AIM: To explore the feasibility of culturing human umbilical vein endothelial cells ( HUVEC ) on acellular corneal stroma and performing the posterior lamellar endothelial keratoplasty ( PLEK ) treating corneal endothelial decompensation.
METHODS: Thirty New-Zealand rabbits were divided into three groups randomly, 10 rabbits for experimental group, 10 for stroma group and 10 for control group. Corneal endothelial cells were removed to establish animal model of corneal endothelial failure. PLEK was performed on the rabbits of experimental group and stroma group, and nothing was transplantated onto the rabbits of control group with the deep layer excised only. Postoperative observation was taken for 3mo. The degree of corneal edema and central corneal thickness were recorded for statistical analysis.
RESULTS:Corneas in experimental group were relieved in edema obviously compared with that in stroma group and the control group, and showed increased transparency 7d after the operation. The average density of endothelial cells was 2 026. 4±129. 3cells/mm2 , and average central corneal thickness was 505. 2±25. 4μm in experimental group, while 1 535. 6±114. 5μm in stroma group and 1 493. 5±70. 2μm in control group 3mo after operation.
CONCLUSION:We achieved preliminary success in our study that culturing HUVEC on acellular corneal stroma and performing PLEK for corneal endothelial decompensation. HUVEC transplanted could survive in vivo, and have normal biological function of keeping cornea transparent. This study provides a new idea and a new way clinically for the treatment of corneal endothelial diseases.

Objective To investigate the clinical significance of serum pepsinogen (PG) in gastric cancer screening. Methods The clinical data of 930 patients underwent colonoscopy were retrospectively analyzed. Among them, non chronic atrophic gastritis was in 550 cases (chronic atrophic gastritis group), chronic atrophic gastritis in 300 cases (chronic atrophic gastritis group), gastric cancer in 80 cases (gastric cancer group). The patients in chronic atrophic gastritis group were divided into mild chronic atrophic gastritis subgroup (100 cases), moderate chronic atrophic gastritis subgroup (120 cases) and severe chronic atrophic gastritis subgroup (80 cases) according to the severity of the atrophy. The levels of serum PGⅠand PGⅡwere detected by enzyme linked immunosorbent assay (ELISA) method, and the ratio of PGⅠand PGⅡ(PGR) was calculated. Results There was no statistical difference in PGⅡ among the 3 groups (F = 1.226, P>0.05). The PG Ⅰand PGR in gastric cancer group were significantly lower than those in chronic atrophic gastritis and non chronic atrophic gastritis:(70.41 ± 39.42)μg/L vs. (83.10 ± 30.08) and (165.5 ± 41.40)μg/L, 3.76 ± 2.03 vs. 5.08 ± 1.82 and 6.84 ± 1.88, those in chronic atrophic gastritis were significantly lower than those in non chronic atrophic gastritis group, there were statistical differences (P<0.05). The PG Ⅰand PGR in mild and moderate chronic atrophic gastritis subgroup were significantly higher than those in severe chronic atrophic gastritis subgroup and gastric cancer group:(95.50 ± 30.80) and (82.10 ± 31.42)μg/L vs. (70.12 ± 20.12) and (70.41 ± 39.42) μg/L, 5.84 ± 2.88 and 5.08 ± 1.89 vs. 3.90 ± 2.78 and 3.76 ± 2.03, there were statistical differences (P<0.05), but there was no statistical difference between severe chronic atrophic gastritis subgroup and gastric cancer group (P>0.05), and there was no statistical difference between mild chronic atrophic gastritis subgroup and moderate chronic atrophic gastritis subgroup (P>0.05). The receiver operating characteristic (ROC) curve was used, the optimal critical value of PG Ⅰ was 74.8μg/L, the area under curve (AUC) was 0.842, the sensitivity was 90%, specificity was 75%;the optimal critical value of PGR was 4.46, AUC was 0.837, the sensitivity was 75%, specificity was 82%;the AUC of combined detection of PG Ⅰ and PGR was 0.906, the sensitivity was 88%, specificity was 85%. Conclusions Detection of PG Ⅰ combined with PGR can be used as gastric cancer screening, the recommended level of PGⅠ≤74.80μg/L and PGR≤4.46.

