Objective To evaluate and select the dilution for high blood and urinary amylase among wet chemical & dry chemical de‐tection system ,hope to screen the optimum dilution for different measurement system and specimen type .Methods Pure water (H2 O) ,nor‐mal saline (NS) ,7% bovine serum albumin(7% BSA) ,low amylase specimen of serum and urine selected as candidate dilution .The high amylase specimen was diluted ,then was measured among wet chemical analyzer and dry chemical analyzer ,then the bias and correlation was evaluated according to the professional standard of National health and family planning commission of china .Results For high amylase ser‐um specimen ,the same dilution resulted in different bias between two detecting system .The bias of H2 O ,7% BSA and low amylase serum were less than the TEa (total error allowance) regulated by professional standard in wet chemical analyzer .Of them ,the low amylase serum was the best .In dry chemical analyzer ,the bias of H2 O ,NS and low amylase serum were no less than the professional standard .Of them ,NS was the best .For high amylase urine specimen ,the bias of H2 O and low amylase urine were less than the professional standard in wet chem‐ical analyzer ,and the low amylase urine was the best .In dry chemical analyzer ,only the bias of low amylase urine met the requirement of professional standard .The evaluation of linearity indicated that NS was unfit to the dilution for high amylase serum in wet chemical analyzer and for high amylase urine in dry chemical analyzer ,7% BSA was unsuitable to be the dilution for high amylase serum .Conclusion The effect of same dilution on different detecting system is different .The most suitable dilution is low amylase serum and urine for high amylase serum and urine specimen respectively in wet chemical analyzer ,and the suitable dilution is NS and low amylase urine respectively in dry chemical analyzer .

Objective To investigate the inhibitory effect of heparanase antisense oligodeoxynucleotide (AS-ODN) on the invasiveness of human liver cancer cell lines SMMC-7721,BEL-7402 in vitro. Methods We designed and synthesized AS-ODN that was complementary to the start codon region of heparanase mRNA,and the control,nonsense oligodeoxynucleotide (NS-ODN). The ODNs with the final concentration of 300 nmol/L was delivered into SMMC-7721,BEL-7402 cells by Oligofectamine TM Reagent. We evaluated heparanase gene expression using RT-PCR and detected heparanase protein expression using Western blot assay after transfection. Cell invasiveness was measured by Matrigel invasion assay.Results The result showed that heparanase gene expression,protein expression and invasiveness in AS-ODN group decreased significantly as compared with control groups( P

Objective To analyze the clinical characteristics and prognosis of disseminated cryptococcosis in children.Methods The data of disseminated cryptococcosis inpatients were reviewed retrospectively at Beijing Children's Hospital,Capital Medical University,from January 2002 to September 2014.The demographic data,clinical manifestations,laboratory findings,imaging,antifungal treatments and outcomes of all the patients were analyzed.Results Overall 25 children with disseminated cryptococcosis were enrolled including 17 boys(68.0％).The average age was 7 years old.Four cases (16.0％) had underlying conditions,among them 1 case had human immunodeficiency virus(HIV)-positive.The median time to diagnosis was 32 (23-47) days,44.0％ of the patients (11 cases) were misdiagnosed,and 8 cases misdiagnosed as tuberculosis.All patients had fever.Other common clinical manifestations included cough (16 cases),headache (10 cases),vomiting (10 cases),altered mental status (6 cases) and stomachache (6 cases).Respiratory system involvement was seen in all cases,central nervous system was involved in 18 cases,other organ involvement included lymph nodes,spleen,liver,kidney,skin,skeleton and costicartilage.Amphotericin B (AmB) + Fluconazole (Flu) ± 5-flucytosine (5-FC) was the most common therapy (15 cases),Flu ± 5-FC for 6 cases,AmB ± 5-FC for 2 cases,Voricanazole (VOR) for 1 case.Sixteen cases (66.7％) got recovery/improved on discharge,8 cases (33.3％) rejected to the advise and discharged with treatment failure,and the HIV-infected patient transferred to a special hospital.Fifteen patients (60.0％) were followed up,and 13 cases (87.0％) showed recovery,but 2 died in the long-term prognosis.Conclusions Disseminated cryptococcosis in children is more common in school-age boys.Most patients are without underlying conditions.Disseminated cryptococcosis can cause multiple organ damage.Patients with prolonged fever,cough,headache,with or without underlying diseases,can be suspected as disseminated cryptococcosis.Blood and cerebrospinal fluid culture for fungus,cryptococcal antigen should be performed for early diagnosis and treatment.

