kidney was observed by electron microscopy; Expression of NF-kappaB and FN protein and mRNA in kidney were measured respectively by immunohistochemistry and RT-PCR.

RESULTSElectron microscopic examination revealed potassium iodide could restrain the denaturalization in epithelial cells and mitochondrial cristae. The expressions of NF-kappaB protein (0.2315 +/- 0.0624, 0.3213 +/- 0.0740, 0.4729 +/- 0.0839) and mRNA (0.4370 +/- 0.0841, 0.5465 +/- 0.0503, 0.6443 +/- 0.0538) in all the lead exposed groups continuously increased compared with correspondent control groups; Group I was decreased obviously compared with group F. The expressions of FN protein (0.4243 +/- 0.0595, 0.4917 +/- 0.0891) and mRNA (0.8650 +/- 0.0880, 0.8714 +/- 0.0980) in group E and F increased compared with group B and C, but the expressions of FN protein in group I significantly decreased compared with group F; The expressions of FN mRNA in Group H and I significantly decreased compared with group E and F.

CONCLUSIONThe potassium iodide can ameliorate renal ultrastructure and degrade expression of nuclear factor-kappaB and fibronectin induced by lead.

Animals ; Disease Models, Animal ; Fibronectins ; genetics ; metabolism ; Kidney ; drug effects ; metabolism ; ultrastructure ; Kidney Diseases ; chemically induced ; metabolism ; pathology ; Lead Poisoning ; complications ; metabolism ; pathology ; Male ; NF-kappa B ; genetics ; metabolism ; Potassium Iodide ; pharmacology ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley

Hemophilia B (HB) is a recessive X-linked inherited disorder, the pathogenesis of HB is deficiency or functional abnormalities of coagulation factor IX, which is caused by F9 gene mutations. To explore the mechanism of its molecular pathology, 40 patients with HB were studied with polymerase chain reaction (PCR) and direct sequencing. The diagnosis of HB patients were based on clinical manifestation and deficient factor IX activity in plasma. DNA was routinely extracted from peripheral blood cells of the patients and their relatives, all the 8 exons and their flanking boundaries were amplified by PCR, and the PCR products were screened by direct sequencing. Mutations which were found in study need to exclude polymorphism. The results showed that 34 mutations were confirmed in 40 HB patients, including 6 nonsense mutations, 24 missense mutations, 2 splice site mutations and 2 frame mutations for 1 or 2 nucleotide insertion. After retrieved, 4 missense mutations and 1 frameshift mutation were found for the first time. Among the 34 mutations, 2 mutations in signal peptide, 7 mutations in propeptide and gla domain, 7 mutations in epidermal growth factor-like domain, 3 mutations in activation domain, 15 mutations in serine protease or catalytic domain. It is concluded that gene analysis can directly explain molecular mechanism of hemophilia B and also provides the foundation for further studies to the function of coagulation factor IX. There is obvious heterogeneity in F9 gene mutation and missense mutation is still the main way of mutation, which are closely related to clinical features. DNA sequencing and linkage analysis are efficient methods for HB carriers and prenatal gene diagnosis.
Base Sequence ; DNA Mutational Analysis ; Factor IX ; genetics ; Hemophilia B ; diagnosis ; genetics ; Humans ; Male

OBJECTIVESTo analyze retrospectively the formation and histological changes of sclerosis rim in patients with osteonecrosis of the femoral head (ONFH), and to study the relationship between bone morphogenetic proteins (BMP4) and sclerosis rim, so as to acquire experimental and theoretical basis on individualized treatment for ONFH patients.

METHODSFrom November 2005 to November 2007, 184 hips of steroid-induced ONFH inpatients were collected. The mean age was (47 ± 7) years, the patients were divided into high (more than 54 years old), middle (40 - 54 years old) and low (less than 40 years old) age groups. Their clinical data were analyzed retrospectively according to gender and age. Parts of the femoral heads were selected for the study, including 18 hips in high age group, 11 hips in low age group and 20 hips in middle age group. Each 10 hips were selected with or without sclerosis rim. The femoral heads were cut along middle coronal plane, their weight-bearing and non-weight-bearing areas were used for the study. The specimens were processed by routine HE staining and picric acid-Sirius red staining and electron microscopy preparation and immunohistochemistry stain. The average optical density of BMP4 protein was calculated by image analysis software.

