Objective To compare the body fluid count results detected by Sysmex XE-5000 automatic blood cell analyzer and manual method .Methods A total of 300 cases of body fluid specimens (including cerebrospinal fluid and fluid of serous cavity ) were analyzed .RBC ,WBC counting and classification were respectively detected by XE-5000 and manual method of improvement Neubauer counting plate .Results The fresh specimens without contain a large number of cell clusters ,which RBC counts(RBC-BF)(100-10 000)× 106/L ,and WBC counts(WBC-BF) (9-50)× 106/L ,showed there were a linear relationship between the in-strument method(r=0 .998 5 ,0 .986 3) .In the range ,there was no significant difference between XE-5000 and manual method(t=9 .96 ,P>0 .05) .Also in this range the results of instrument correlated with those of manual method(r= 0 .989 3 ,0 .971 7 , 0 .924 9) .For those specimens which contain a large number of cell clusters ,RBC-BF and WBC-BF were a badly linear relationship between the instrument method(r=0 .564 8 ,0 .456 1) .Conclusion Body fluid specimens which are fresh and do not contain a large number of cell clusters ,in the range of RBC-BF (100 -10 000) × 106/L ,WBC-BF (9 -50) × 106/L ,Sysmex XE-5000 automatic blood cell analyzer could ensure the results have good accuracy .

Objective To establish an ELISA method for detection of the concentration of TAP in human urine samples.Methods TAP-BSA was used as coating antigen. The TAP was a competitor to TAP-BSA. They reacted to the limited amount of monoclonal antibody against TAP.Results The optimal concentration of the coating antigen was 250ng/ml.The dilutions of monolclonal antibody against TAP and sheep anti-mouse IgG were 25?g/ml and 1∶4 000 respectively. The optimal range was from 0.69 to 1 000ng/ml.The assay provided a sensitivity of 0.69ng/ml.The coefficients of variation of intra-assay and inter-assay were 9.10% and 10.33% respectively. The average recovery rate of TAP was 97.70%.Conclusion The indirect competitive enzyme-linked immunosorbent assay is established.

Animals ; Atherosclerosis ; Humans ; Serine Proteinase Inhibitors ; Serpins

Objective To evaluate the performance of a DNA microarray method for detecting HBV antiviral drug-resistant mutations. Methods Two hundred and twenty four serum samples from patients with CHB were tested in parallel by DNA microarray and direct sequencing for the mutations within the HBV reverse transcriptase (rt) region, which included rtL180, rtA181, rtM204 and rtN236. Samples with discrepant results were retested by clonal sequencing. Results Complete concordance between DNA microarray and direct sequencing results was observed in 214 out of 224 samples (95. 5% ). The presence of mixed viral populations in the other 10 samples detected by DNA microarray but not by direct sequencing was confirmed by clonal analysis. The DNA microarray could detect minor viral populations which constituted 5.0%-15. 0% of the total viral load. Conclusion DNA microarray is highly consistent with direct sequencing in detecting HBV mutations conferring drug resistance and more sensitive in detecting mixed mutant and wild-type sequences than direct sequencing, which makes it a useful tool for early detection of drug resistance early.

Objective To study the clinical features of follicular occlusion triad, and whether it is a hereditary disease. Methods Based on the clinical examination of a case who developed squamous cell carcinoma secondary to follicular occlusion triad, the pedigree of the patient was surveyed and analyzed. Results There were a total of thirteen patients in this pedigree, the age of onset was about 20 years old. The clinical features and laboratory examination of the proband was consistent with follicular occlusion triad. Conclusions Hereditary factor is important in the pathogenesis in follicular occlusion triad,and the disease maybe an autosomal dominant inherited disease.

Objective To observe the intervention effect of electro-acupuncture(EA) on mammalian target of rapamycin(mTOR) expression in hypoxic-ischemic encephalopathy(HIE) newborn rats. Methods Seven-day-old SD rats were randomly assigned into 3 groups, namely sham-surgery group, HIE model group, and EA group. All of the groups were subdivided into 4 subgroups in terms of 4 time periods of 1, 3, 7 and 21 day(s). Hematoxylin-eosin staining was used for the observation of histological changes in the affected cerebral region. Expression levels of cerebral mTOR were detected with Western blotting method. Results The results of HE staining showed that EA group at experimental day 21 had clear organizational structure, ordered arrangement of neurons, relieved cellular swelling, well cell integrity with clear nucleolus and proliferative glial cells. Compared with the sham-surgery group, mTOR level of the model group was dramatically increased on the 1st, 3rd, 7th and 21st day(P<0.05), and the mTOR expression level of EA group was significantly higher than that in the model group at the above time points (P < 0.05). Conclusion EA can promote mTOR expression in brain tissue and can protect the brain of newborn rats from HIE.