BACKGROUND:It is important to prepare a mal eable putty to repair irregular bone defects because the preformed bone substitute material cannot suit to bone defects.
OBJECTIVE:To prepare mal eable demineralized bone matrix/hyaluronic acid putty and to screen out the optimal composite ratio, as wel as to evaluate its cellcompatibility.
METHODS:Demineralized bone matrix was prepared from the cortical bone of healthy donors. The hyaluronic acid was dissolved at concentrations of 0.75%, 1.5%, 3%, 4.5%, and the viscosity of them was measured. Then 450 mg demineralized bone matrix was compounded with 1 mL hyaluronic acid solution to prepare the demineralized bone matrix/hyaluronic acid putty, and the optimal ratio of demineralized bone matrix/hyaluronic acid was screened by the disintegrated time of the putty at 37 ℃. L-929 mouse fibroblasts were cultured in the leaching liquor of demineralized bone matrix/hyaluronic acid, and the cytotoxicity was tested by cellcount kit-8 assay at days 1, 4 and 7.
RESULTS AND CONCLUSION:With the increase of hyaluronic acid concentration, the viscosity of hyaluronic acid solution was increased;the demineralized bone matrix/hyaluronic acid putty was strengthened and the disintegrated time was prolonged gradual y. When the ratio of demineralized bone matrix/hyaluronic acid was 15/1, demineralized bone matrix/hyaluronic acid was molded easily, and the disintegrated time was 8 hours in 7 ℃ PBS. The results of cytotoxicity showed that cellproliferation rates were 93.72%, 101.65%and 97.68%at days 1, 4 and 7 respectively, while the cytotoxicity was in grade 0 or 1. These findings indicate that demineralized bone matrix/hyaluronic acid putty at a compound ratio of 15:1 can be molded easily and difficult to disintegrate, with low cytotoxicity.

Objective To study the relationship between the abnormal blood fat components and the serum hypersensitivity C reaction protein concentration and the progression of diabetic retinopathy.Methods Fundus examination was carried out to inpatients in the Department of Endocrinology in the hospital from From July 2015to July 2017.These patients were divided into three groups according to results of fluorescein sodium contrast:The non retinopathy group (group A) , the non proliferative retinopathy group (group B) , the retinopathy proliferative phase group (group C) .The content of blood fat component and serum hypersensitivity C reaction protein in the experimental cases were measured, and the ratio of TC/HDL-C was calculated.The statistic software of SAS9.4was used for statistical processing.Results The concentration of the serum hypersensitive C reaction protein in patients with diabetic retinopathy was significantly increased and the ratio of TC/HDL-C increased significantly (P<0.05) .The hs-CRP concentration and TC/HDL-C ratio of the three study groups were compared with those of the healthy control group:the difference was statistically significant (P<0.05) .Among them, the hs-CRP concentration and TC/HDL-C ratio of group A were compared with group B and group C respectively and the difference were statistically significant (P<0.05) .There was no significant difference in hs-CRP concentration and TC/HDL-C ratio between group B and group C (P>0.05) .Linear correlation analysis was used to analyze the concentration of serum hypersensitive C reaction protein and the ratio of TC/HDL-C.It was found that there was a positive correlation between them, and there was a positive synergistic effect.Conclusion The measurement of serum hypersensitive C reactive protein has important clinical value for the discovery, evaluation and prognosis of diabetic retinopathy.The abnormalities of the blood fat component reflected by the TC/HDL ratio are also an important factor in assessing the progression of diabetic retinopathy.