RESULTSThe trabecular of sclerosis rim was thickening and disorder. But its osteocytes were normal and with high secretion. The ratio of sclerosis rim was 71.4% (105/147) in middle age ONFH patients, which was significantly higher than the low age group patients (45.5%, 5/11) and high age group patients (38.5%, 10/26) (P < 0.01). The optical density of BMP4 in middle age ONFH patients was 0.32 ± 0.14, which was significantly higher than the low age group 0.20 ± 0.17 and high age patients 0.19 ± 0.27 (P < 0.05). The optical density was 0.16 ± 0.11 in ONFH patients without sclerosis rim, which was significantly lower than with sclerosis rim (0.28 ± 0.13) (P < 0.01). The time from hip pain to joint replacement in patients with sclerosis rim was (49 ± 11) months, and (15 ± 2) months without sclerosis rim. There was significant difference between the two groups (P < 0.01).

CONCLUSIONSThe formation of sclerosis rim is positively related to the expression of BMP4, and high expression of BMP maybe promote the formation of sclerosis rim.

Adult ; Bone Morphogenetic Protein 4 ; metabolism ; Female ; Femur Head ; metabolism ; pathology ; Femur Head Necrosis ; metabolism ; pathology ; Humans ; Male ; Middle Aged ; Retrospective Studies

OBJECTIVETo construct pseudovirus bearing H5N1 HA based on a lentivirus vector system. Then we study the biological feature of the pseudovirus. With the newly established viral particles, we performed the serological tests.

METHODSH5N1 avian influenza virus that isolated from human case was cloned to construct pLP-HA, then pLP-HA co-transfected with lentivirus vector plasmids pLP1, pLP2 and pEmGFP into 293T cells. The supernatant was harvested 48h post-transfection. Concentrated by super centrifuge, the pseudotyped viruses were analyzed by infection test, HA test and micro-neutralization test. At the same time, optimized HA gene and a Vietnam H5N1 HA gene were used to construct pseudotyped virus for comparison.

RESULTSPseudotyped virus particles can be observed with electronic microscope. Western-blot revealed that HA glycoprotein can be expressed in the virions. Our neutralization assay by using the pseudoviruses was comparable with the conventional microneutralization assay with wild-type viruses. A high degree of correlation was detected.

CONCLUSIONPseudotyped Viruses coated with HA of H5N1 High Pathogenic Avian Influenza were successfully constructed; it can be used to for the microneutralization assay. The HA gene from different sources affect the efficiency of the packaging of the pseudovirus. But the optimized HA gene can not obviously improve packaging efficiency of the pseudovirus.

Animals ; Cell Line ; Dogs ; Gene Expression ; Genetic Engineering ; Genetic Vectors ; genetics ; metabolism ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; metabolism ; Humans ; Influenza A Virus, H5N1 Subtype ; genetics ; physiology ; ultrastructure ; Influenza, Human ; virology ; Lentivirus ; genetics ; metabolism ; Virion ; genetics ; physiology ; ultrastructure

BACKGROUNDSystemic administration of bisphosphonates has shown promising results in the treatment of osteonecrosis of the femoral head (ONFH). However, few studies have evaluated the efficacy of local zoledronate (ZOL) administration in the treatment of ONFH. The purpose of this study was to investigate whether local administration of bisphosphonate-soaked hydroxyapatite (HA) could improve bone healing in an experimental rabbit model of ONFH.

METHODSThis experimental study was conducted between October 2014 and June 2015. Forty-five rabbits underwent simulated ONFH surgery. Immediately following surgery, they were divided into three groups: model (untreated, n = 15), HA (treated with HA alone, n = 15), and HA + ZOL (treated with HA soaked in a low-dose ZOL solution, n = 15). Histological, immunohistochemical, and quantitative analyses were performed to evaluate bone formation and resorption 2, 4, and 8 weeks after surgery.

RESULTSGross bone matrix and hematopoietic tissue formation were observed in the HA + ZOL group 4 weeks after surgery. The immunohistochemical staining intensities for 5-bromodeoxyuridine, runt-related transcription factor 2, osteocalcin, osteopontin, and osteoprotegerin were significantly higher in the HA + ZOL group than that in the model (P < 0.001, P< 0.001, P< 0.001, P< 0.001, and P = 0.018, respectively) and HA groups (P = 0.003, P = 0.049, P< 0.001, P = 0.020, and P = 0.019, respectively), whereas receptor activator of the nuclear factor-κB ligand staining intensity was significantly lower in the HA + ZOL group than that in the model and HA groups (P = 0.029 and P = 0.015, respectively) 4 weeks after surgery. No significant differences in bone formation or bone resorption marker expression were found between the three groups 2 or 8 weeks after surgery (P > 0.05).

