OBJECTIVETo study the inhibitory effect of small interference RNA (siRNA) targeting cyclin A2 gene on the growth of osteosarcoma MG-63 and human normal skin fibroblast HSF cells and to explore whether cyclin A2 siRNAs could become a useful tool in the treatment of osteosarcoma.

METHODSThree pairs of siRNAs targeting cyclin A2 mRNA and a pair of nonsense siRNA were designed according to the current criteria. SiRNAs were chemically synthesized and purified. The siRNAs were transfected into MG-63 cells and HSF cells via oligofectamine. The cells transfected with nonsense siRNA served as negative control group and those only treated with PBS as blank control group. Quantitative fluorescence RT-PCR, Western-blot, MTT assay, reverse transcriptase (RT)-PCR, flow cytometry and clone forming test were employed to evaluate the efficacy of RNA interference. At the same time, the mRNA expression of PCNA and cyclin B1 in siRNA-treated MG-63 cells were examined.

RESULTSAlthough all three siRNAs could reduce the cyclin A2 expression, siRNA, appeared to be the most effective. After 48 h treatment with siRNA1, cyclin A2 mRNA and protein expression in MG-63 cells was significantly reduced by nearly 80% as compared with that of the blank control group, whereas the negative and blank control groups had similar expression levels. MG-63 cells treated with siRNA1 were arrested at G0/G1 phase by 80.1% and the proliferation of these tumor cells was suppressed 48 h after transfection. Furthermore, MG-63 cells showed a decreased colony forming ability after siRNA1 treatment. In addition, the cyclin A2-depleted MG-63 cells showed decreased levels of PCNA and cyclin B1. In contrast, although cyclin A2 expression in HSF reduced by nearly 60% after treatment by siRNA1 for 48h, these cells exhibited only a slight change in cell cycling, and neither clear inhibition of proliferation nor impaired colony forming ability was observed.

CONCLUSIONCyclin A2 is critical for proliferation of MG-63 cells. Cyclin A2-siRNAs can induce obvious inhibition of cyclin A2 mRNA and protein expression in MG-63 and HSF cells, which consequently down-regulate the proliferation of MG-63 cells. There is little effect on the proliferation of siRNA-treated HSF cells. Those results indicate that siRNAs against cyclin A2 may become a potential antiproliferative tool in future antitumor therapy.

Bone Neoplasms ; metabolism ; pathology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Cyclin A2 ; genetics ; metabolism ; Cyclin B1 ; metabolism ; Fibroblasts ; cytology ; metabolism ; Gene Knockdown Techniques ; Humans ; Osteosarcoma ; metabolism ; pathology ; Proliferating Cell Nuclear Antigen ; metabolism ; RNA Interference ; RNA, Messenger ; RNA, Small Interfering ; Skin ; cytology ; Transfection

This study aimed to quantitatively assess the effectiveness of the Wuhan lockdown measure on controlling the spread of coronavirus diesase 2019 (COVID-19). : Firstly，estimate the daily new infection rate in Wuhan before January 23，2020 when the city went into lockdown by consulting the data of Wuhan population mobility and the number of cases imported from Wuhan in 217 cities of Mainland China. Then estimate what the daily new infection rate would have been in Wuhan from January 24 to January 30th if the lockdown measure had been delayed for 7 days，assuming that the daily new infection in Wuhan after January 23 increased in a high，moderate and low trend respectively (using exponential, linear and logarithm growth models). Based on that，calculate the number of infection cases imported from Wuhan during this period. Finally，predict the possible impact of 7-day delayed lockdown in Wuhan on the epidemic situation in China using the susceptible-exposed-infectious-removed (SEIR) model. : The daily new infection rate in Wuhan was estimated to be 0.021%，0.026%，0.029%，0.033% and 0.070% respectively from January 19 to January 23. And there were at least 20 066 infection cases in Wuhan by January 23，2020. If Wuhan lockdown measure had been delayed for 7 days，the daily new infection rate on January 30 would have been 0.335% in the exponential growth model，0.129% in the linear growth model，and 0.070% in the logarithm growth model. Correspondingly，there would have been 32 075，24 819 and 20 334 infection cases travelling from Wuhan to other areas of Mainland China，and the number of cumulative confirmed cases as of March 19 in Mainland China would have been 3.3-3.9 times of the officially reported number. Conclusions: Timely taking city-level lockdown measure in Wuhan in the early stage of COVID-19 outbreak is essential in containing the spread of the disease in China.
COVID-19 ; China/epidemiology* ; Cities ; Communicable Disease Control ; Humans ; SARS-CoV-2