Objective To investigate the mechanism that receptor activator of NF-κB ligand (RANKL) promotes arterial calcification.Methods Firstly,RANKL was added into the culture media,in which the monocyte precursor cells alone were cultured.Morphological observation and tartrate resistant acid phosphatase(TRAP)stain were used to assess whether RANKL could induce the monocyte precursor cells to differentiate into osteoclast-like cells.During arterial calcification,both in vivo and in vitro expressions of RANKL and osteoprotegerin (OPG,as RANKL inhibitor)were measured via real-time PCR.The extent of osteoclast-like cell differentiation was also assessed.Results It was found that RANKL could induce osteoclast-like cell differentiation.There were no both in vivo and in vitro expressions of osteoclast-like cells in the early stage of calcification.At that time,the ratio of RANKL to OPG was very low.In the late stage of calcification,a small amount of osteoclast-like cell expression coincided with a relatively high ratio of RANKL to OPG.According to the results,the ratio of RANKL to OPG was very low during most of the arterial calcification period.This made it possible for OPG to completely inhibit RANKL-induced osteoclast-like cell differentiation.The ratio of RANKL to OPG was (0.36 ± 0.08) (F =36) and (1.68 ± 0.08) (F =36) respectively in the early and late subgroup of calcification group in the animal model,but was zero in the control group(both P＜0.05).The ratio of RANKL to OPG was(0.42±0.09) (F=16)and(1.50 ± 0.10)(F=16)respectively in the early and late subgroup of calcification group in the cell model,but was zero in the control group(both P＜0.05).Conclusions Our result likely explains why RANKL has the ability to induce osteoclast-like cell differentiation,but acts as a promoter of calcification.

Objective To provide scientific evidence to identify and confirm severe acute respiratory syndrome (SARS) by laboratory detection.Methods Multiple clinical specimens were collected serially and systematically from the 4 suspected SARS patients, which occurred between Dec.2003 to Jan.2004 in Guangdong Province. The samples were tested by serologic and molecular methods.Results IgM or IgG antibodies against SARS-CoV were detectable after 6—8 days of the onset in four patients. The four-fold or greater rising in antibodies was clearly detected in three of the four patients, while the fourth patient’s seroconversion was from negative to positive. The results analysed by enzyme-linked immunosorbent assay( ELISA), immunoflourescence assay (IFA), and neutralization test were highly correlated. SARS-CoV RNA was just detected in 3 throat swab specimens from case 1 by real-time PCR. M, N and S genes were amplified by reverse transcriptase polymerase chain reaction (RT-PCR) from the positive samples. Sequencing results showed that they were SARS-CoV gene segments, and most closely matched SARS-CoV gene sequences were isolated from civet cats in Guangdong Province. Nevertheless, SARS-CoV was not isolated from any samples of the 4 patients.Conclusion Based on these results, the 4 reported cases were laboratorily confirmed as SARS cases.

Enterotoxigenic Escherichia coli（ETEC）infection can induce watery diarrhea，leading to dehydration，electrolyte disturbance，and even death in severe cases. Recombinant B subunit/inactivated whole-cell cholera（rBS/WC）vaccine is effective in preventing ETEC infectious diarrhea. On the basis of the latest evidence on etiology and epidemiology of ETEC，as well as the effectiveness，safety，and health economics of rBS/WC vaccine，National Clinical Research Center for Child Health（The Children’s Hospital，Zhejiang University School of Medicine）and Zhejiang Provincial Center for Disease Control and Prevention invited experts to develop expert consensus on rBS/WC vaccine in prevention of ETEC infectious diarrhea. It aims to provide the clinicians and vaccination professionals with guidelines on using rBS/WC vaccine to reduce the incidence of ETEC infectious diarrhea.