Objective To investigate the diagnostic value of serum homocysteine in the diagnosis of colon cancer.Methods The performance rate method was used to detect the level of serum homocysteine(Hcy) in colon cancer group(50 cases) who were treated in Beijing Tiantan Hospital Affiliated to Capital Medical University from March 2011 to June 2016 and control group(50 cases).The expression of independent samples t test was used to analysis of the difference of the Hcy levels between the two groups.The ROC curve was used to evaluate the value of Hcy in diagnosis of colon cancer.Results The serum Hcy level in colon cancer group was (18.6±8.9) μmol/L,in healthy control group was (10.7±4.3) μmol/L,colon cancer group serum Hcy levels were significantly higher than those of healthy control group,there was significant difference(t=5.627,P<0.01).AUC of ROC curve was 0.775,cut-off value of 18.5 μmol/L,sensitivity was 0.50,specificity was 0.94,95%CI was 0.682-0.868(P<0.01).Conclusion Serum Hcy can be used as a reference index of the diagnosis of colon cancer.

Biomarkers, Tumor ; metabolism ; Carcinoma, Papillary ; metabolism ; Cathepsin B ; metabolism ; Cell Cycle Proteins ; metabolism ; HSP27 Heat-Shock Proteins ; metabolism ; Humans ; Proteomics ; Repressor Proteins ; metabolism ; S100 Calcium Binding Protein A6 ; S100 Proteins ; metabolism ; Serpins ; metabolism ; Thyroid Neoplasms ; metabolism

The RAS (rat sarcoma) gene is one of the important oncogenes, and its mutation is present in about 30% human tumors. KRAS (kirsten rat sarcoma viral oncogene) is one of the three RAS subtypes, and KRAS mutations are more common than the mutations in other two RAS subtypes. In recent years, with the continuous research, new ideas have been provided for the treatment of cancers via targeting-KRAS. Efforts have been made to develop various KRAS inhibitors. Here, based on the mechanism of action, we classified KRAS inhibitors into two categories: inhibitors that directly target KRAS and inhibitors that indirectly act on KRAS. The representative KRAS inhibitors were summarized and introduced in this paper.

Carcinoma in Situ ; epidemiology ; etiology ; pathology ; Carcinoma, Squamous Cell ; pathology ; surgery ; Disease Progression ; Female ; Humans ; Lichen Sclerosus et Atrophicus ; epidemiology ; etiology ; pathology ; Prevalence ; Vulvar Neoplasms ; epidemiology ; etiology ; pathology

Objective To explore the clinical significance of adenosine deaminase ( ADA) in pleural ef-fusion for diagnosis of tuberculosis pleuritis in children. Methods The level of ADA in pleural effusion was ret-rospectively analyzed in 28 cases with purulent pleuritis,thirty-four cases with mycoplasma pneumoniae pleuri-tis,forty-five cases with tuberculosis pleuritis from July 2011 to January 2014 in Beijing Children′s Hospital Af-filiated to Capital Medical University. Results The level of ADA in three groups was expressed by median (range interquartile). ADA in the purulent pleuritis group [126. 35 (76. 80,178. 13)U/L]was higher than the group of mycoplasma pneumoniae pleuritis [ 55. 55 ( 42. 80, 79. 03 ) U/L ] and tuberculosis pleuritis [ 26. 50 (22. 05,50. 95)U/L]. The difference was statistically significant (P< 0. 01). The cut-off value of pleural effu-sion ADA for diagnosis of tuberculosis pleuritis is not available by application of ROC curve. Conclusion Higher ADA value is not only the characteristic of tuberculosis pleuritis,but also purulent pleuritis and mycoplas-ma pneumoniae pleuritis. ADA has no clinical value in diagnosis of tuberculosis pleuritis in children.

