Objective: To study the chemical constituents from Scilla scilloides. Methods: The chemical constituents were isolated and purified by column chromatography and their structures were identified by spectral data. Results: Eighteen compounds were isolated from S. scilloides. Their structures were identified as scillascillin (1), 2-hydroxy-7-O-methylscillascillin (2), 4'-demethyleucomin (3), 5-hydroxy-7-methoxy-3-(4-hydroxybenzylidene) chroman-4-one (4), 4'-demethyl-3, 9-dihydroeucomin (5), 3'-hydroxy-3, 9-dihydroeucomin (6), 8-O-demethyl-7-O-methyl-3, 9-dihydropunctatin (7), apigenin (8), luteolin (9), chrysoeriol (10), 3-dehydro-15-deoxoeucosterol (11), 15-deoxoeucosterol (12), 4-allylpyrocatechol (13), norlichexanthone (14), drimiopsin C (15), 6-feruloylcatalpol (16), catalposide (17), and specioside (18). Conclusion: Iridoids (16-18) are isolated from the genus Scilla L. for the first time. Compounds 4, 7-10, and 13-18 are isolated from the plants of this genus for the first time.

AIM To speculate the hypoglycemic mechanism for rats with type 2 diabetes by exploring the therapeutic effects of leaf aqueous extract from Cyclocarya paliurus on liver insulin receptor (InsR) and insulin receptor substrate 2 (IRS-2).METHODS The diabetic rat model was established through intraperitoneal injection of streptozotocin and fed with high-fat diet.The moleled rats were equally assigned into the control group and leaf aqueous extract from Cyclocarya paliurus group (extract group).After the test extract was orally administrated for four weeks,body weight,urine output,food intake,water intake and fasting blood-glucose (FBG) were measured,and the levels of serum insulin,InsR and IRS-2 mRNA in liver tissue were investigated in rats.RESULTS Compared with the control group,the extract group showed a reduction in urine output,food intake,water intake,FBG and insulin levels.Meanwhile,the rats' body weights in extract group were presented a trend to increase.The gene expressions of InsR and IRS-2 in liver tissue were up-regulated.Moreover,the insulin sensitivity was improved.CONCLUSION The leaf aqueous extract from Cyclocarya paliurus can reduce FBS,improve insulin sensitivity,which may be associated with the increase of InsR and IRS-2 gene expression in liver tissue.

Case-Control Studies ; Creatine ; urine ; Female ; Humans ; Hypoxia-Ischemia, Brain ; diagnosis ; urine ; Infant, Newborn ; Lactic Acid ; urine ; Male ; Predictive Value of Tests

Objective To investigate the prevalence and pattern of androgenetic alopecia (AGA) in Shanghai through a community-based survey. Methods A cluster sampling survey was done among the residents in Beixinjing Community, Changning District, Shanghai. All the subjects were asked to fill a questionnaire to provide their general information, including sex, age, native place, physical status, life habit, family history, etc. The diagnosis of AGA was made by dermatologists. To determine the pattern of hair loss,Norwood-Hamilton classification system and Ludwig classification system were used for male AGA and female AGA, respectively. All the data were statistically analyzed by EpiData and SPSS11.5 software. Results Totally, 7056 subjects completed the questionnaire, including 3519 males and 3537 females, and the response rate was 72.5%. AGA was diagnosed in 809 patients, consisting of 701 males aging from 19 to 91 years (mean 64.16±11.9 years) and 108 females aging from 35 to 91 years (mean 70.46±18.89 years). The standardized prevalence (SP) was 9.47% in total, 15.73% in males and 2.73% in females; the difference was significant between males and females (χ2=356.00, P<0.001). A family history of AGA was observed in 52.7% of all subjects including 391 (55.78%) males and 35 (32.41%) females. Type Ⅲ vertex involvement was the most common type in men aging from 20 to 70 years old, and type Ⅵ in those over 70 years old. Grade Ⅰ and Ⅱ predominated in female AGA. Conclusions The results of this survey indicate that the prevalence of AGA is remarkably higher in men than that in women. Furthermore, the prevalence is steadily increased with advancing age in Shanghai.

OBJECTIVETo investigate the histomorphology and the expressions of the proliferation marker Ki-67 and estrogen receptor in the uterus of mice with autoimmune premature ovarian failure (POF) induced by zona pellucida 3 peptide (pZP3).

METHODSAutoimmune POP models were established in 20 female BALB/c mice (7-8 weeks old) by immunization with pZP3 and another 20 mice served as the control group. The POP models were verified by vaginal cytology, serum sex hormones, ovary histomorphology and ZP3 antibody immunohistochemistry. The histomorphology and expressions of Ki-67, estrogen receptor α and estrogen receptor β in the uterus of the mice were detected.

