0.05).The average fading time of the lymph nodes stained by PB and MB+PB was 13.06min and 82.13min,we observed the lymph nodes stained by MB 120 min,they were still stained deeply.There were dramatically differences among three groups(P=0.000).Conclusion Application of MB and PB possess their advantage and disadvantage in sentinel node biopsy.The mixture of patent blue and methylene blue could overcome the shortage of the two vital dyes in the experimental study and be an ideal tracer for the use.

Objective The volatile oil was further purified to get the patchouli alcohol by molecular distillation taking patchouli oil which contained 23.97% of patchouli alcohol as raw material.Methods The best process parameters,including temperature,feed rate,and scraping rate,of molecular distillation technique for separating patchouli oil were studied by orthogonal test.The content was detected by GC under the conditions of elastic quartz capillary column HP-5,detector temperature 280 ℃,gasification temperature 280 ℃,distributary refer 20∶1.The effects of evaporation temperature on content and extraction rate of the patchouli oil had been investigated and the difference between the molecular and vapor distillations was studied.Results Content and extraction rate of patchouli alcohol under the best evaporation temperature 65 ℃ by molecular distillation reached to 40.71% and 76.55%,respectively.Conclusion The molecular distillation technique that could get the product with higher purity and extracting rate than that by vapor distillation in patchouli oil purification has a very good prospect in industry.

Although there is emerging evidence that BJ46a can function as potent inhibitor of the SVMPs proteolytic activities,its anticancer effect on invasion and metastasis has not yet been evaluated.Inhibition effect of BJ46a on experimental pulmonary metastasis in mice inoculated with B16 melanoma cells at the protein level was investigated. First,BJ46a was produced in baculovirus expression system. SDS-PAGE and Western blot analysis confirmed that BJ46a recombinant protein was produced by Sf9 cells infected with high-titer viral stock. Then,recombinant fusion protein was purified by ProBondTM at the point of maximal expression. B16 cells pre-treated with recombinant BJ46a injected into C57BL/6 mice via the tail lateral vein to form experimental pulmonary metastasis model. The numbers of metastatic lesions in C57BL/6 mice changed dramatically:BJ46a different concentrations of recombinant protein group were 1.1 ? 0.83,0.9 ? 0.7,significantly lower than the control group (6.3?3.00,P

Objective:To investigate the changes of sTim-3,HMGB1 and TGF-β in the brucellosis patients and to analyse the relationship between the changes of these molecules and brucella infection. Methods:28 cases of brucellosis patient untreated and 28 healthy control cases in the age and gender matched with brucellosis cases were collected. The serum levels of sTim-3 and HMGB1 were detected by ELISA,and the levels of Spot forming cells secreting TGF-β were measured by ELISPOT in patients and healthy control group. Results: Compared with healthy controls, sTim-3/HMGB1 expression levels and Spot forming cells secreting TGF-β were significantly increased in the brucellosis patients ( P<0. 01 ) . The changes of Spot forming cells secreting TGF-β were positively correlated with the levels of HMGB1 (P<0. 05). Conclusion:The serum levels of sTim-3/HMGB1 and Spot forming cells of secreting TGF-β from peripheral blood mononuclear cell are significantly increased in the brucellosis patients. Those molecules may be involved in the process of brucella infection and may play a significant role in the immune escape of patients infected with brucella.