BACKGROUNDDicycloplatin is a relatively safe third generation platinum-complex anti-cancer drug. The present study focused on the effects of dicycloplatin on in vitro proliferation and apoptosis of human aortic smooth muscle cells (HASMC) and human aortic endothelial cells (HAEC).

METHODSProliferation of HASMC and HAEC, DNA content, and cellular levels of proliferation- and apoptosis-related proteins were assessed using the (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) (MTS) assay, flow cytometry and Western blotting assays, respectively.

RESULTSDicycloplatin at 10 ng/ml significantly inhibited HASMC proliferation, however, 10 µg/ml were required to significantly inhibit HAEC proliferation. Cell cycle analysis showed that dicycloplatin was a non-specific inhibitor of the cell cycle. Although dicycloplatin significantly decreased proliferating cell nuclear antigen (PCNA) expression in HASMC at all concentrations tested, it did not significantly affect PCNA expression in HAEC; Bax and p53 protein expression was upregulated in dicycloplatin groups.

CONCLUSIONSDicycloplatin at nanogram concentrations significantly inhibits HASMC proliferation, although the effect is relatively weaker than that of sirolimus. In contrast, the effect of dicycloplatin on inhibition of HAEC proliferation is much less pronounced than that on HASMC. The latter characteristics point to the potential for use of dicycloplatin in drug-eluting stents.

Aorta ; cytology ; Blotting, Western ; Cell Proliferation ; drug effects ; Drug Combinations ; Drug-Eluting Stents ; Endothelial Cells ; cytology ; drug effects ; Flow Cytometry ; Glutamates ; pharmacology ; Humans ; Muscle, Smooth, Vascular ; cytology ; drug effects ; Organoplatinum Compounds ; pharmacology ; Sirolimus ; pharmacology

To evaluate and summarize the evidence of diet and physical activity management in patients with metabolic syndrome (MS). BMJ Best Practice, UpToDate, Joanna Briggs Institute (JBI) database, Agency for Healthcare Research and Quality (AHRQ) network, National Institute for Health and Clinical Excellence (NICE) network, Scottish Intercollegiate Guidelines Network (SIGN), Guidelines International Network (GIN), Medlive, Registered Nurses' Association of Ontario (RNAO) network, American Diabetes Association (ADA) network, New Zealand Guideline Group (NZGG) network, Canadian medical association clinical practice guidelines network, PubMed, EmBase, Web of Science, CINAHL, Cochrane Library, CNKI, China Science and Technology Journal Database, Wanfang Knowledge Data Service Platform and Chinese biomedical database were searched systematically to obtain guidelines, evidence summary, expert consensus, best practice information book, clinical decision-making, recommended practice, and systematic review on diet and physical activity management in patients with MS. The retrieval period is from the establishment of database to November 2021. Two researchers with evidence-based medicine background evaluated the quality and evidence level of the included literature. A total of 36 articles met the criteria, including 3 guidelines, 5 expert consensus, 1 clinical decision and 27 systematic reviews. We summarized 49 pieces of evidence related to diet and physical activity in patients with MS, involving 15 aspects, namely diet goals, diet patterns, diet time, carbohydrate intake, fat intake, fiber intake, salt intake, fruits, vegetables and grains intake, coffee intake, effects of diet, principle of physical activity, intensity, form, time of physical activity, effects of physical activity, physical activity prescription of patients with MS and cardiovascular disease, and the joint effects of diet and physical activity. Diet and physical activity management can effectively improve the health outcomes of patients with MS. Health professionals should choose and apply the best evidence with consideration of the clinical situation and patient preference.
Canada ; Consensus ; Diet ; Exercise ; Humans ; Metabolic Syndrome/therapy*