RESULTSAutoimmune POP models were established successfully in 80% of the mice at 8 weeks after the immunization. Compared with those in the control group, the mice in the model group showed a smaller volume of the uterus, thinner endometrium and a reduced number of glands. The luminal epithelial cells, glandular epithelial cells and stromal cells in the uterus of the model mice all presented with a lower expression of Ki-67 than those in the control group, and Ki-67 translocation from the nuclei to the cytoplasm was found in the model group. The luminal epithelial cells, glandular epithelial cells and stromal cells showed positive ERα immunoreactivity in the model group but not in the control group. No obvious ERβ expression was found in the uterus in either of the groups.

CONCLUSIONpZP3 can induce autoimmune POP, cause suppressed proliferation of the endometrial epithelial cells and stromal cells, and reduce the cellular expression of ERα in the uterus of mice.

Animals ; Autoimmune Diseases ; metabolism ; Cell Nucleus ; Egg Proteins ; Endometrium ; Epithelial Cells ; Estrogen Receptor alpha ; metabolism ; Estrogen Receptor beta ; metabolism ; Female ; Immunohistochemistry ; Ki-67 Antigen ; metabolism ; Membrane Glycoproteins ; Mice ; Mice, Inbred BALB C ; Primary Ovarian Insufficiency ; metabolism ; Receptors, Cell Surface ; Stromal Cells ; Uterus ; metabolism ; Zona Pellucida Glycoproteins

Objective Compared with digital subtraction angiography(DSA),to evaluate the value of high-resolution contrast-enhanced three-dimensional magnetic resonance angiography(3D MRA) in defining hepatic arterial anatomy.Methods The data about abdominal high-resolution contrast-enhanced 3D MRA and DSA of 26 patients (24 patients with primary liver cancer,2 patients with metastatic liver tumor) was retrospectively analyzed.The display quality of different segmental hepatic artery was scored with 4 grades and the agreement between high-resolution 3D MRA and DSA was determined with the weighted Kappa statistic.The depiction of hepatic arterial anatomy/anomalies and vascular pathology on high- resolution 3D MRA was assessed and compared with DSA.Results With respect to display quality,there was good or fair correlation between high-resolution 3D MRA and DSA for the common hepatic artery (the mean score respectively was 3.96,3.96 and Kappa value 0.99),gastroduodenal artery (the mean score respectively 3.85,3.88 and Kappa value 0.85 ),right hepatic artery(the mean score respectively 3.92, 3.96 and Kappa value 0.65 ),left hepatic artery (the mean score respectively 3.77,3.92 and Kappa value 0.43 ),left gastric artery(the mean score respectively 3.73,3.85 and Kappa value 0.43 ),right anterior artery (the mean score respectively 3.35,3.70 and Kappa value 0.53),right posterior artery (the mean score respectively 3.31,3.73 and Kappa value 0.46)and 1V segment artery (the mean score respectively 2.92,3.46 and Kappa value 0.51 );Poor correlation was found for the Ⅱsegment artery (the mean score respectively 2.15,3.35 and Kappa value 0.18)and Ⅲsegment artery (the mean score respectively 2.19, 3.35 and Kappa value 0.21 ).Compared with DSA,18 normal hepatic arterial anatomy and 7 arterial anomalies were accurately demonstrated(accuracy ratio 96.1% (25/26)),the obliteration of gastroduodenal artery correctly depicted in 1 patient on high-resolution 3D MRA image.Conclusions High-resolution 3D MRA can provide accurate evaluation of hepatic artery and has the capacity of depicting hepatic segment artery.