CONCLUSIONSLocal administration of HA soaked in a low-dose ZOL solution increased new bone formation while inhibiting bone resorption in an animal model of ONFH, which might provide new evidence for joint-preserving surgery in the treatment of ONFH.

Animals ; Diphosphonates ; administration & dosage ; therapeutic use ; Durapatite ; administration & dosage ; therapeutic use ; Female ; Femur Head Necrosis ; drug therapy ; metabolism ; Imidazoles ; administration & dosage ; therapeutic use ; Immunohistochemistry ; Male

Objective:To evaluate the effect of canine-assisted therapy on family function of autism spectrum disorder. Methods:The randomized controlled trials (RCT) about the effect of canine-assisted therapy on family function of autism spectrum disorder were retrieved from the Cochrane Library, PubMed, Web of Science, CNKI, CBM, VIP, and Wanfang Data from establishment to February, 2021. Brief Family Assessment Measure-III-General Scale (FAM-III-GS) was used to access the family function, and Children's Depression Inventory-Second Edition (CDI-2) and Spence Children's Anxiety Scale (SCAS) were used to access the psychological behavior. The quality of included studies was evaluated according to the method recommended by the Cochrane Collaboration and Joanna Briggs Institute Reviewers' manual. RevMan 5.4 software was used for meta-analysis. Results:Finally, four RCTs involving 190 patients were included. There were too less RCTs to do a meta-analysis. Two RCTs showed that the score of FAM-III-GS improved in the experimental group than in the control group. Conclusion:Canine-assisted therapy might improve the family function of patients with autism spectrum disorder.

BACKGROUNDHigh intraocular pressure (IOP) and low central corneal thickness (CCT) are important validated risk factors for glaucoma, and some studies also have suggested that eyes with more deformable corneas may be in higher risk of the development and worsening of glaucoma. In the present study, we aimed to evaluate the association between corneal biomechanical parameters and asymmetric visual field (VF) damage using a Corvis-ST device in patients with untreated normal tension glaucoma (NTG).

METHODSIn this observational, cross-sectional study, 44 newly diagnosed NTG patients were enrolled. Of these, 31 had asymmetric VF damage, which was defined as a 5-point difference between the eyes according to the Advanced Glaucoma Intervention Study scoring system. Corneal biomechanical parameters were obtained using a Corvis-ST device, such as time from start until the first and second applanation is reached (time A1 and time A2, respectively), cord length of the first and second applanation (length A1 and length A2, respectively), corneal speed during the first and second applanation (velocity A1 and velocity A2, respectively), time from start until highest concavity is reached (time HC), maximum amplitude at the apex of highest concavity (def ampl HC), distance between the two peaks at highest concavity (peak dist HC), and central concave curvature at its highest concavity (radius HC).

RESULTSTime A1 (7.19 ± 0.28 vs. 7.37 ± 0.41 ms, P = 0.010), length A1 (1.73 [1.70-1.76] vs. 1.78 [1.76-1.79] mm, P = 0.007), length A2 (1.58 [1.46-1.70] vs. 1.84 [1.76-1.92] mm, P< 0.001), peak dist HC (3.53 [3.08-4.00] vs. 4.33 [3.92-4.74] mm, P = 0.010), and radius HC (6.20 ± 0.69 vs. 6.59 ± 1.18 mm, P = 0.032) were significantly lower in the worse eyes than in the better eyes, whereas velocity A1 and def ampl HC were significantly higher (0.156 [0.149-0.163] vs. 0.145 [0.138-0.152] m/s, P = 0.002 and 1.19 ± 0.13 vs. 1.15 ± 0.13 mm, P = 0.005, respectively). There was no significant difference in time A2, velocity A2, and time HC between the two groups. In addition, no difference was observed in IOP, CCT, and axial length. In the univariate and multivariate analyses, some of the Corvis-ST parameters, including time A1 and def ampl HC, were correlated with known risk factors for glaucoma, and there was also a significant positive correlation between def ampl HC and age.

CONCLUSIONSThere were differences in dynamic corneal response parameters but not IOP or CCT between the paired eyes of NTG patients with asymmetric VF damage. We suggest that the shape of the cornea is more easily altered in the worse eyes of asymmetric NTG patients.