OBJECTIVES: To develop a Chinese version of the Long-Term Conditions Questionnaire (LTCQ) and to test its reliability and validity in Chinese patients with chronic diseases. METHODS: With the consent of the original authors, a Chinese version of LTCQ was developed according to the cultural adjustment guidelines. A questionnaire survey was conducted on 319 patients with chronic diseases in Sir Run Run Shaw Hospital, Wuyi County First People's Hospital and Hangzhou Gongchen Bridge Street Health Service Center. The questionnaire was evaluated by item analysis (including frequency analysis, total question correlation method and critical ratio method), reliability analysis (Cronbach's alpha coefficient) and validity analysis [including content validity (expert scoring method) and structural validity (exploratory factor analysis)]. RESULTS The Chinese version of the LTCQ included 20 entries, with a Cronbach's alpha coefficient of 0.926, a retest reliability of 0.829, a split-half reliability of 0.878, an entry content validity index of 1, and a content validity index at the questionnaire level of 1. Four common factors were extracted by exploratory factor analysis, namely physical state and daily life, psychological state, support and coping, and safe environment, with a cumulative variance contribution rate of 67.244%. Discussion: The Chinese version of the LTCQ developed in this study has good reliability and validity and it may be used to assess the long-term conditions of patients with chronic diseases in China.
Humans ; Asian People ; China ; Chronic Disease ; Quality of Life ; Reproducibility of Results ; Surveys and Questionnaires

OBJECTIVETo study the pharmacokinetic of 10-Hydroxycamptothecin HCPT by intraperitoneal administration.

METHODSix dogs were given HCPT by intraperitoneal perfusion: HCPT 2 mg x kg(-1), dissolved in 300 mL NS. During the course of experiments, peritoneal fluids and blood were sampled using a standardized protocol. The concentration of HCPT in all samples was determined by high performance liquid chromatography (HPLC).

RESULTThe area under the curve (AUC) of peritoneal fluids was significantly highly as compared to the AUC of femoral vein and portal vein (P < 0.001).

CONCLUSIONThese experiments demonstrate that intraperitoneal chemotherapy has significant effect on the pharmacokinetics of IP route of HCPT administration. This method should be a more reasonable chemotherapy for the prevention of recurrence and liver metastasis of gastric cancer, colon cancer and ovary cancer after radical resection.

Animals ; Antineoplastic Agents, Phytogenic ; administration & dosage ; pharmacokinetics ; Area Under Curve ; Ascitic Fluid ; metabolism ; Camptothecin ; administration & dosage ; analogs & derivatives ; pharmacokinetics ; Dogs ; Infusions, Parenteral ; Male

OBJECTIVETo compare the performance of full-field digital mammography (FFDM) and digital breast tomosynthesis (DBT) in the assessment of the lesions in dense breast, and to estimate the difference in diagnosis of breast disease by FFDM images alone and FFDM plus DBT images.

METHODSAccording to the breast imaging reporting and data system (BIRADS), 134 patients were selected. The morphology of the lesions shown on FFDM and DBT were evaluated and compared, and the maximum diameter of the lesions was measured. At first, doctors made the diagnosis of a patient by reading FFDM only. Then they made another diagnosis by combining with DBT images of the same patient. The two diagnoses were compared and analyzed according to the pathology results.