Abstract: Objective　To construct SARS-CoV-2 receptor binding domain molecular probe for monoclonal memory B cell sorting and obtain RBD specific neutralizing antibodies from peripheral blood mononuclear cells (PBMCs) of COVID-19 convalescents by single-cell sorting. Methods　The SARS-CoV-2 RBD sequence was downloaded from GenBank, and the Avi tag and 6-histidine tags were added at the C-terminal. After codon optimization, it was chemically synthesized, cloned into the pDRVI1.0 vector, expressed after transfection of 293F cells, and biotinylated consequently. RBD-specific B cells were sorted out with this probe1 from the PBMCs of convalescents recovered from COVID-19. After B cells were lysed, the variable regions of heavy chain and light chain were amplified, cloned into the antibody expression vector, and transfected into 293F cells to express the antibody. Then the antibody was purified from the supernatant using protein A column and SARS-CoV-2 pseudovirus was used to test their neutralizing activity. Results　RBD-Avi probe was produced and successfully biotinylated sequentially with an efficiency of 30%-50%. Western blot analysis revealed that the biotinylated probe was recognized by the antibodies purified from COVID-19 convalescent plasma. Using this probe, 7 and 16 RBD-specific memory B cells were successfully isolated from the PBMCs of two convalescent individuals, accounting for 0.24% and 0.17% of the total cell population, respectively. After amplifying the variable regions of antibody heavy and light chains from the lysed B cells, 7 and 12 pairs of antibody heavy-light chains were obtained. A total of 16 antibodies were expressed in the convalescent individuals, and most of the purified antibodies showed neutralizing activity against the pseudovirus, with IC50 values of 6 antibodies below 1 μg/mL. The IC50 values of XJ-A9 and SCF-F1 against the wild-type pseudovirus were 0.07 μg/mL and 0.35 μg/mL, respectively. Conclusion　The SARS-CoV-2 RBD molecular probe constructed in this study has good antigenicity, and the isolated antibodies present neutralizing activity against wild-type SARS-CoV-2 pseudovirus.

Objective To explore the application of modified CT imaging scale in the assessment of prognosis of comatose patients with severe traumatic brain injury. Methods Retrospective analysis was performed on the clinical data of 90 comatose patients with severe traumatic brain injury; according to the prognosis,they were divided into conscious group (n=47) and coma group (n=43).The modified CT imaging scale was used to analyze the relationship between these CT imaging data and prognosis,and the relationship between these CT imaging data and severity of illness. Results The scores of CT imaging scale in comatose patients were significantly higher than those in conscious patients (P＜0.05). Patients with 3-5 scores in GCS enjoyed higher scores of CT imaging scale as compared with those with 6-8 scores (P＜0.05).The scores of modified CT imaging scale and the GCS scores in these 90 patients showed negative correlation (Spearman's correlation coefficient:r=-0.79,P=0.000). Conclusion The modified CT imaging scale is helpful in assessing the prognosis and guiding the clinical treatment of comatose patients with severe traumatic brain injury.

Objective To explore the rational approach for removing the meningomas located in the anterior cranial fossa. Methods Thirty-one cases with meningomas located in the anterior cranial fossa underwent the surgeries through combined pterional and subfrontal approach or unilateral or bilateral frontal approach from 2000 to 2005. Their total resection rates and complication incidences were compared retrospectively. Results Total resection was achieved in 15 of 16 cases using the combined pterional and subfrontal approach, and in 9 of 15 cases using the unilateral or bilateral frontal approach.Compared with the combined pterional and subfrontal approach, the incidence of complications was higher in the ones using the unilateral or bilateral frontal approach. Conclusion Combined pterional and subfrontal approach is a feasible and favorable approach for removing the meningioma located in the anterior cranial fossa.

OBJECTIVETo explore germline mutations of MLH1 in hereditary nonpolyposis colorectal cancer (HNPCC), and to investigate the pathobiology of novel detectable mutations of MLH1.

METHODRNA was extracted from the peripheral blood of 12 patients from 12 different families fulfilling the Amsterdam II Criteria of HNPCC. Germline mutations of MLH1 were determined by RT-PCR with gene specific primers, heat-resistance reverse transcriptase and long-template PCR polymerase, followed by cDNA sequencing analysis. PCR-Genescan analysis was used to further investigate microsatellite instability with a panel of 5 microsatellite markers (BAT26, BAT25, D5S346, D2S123 and Mfd15), along with immunohistochemistry staining to detect the expression of MLH1 protein in the tumor tissues.

RESULTSFour germline mutations were found in 4 patients, 2 of which were previously reported GTT-->GAT mutation at codon 384 of exon 12, and the other two were novel mutations: CGC-->TGC at codon 217 of exon 8 and CCG-->CTG at codon 581 of exon 16. Two tumors with the novel mutations had high frequency microsatellite instability showing more than 2 instable loci (RER + phenotype), and both tumors lost their MLH1 protein expression.

CONCLUSIONThe two novel germline mutations of MLH1 identified in this study, i.e. CGC-->TGC at codon 217 of exon 8 and CCG-->CTG at codon 581 of exon 16, are very likely to have pathological significance.

Adaptor Proteins, Signal Transducing ; Carrier Proteins ; genetics ; metabolism ; Codon ; Colorectal Neoplasms, Hereditary Nonpolyposis ; genetics ; metabolism ; DNA Mutational Analysis ; DNA, Neoplasm ; genetics ; Exons ; Female ; Germ-Line Mutation ; Humans ; Male ; Microsatellite Instability ; Middle Aged ; MutL Protein Homolog 1 ; Nuclear Proteins ; genetics ; metabolism ; Phylogeny