BACKGROUND:There are a variety of treatments for femoral head necrosis,but their efficacy is not confirmed and unified.How to improve the differentiation ability of osteoblasts in the femoral head and improve the biomechanical support after the repair of the femoral head is an urgent problem to be solved.OBJECTIVE:To explore the clinical outcome of stem cells combined with vascularized iliac bone flap and tantalum rod implantation for the treatment of osteonecrosis of the femoral head (ONFH).METHODS:Totally 28 cases (36 hips) of non-traumatic ONFH admitted at the Zhongshan Hospital of Dalian University from January 2010 to January 2011 were enrolled.Bone marrow samples were extracted from each patient to isolate bone marrow stromal stem cells which were cultured in vitro for 2 weeks.Tantalum rod implantation with vascularized iliac bone graft was conducted to restore the femoral head shape,and then,prepared stem cell suspension were injected into the iliac bone flap and into the subchondral space of the femoral head.RESULTS AND CONCLUSION:All the 28 cases (36 hips) were followed up for 6-20 months (average 12 months),and their Harris hip scores and visual analogue scale scores at postoperative 6 and 12 months were significantly higher than the baseline (P ＜ 0.05).The Harris hip score at postoperative 12 months was significantly higher than that at postoperative 6 months (P ＜ 0.05),but there was no significant difference in the visual analogue scale scores at 6 and 12 months postoperatively (P ＞ 0.05).At the end of 12-month follow-up,clinical outcomes were excellent in 13 hips,good in 15 hips,fair in 4 hips,and poor in 4 hips,with an excellent and good rate of 90％.These findings indicate that autologous bone marrow stromal stem cell transplantation with vascularized iliac bone flap and tantalum rob implantation is an effective method with high clinical success rate for the treatment of ONFH.

BACKGROUND:Seed cells and scaffold are two key factors for cartilage defects after osteonecrosis of femoral head using tissue-engineered method. OBJECTIVE:To explore the feasibility of Bio-gide col agen membrane combined with dog bone marrow mesenchymal stem cells into chondrocytes. METHODS:Bone marrow mesenchymal stem cells were isolated from beagle dogs by whole bone marrow blood centrifugation method and adherence screening method in vitro and cultured. Morphological changes in cells were observed, and identification was done using cellsurface antigens. Bone marrow mesenchymal stem cells of passage 3 were induced by chondrocyte induction medium to differentiate into chondrocytes (experimental group). cells cultured in normal medium were considered as control group. 3-(4, 5-Dimethylthiazol-2-yl) 2, 5-diphenyl tetrazolium bromide assay was used to measure growth curve of chondrocytes. cells underwent typeⅡcol agen immunohistochemistry and toluidine blue staining. The coculture of bone marrow mesenchymal stem cells at passage 3 and Bio-gide col agen membrane were observed under an inverted phase contrast microscope and scanning electron microscope. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cells with high purity and high viability were obtained by whole bone marrow blood centrifugation method and adherence screening method. cells grew wel and had strong amplified ability, and successful y differentiated into chondrocytes. Numerous bone marrow mesenchymal stem cells adhered on the Bio-gide col agen membrane, showing a tendency of multi-layer growth. cells and Bio-gide col agen membrane seem to blend into an integrant part. Cel processes appeared and connected each other and gradual y wrapped the Bio-gide col agen membrane, with the presence of obvious cel matrix secretion. These results suggested that bone marrow mesenchymal stem cells can grow and differentiate into chondrocytes on the Bio-gide col agen membrane.

To prepare ginseng saponin Compound K with ultrasound-assisted total zymolytic ginseng saponins. The conversion rate was taken as the index to detect the pre-treatment factors such as ultrasonic power and ultrasonic time, as well as the impact of enzymatic factors, such as pH value, temperature, concentration of substrate, dosage of enzyme and reaction time, on the conversion rate. The response surface method was used to optimize the preparation conditions. The enzymolytic products were identified with MS, 1H-NMR and 13C-NMR. The results showed that the optimum conditions of the ultrasound-assisted enzymolysis were 250 W for ultrasonic power, 15 min for ultrasonic time, 5.5 for enzymolytic pH, 50 degrees C for enzymolytic temperature, 36 h for enzymolytic time, 4:5 for enzymolytic dosage: substrate and 1.0 g x L(-1) for concentration of substrate. The relative molecular mass of reaction products was 622.4. Therefore, the nuclear magnetic map verified that the reaction product was rare ginseng saponin Compound K. Under the above conditions, based on the total zymolytic ginseng saponins, the conversion rate of rare ginseng saponin Compound K was 6.91% in proportion to the total of ginsenosides. The process features gentle reaction conditions, high conversion rate and simple and reliable process, which is suitable for industrial production.
Chemistry, Pharmaceutical ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Enzymes ; chemistry ; Panax ; chemistry ; Plant Roots ; chemistry ; Saponins ; chemistry ; isolation & purification ; Ultrasonics ; methods