RESULTSOne hundred and thirty-four patients were included in this study, and all of them were confirmed by histology (65 benign cases, 69 malignant cases). DBT could show more details about the morphology of the lesions, including the border of the masses, spiculation and vessels. The numbers of those signs detected by DBT were 46, 30 and 3, respectively, while only 33 case with circumscribed masses and 14 cases with spiculation were detected by FFDM. Only the difference of spiculation in heterogeneously dense breast detected by DBT and FFDM was statistically significant (P < 0.05). Of the cases with calcifications, DBT images (reconstructed as a 1-mm-thick slice) showed calcifications superior to FFDM in 2 cases, equal to FFDM in 23 cases, and inferior to FFDM in 11 cases. The difference was statistically significant (P < 0.05). But when thickness was changed into 1 cm, the visibility of calcifications in those cases was equal between FFDM and DBT. The maximum diameter of lesions was 2.46 ± 1.64 cm in DBT image, and 2.58 ± 1.62 cm in FFDM image, with a significant difference (P < 0.05). Comparing with reading FFDM images only, the accuracy of FFDM combining with DBT was increased from 88.8% to 91.8%. For FFDM, the AUC of ROC was 0.887, while for DBT it was increased to 0.912, with a non-significant difference (P > 0.05).

CONCLUSIONSDBT is superior to FFDM in the morphological characterization and small calcification in the lesions in dense breast. Combining FFDM and DBT improves the accuracy of diagnosis, but the difference is not statistically significant.

Adolescent ; Adult ; Breast Diseases ; diagnostic imaging ; Breast Neoplasms ; diagnostic imaging ; Calcinosis ; diagnostic imaging ; Carcinoma in Situ ; diagnostic imaging ; Carcinoma, Ductal, Breast ; diagnostic imaging ; Female ; Fibroadenoma ; diagnostic imaging ; Humans ; Image Processing, Computer-Assisted ; Imaging, Three-Dimensional ; Mammography ; methods ; Middle Aged ; Radiographic Image Enhancement ; Tomography, X-Ray Computed ; Young Adult

OBJECTIVEThis prospective study was to observe the correlation between the mesial papilla's height of single implant-supported maxillary central incisor and the distance from the base of the contact point to the alveolar bone crest.

METHODS56 patients involved in single implant-supported maxillary central incisor were included in this study. The distances from the base of the contact point to the alveolar bone crest in the digital periapical film of maxillary central incisor were measured using the software Planmeca Dimaxis Version 3.3.2. The time of measurements were as follows: The pre-surgical and post-surgical periods, before and after the crown installation, the follow-up examination of more than 0.5 year. To analyze the factor of influencing distance, and the correlation between the distance and the height of gingival papilla during the whole restored period. Correlation analysis between the distance and the height of gingival papilla during the whole restoration was done by the statistical software SPSS 12.0.

RESULTSThe results demonstrated that the ratio of esthetic papilla can achieve 54.5% at the crown installation and 95.5% at the follow-up examination when the distance was between 3 mm and 5 mm. When the distance was between 5 mm and 6 mm, they dropped to 30.0% and 75.0%, respectively. However, when the distance increased to above 7 mm, the papilla could hardly be in an esthetic outcome. There was a significant change of the distance was found during the periods from the post-surgery to pre-restoration, and the scope of the changes was between -0.13 mm and 0.46 mm. A negative correlation was found between the distance and the index of papilla. The correlation coefficient r was -0.715 (P < 0.01).

CONCLUSIONIt is proposed that the pre-surgery distance of maxillary central incisor from the base of the contact point to the alveolar bone crest can be used as one of the important reference indexes to assess and predict the height conditions of gingival papilla.

Alveolar Process ; Anodontia ; Crowns ; Esthetics, Dental ; Gingiva ; Humans ; Incisor ; abnormalities ; Maxilla ; Prospective Studies

BACKGROUNDRecent studies have reported germline mutations in the perforin gene (PRF1) in some types of hemophagocytic lymphohistiocytosis (HLH). However, the prevalence of PRF1 mutations in HLH in Chinese pediatric patients has not been extensively studied. The aim of this study was to investigate the prevalence of mutations and sequence variations in the PRF1 gene in Chinese pediatric patients with HLH.