Aged ; Biomechanical Phenomena ; physiology ; Cornea ; metabolism ; physiology ; Cross-Sectional Studies ; Female ; Glaucoma ; metabolism ; physiopathology ; Humans ; Intraocular Pressure ; physiology ; Low Tension Glaucoma ; metabolism ; physiopathology ; Male ; Middle Aged ; Multivariate Analysis ; Prospective Studies ; Visual Fields ; physiology

Objective:To evaluate the effect of Kinesio taping on chronic nonspecific low back pain (CNLBP). Methods:The Cochrane Library, PubMed, Web of Science, CNKI, CBM, VIP, and Wanfang Data were searched for the randomized controlled trials (RCT) about the effect of Kinesio taping on CNLBP from establishment to January, 2019. The included studies were evaluated according to the method recommended by the Cochrane Collaboration. RevMan 5.3 software was used to analyze the extracted data. Results:Finally, nine RCTs involving 545 patients were included. Meta-analysis showed that the effect of Kinesio taping was significantly different in the improvement of pain compared with the non-stimulated group (MD = -0.76, 95%CI: -1.43 to -0.08, P = 0.03), the difference might be significant compared with the sham stimulation group (MD = -1.10, 95%CI: -2.22 to 0.02, P = 0.05); For improving dysfunction, the Oswestry Disability Index (ODI) scores were better in the Kinesio taping group than in the non-stimulation group (MD = -6.02, 95%CI: -8.63 to -3.41, P < 0.001) and the sham stimulation group (MD = -4.11, 95%CI: -5.82 to -2.41, P < 0.001), however, their was no significant difference in Roland Morris Disability Questionnaire (RMDQ) score between the Kinesio taping group and the non-stimulated group (MD = 0.69, 95%CI: -2.35 to 3.74, P = 0.66), and between the Kinesio taping group and the sham stimulation group (MD = -0.17, 95%CI: -1.43 to 1.08, P = 0.78). Conclusion:For the patients with CNLBP, the intervention of Kinesio taping could alleviate pain and improve function.

OBJECTIVE: Epstein-Barr virus associated gastric cancer (EBVaGC) is different from the traditional gastric cancer (Epstein-Barr virus non-associated gastric cancer, EBVnGC), and has unique clinicopathological features. This study investigated the largest single center cancer series so as to establish the clinicopathological and molecular characteristics of EBVaGC in China. METHODS: A retrospective analysis was conducted on EBVaGC and EBVnGC patients diagnosed at Peking University Cancer Hospital from 2003 to 2018 by comparing their clinicopathological features and prognosis. The gastric cancer (GC) dataset of public database was analyzed to obtain differentially expressed genes. The expression of important genes and their association with prognosis of GC were verified in GC tissues from our hospital. RESULTS: In this study, 3 241 GC patients were included, and a total of 163 EBVaGC (5.0%) patients were identified. Compared with EBVnGC, EBVaGC was higher in male and younger patients, and positively associated with remnant GC, poorly differentiated adenocarcinoma, and mixed type GC. EBVaGC was inversely related to lymph node metastasis. The 5-year survival rate of EBVnGC and EBVaGC was 59.6% and 63.2% respectively (P<0.05). In order to explore molecular features of EBVaGC, the Cancer Genome Atlas (TCGA) dataset was analyzed (n=240), and 7 404 significant differentially expressed genes were obtained, involving cell proliferation, apoptosis, invasion and metastasis. The down-regulated invasion/metastasis gene SALL4 and the up-regulated immune checkpoint gene PD-L1 were important molecular features of EBVaGC. Validation of these two genes in large GC series showed that the majority of the EBVaGC was SALL4 negative (1/92, 1.1%, lower than EBVnGC, 303/1 727, 17.5%), and that PD-L1 was mostly positive in EBVaGC (81/110, 73.6%, higher than EBVnGC, 649/2 350, 27.6%). GC patients with SALL4 negative and PD-L1 positive were often associated with better prognosis. CONCLUSION EBVaGC is a unique subtype of GC with less metastasis and a good prognosis. It also has a distinct molecular background. The down-regulation of invasion/metastasis gene SALL4 and up-regulation of immune checkpoint gene PD-L1 are important molecular features.
China ; Epstein-Barr Virus Infections/complications* ; Female ; Herpesvirus 4, Human ; Humans ; Male ; Retrospective Studies ; Stomach Neoplasms/etiology*