AIM: To assess the impact of diabetes on corneal endothelial cells through the quantitative analysis of corneal endothelial cell morphology for patients with diabetics.
METHODS: The corneal thickness and endothelial cell morphology of 360 eyes of 299 cases were detected using full automatic corneal endothelial cell analyzer. The normal control group included 175 eyes of 148 cases, and there were 185 eyes of 151 cases for the patients with diabetes, 110 eyes of 92 cases for the non-proliferating phase group and 75 eyes of 59 cases for the proliferating phase group. The average density of central corneal endothelial cells, proportion of hexagonal cells, coefficient of variation and corneal thickness were compared among groups, and then the statistical analysis was conducted.
RESULTS: Compared with the cornea of the normal group, in the diabetes group, the coefficient of variation of corneal endothelial cells and central corneal thickness increased, while the average density of central corneal endothelial cells and proportion of hexagonal cells decreased, showing a significant difference (P<0. 05). Compared with the cornea of non-proliferating phase group, in the proliferating phase group, the density of central corneal endothelial cells decreased, the coefficient of variation of corneal endothelial cells increased, while the proportion of hexagonal cells decreased with a significant difference (P<0. 05), and the central corneal thickness increased, showing no significant difference(P>0. 05).
CONCLUSION: Compared with the cornea of normal control group, in the diabetes group, the corneal endothelial cells show abnormal morphology, which aggravates with the severity of lesions, especially for the significant changes in the coefficient of variation and the proportion of hexagonal cells. As a result, the corneal resistance to damage in patients with diabetes will decrease.

Objective To investigate the molecular mechanism of bone marrow mesenchymal stem cells (BM-MSCs) in repairing cisplatin-induced acute renal injury.Methods The rats were injected 6 mg/kg of cisplatin intraperitoneally,and bone marrow mesenchymal stem cells (BM-MSCs group) or PBS (PBS group) were injected respectively via tail vein after 24 hours.The rats without injecting cisplatin were selected as a normal control group.The repair effect of BM-MSCs on renal injury was observed by HE staining and immunohistochemistry.In addition,NRK-52E cells were cultured in vitro and treated with cisplatin for 6 hours.Then,NRK-52E cells were continued to culture for 48 hours or co-cultured with BM-MSCs for 48 hours,and NRK-52E cells untreated with cisplatin were used as a control.The expression levels of miR-92b and its target gene PTEN were detected by qRT-PCR,and the expression level of p-Akt by western blot.Results HE staining showed that the tubular protein casts in BM-MSCs group were significantly less than that in PBS group,and that the renal tubular structure was significantly improved in BM-MSCs group.Immunohistochemical staining indicated that the number of cells expressing proliferating cell nuclear antigen (PCNA) in BM-MSCs group (131.0 ± 14.4) was significantly higher than that in PBS group (42.2 ±6.1,t =11.28,P ＜0.01).qRT-PCR results showed that in the vivo experiment,compared with the expression level of miR-92b and PTEN in the normal control group (1.11 ± 0.78,1.01 ± 0.21),PBS group were (4.64 ± 1.06) and (0.61 ± 0.2),respectively (all P ＜ 0.05);BM-MSCs group were (2.27 ± 0.81) and (1.1 ± 0.1),respectively (all P ＜ 0.05).In vitro experiment,compared with the expression level of miR-92b and PTEN in the negative control group (1.12 ± 0.77,1.02 ± 0.13),cisplatin group were (7.64 ± 0.72) and (0.58 ± 0.2),respectively (all P ＜ 0.05),cell group were (4.38 ± 0.50) and (1.15 ± 0.23),respectively (all P ＜ 0.05).Western blot results showed that compared with the expression level of p-Akt in cisplatin group (0.96 ± 0.18),p-Akt expression in cell group was (2.11 ± 0.11,P ＜ 0.01).Conclusion BM-MSCs may repair the cisplatin-induced acute renal injury via down-regulating the expression level of miR-92b.

Objective To evaluate the etiologies of children with gastrointestinal bleeding(GIB), and the relationship between the pathological findings and clinical.Methods Gastroscopy or proctoscope was performed in 153 children with GIB. Pathological studies and tests as for helicobactor pylori (Hp) were undertaken.Results Among 153 children,140 cases(91.5 %) had definite diagnosis,including 74 children with upper gastrointestinal bleeding(UGIB) and 66 cases with lower gastrointestinal bleeding(LGIB). Gastric pathologic study was conducted in 56 cases. All patients had chronic superficial gastritis(CSFG). Hp test was positive in 33 patients. There was significant difference in the positive rate of Hp test between patients with gastritis or duodenitis and those with ulcers (P

Objective To investigate the characteristics of children′s recurrent abdominal pain(RAP) with gastroscopy and analyze the etiology.Methods Three hundred and thirteen children were investigated with gastroscopy.Gastric antrum mucosa was taken for histo-pathology and the determination of helicobacter pylori(Hp) antibody.Results Ninty-nine point six eight percent of the cases had lesions under the gastroscope.The former 4 cases had chronic superficial gastritis(CSG),CGS and bile reflux,CGS and duodenitis,CGS and bulb ulcer,and Hp infection rate was 31.36%,25%,38.64%,60.61%.Hp infection rates of active gastritis and inactive gastritis were 92% and 23.19%,which showed significant differences in 2 groups(P