METHODSPolymerase chain reaction (PCR) was performed with five pairs of primers for the coding exons and the flanking intron sequences of PRF1. Sequencing of PCR products was subsequently applied in 30 pediatric patients with HLH and in 50 controls.

RESULTSThree heterozygous mutations in a coding region were found, which resulted in amino acid changes (C102F, S108N and T450M) in three patients. These mutations were not detected in control subjects. One patient had compound heterozygous mutations (S108N and T450M) in PRF1 as the background defect, and documented familial HLH type 2 (FHL2). One synonymous sequence variant (Q540Q) was observed in one patient but not in the controls. Two SNPs (A274A, H300H) in the coding region were detected in HLH patients and controls, but without differences in the heterozygosity rate between the two groups (P > 0.05 for all comparisons).

CONCLUSIONSWe have identified three patients with three heterozygous missense mutations in PRF1; two of those three mutations (C102F and S108N) have so far been found only from Chinese patients. These findings are useful in evaluating the prevalence of PRF1 mutations in Chinese pediatric patients with HLH, and to correlate their genotype with phenotype. Some patients without familial history probably have primary HLH, which should be suspected even beyond the usual age range.

Adolescent ; Amino Acid Sequence ; Child ; Child, Preschool ; Epstein-Barr Virus Infections ; genetics ; Female ; Humans ; Infant ; Lymphohistiocytosis, Hemophagocytic ; genetics ; Male ; Molecular Sequence Data ; Mutation ; Perforin ; Polymerase Chain Reaction ; Pore Forming Cytotoxic Proteins ; genetics

The aim of the study is to evaluate the effects of the single and mixed decoction of Thallus laminariae (kelp) and Glycyrrhiza glabra (licorice) on the metabolism and their difference. The mixed decoction of kelp and licorice and the single decoction were made and intragastrically administered to the SD rats. The effect on system metabolism, the toxicity of liver and kidney were assessed by GC-MS profiling of the endogenous molecules in serum, routine biochemical assays and histographic inspection of tissues from SD rats, separately. The mixed decoction of kelp and licorice induced more obvious pathological abnormalities in SD rats than a single decoction of kelp, while the extracts of licorice did not show any pathological change. Neither the mixed, nor the single decoction showed abnormal histopathology. After intragastric administration of extracts for 5 days, the mixed decoction induced a decrease of ALT (no significant change in the groups of single decoction) and an increase of BUN (so did the single decoction of kelp). Metabolomic profile of the molecules in serum revealed that the metabolic patterns were all obviously affected for the three groups, i.e., the mixed and single decoction of kelp and licorice. The rats given with the single decoction of kelp showed a similar pattern to that of the mixed decoction, indicating that the kelp primarily contributed the perturbation of metabolism for the mixed decoction. All three groups induced a decrease of branched chain amino acids, TCA cycle intermediates and glycolysis intermediates (e.g., pyruvic acid and lactic acid) and an increase of 3-hydroxybutyric acid. Kelp decoction showed stronger potential in reducing TCA cycle intermediates and glycolysis intermediates than the other two groups, while the levels of branched chain amino acids were the lowest after licorice extracts were given. These results suggested that the effect of the mixed decoction on metabolism was closely associated with both kelp and licorice. The continuous administration of single decoction of kelp and the mixed decoction of licorice and kelp resulted in pathological abnormalities in kidney of SD rats. The mixed decoction of kelp and licorice distinctly perturbed sera molecules and hence system metabolism, which showed associated with those of kelp and licorice. Although the metabolic effect was associated with both kelp and licorice, the results suggested kelp contributed to it primarily.
Animals ; Glycyrrhiza ; chemistry ; Kelp ; chemistry ; Kidney ; drug effects ; Liver ; drug effects ; Metabolomics ; Plant Preparations ; pharmacology ; Rats ; Rats, Sprague-Dawley