Objective: This study was performed to analyze the feasibility of elective nodal irradiation (ENI) for early-stage esophageal carcinoma (EEC), evaluate the curative effect of radiation therapy for EEC, and determine the causes of treatment failure. Methods:Data were collected from 123 patients with esophageal squamous cell carcinoma of clinical T1-2 N0-1 M0 stage. Patients were divided into two groups based on different types of radiation therapy. Among the 123 patients, 102 underwent involved field irradiation (IFI), whereas 21 received ENI. A comparative analysis of patients in the two groups was conducted. Results:In 123 patients, the one-, three-, and five-year overall survival rates were 87.8%, 47.2%, and 36.6%, respectively. By contrast, the one-, three-, and five-year local control rates were 89.4%, 67.5%, and 48.8%, respectively. After comparing the one-, three-, and five-year survival rates in the ENI patients (i.e., 90.5%, 47.6%, and 42.9%, respectively) with those in the IFI patients (i.e., 86.3%, 49.0%, and 35.2%, respectively), no significant difference was found (χ2=0.290, P=0.588 8). The results indicate that ENI possibly decreased nodal metastases (χ2=5.778, P=0.016). Conclusion:Three-dimensional conformal radiotherapy is one of the best therapeutic regimens of radiation for EEC. ENI is possibly effective for preventing regional nodal metastasis. Whether ENI leads to an improved overall survival needs further investigation.

OBJECTIVE: To investigate whether dynamic monitoring of citrulline (Cit) has guiding value for early enteral nutrition (EN) in patients with severe gastrointestinal injury. METHODS: A observational study was conducted. A total of 76 patients with severe gastrointestinal injury admitted to different intensive care units of Suzhou Hospital Affiliated to Nanjing Medical University from February 2021 to June 2022 were enrolled. Early EN was performed in 24-48 hours after admission as recommended by the guidelines. Those who did not terminate EN after 7 days were enrolled in the early EN success group, and those who terminated EN within 7 days due to persistent feeding intolerance or deterioration of general condition were enrolled in the early EN failure group. There was no intervention during the treatment. Serum Cit levels were measured by mass spectrometry at admission, before EN starting and EN 24 hours, respectively, and the changes in Cit within EN 24 hours (ΔCit) were calculated (ΔCit = EN 24-hour Cit-Cit before EN starting). Receiver operator characteristic curve (ROC curve) was plotted to investigate the predictive value of ΔCit for early EN failure, and the optimal predictive value was calculated. Multivariate unconditional Logistic regression was used to analyze the independent risk factors for early EN failure and death at 28 days. RESULTS: Seventy-six patients were enrolled in the final analysis, of which 40 succeeded in early EN and 36 failed. There were significant differences in age, main diagnosis, acute physiology and chronic health evaluation II (APACHE II) score at admission, blood lactic acid (Lac) before EN initiation and ΔCit between the two groups. Multivariate Logistic regression analysis showed that age [odds ratio (OR) = 0.929, 95% confidence interval (95%CI) was 0.874-0.988, P = 0.018], ΔCit (OR = 2.026, 95%CI was 1.322-3.114, P = 0.001) and increased feeding rate within 48 hours (OR = 13.719, 95%CI was 1.795-104.851, P = 0.012) were independent risk factors for early EN failure in patients with severe gastrointestinal injury. ROC curve analysis showed that ΔCit had a good predictive value for early EN failure in patients with severe gastrointestinal injury [area under the ROC curve (AUC) = 0.787, 95%CI was 0.686-0.887, P < 0.001], and the optimal predictive value of ΔCit was 0.74 μmol/L (sensitivity was 65.0%, specificity was 75.0%). Combined with the optimal predictive value of ΔCit, "overfeeding" was defined as ΔCit < 0.74 μmol/L and increased feeding within 48 hours. Multivariate Logistic regression analysis showed that age (OR = 0.825, 95%CI was 0.732-0.930, P = 0.002), APACHE II score (OR = 0.696, 95%CI was 0.518-0.936, P = 0.017) and early EN failure (OR = 181.803, 95%CI was 3.916-8 439.606, P = 0.008) were independent risk factors for 28-day death in patients with severe gastrointestinal injury. The new variable "overfeeding" was also associated with an increased risk of death at 28 days (OR = 27.816, 95%CI was 1.023-755.996, P = 0.048). CONCLUSIONS Dynamic monitoring of Cit has guiding value for early EN in patients with severe gastrointestinal injury.
Humans ; Infant, Newborn ; Enteral Nutrition ; Citrulline ; APACHE ; Abdominal Injuries ; Cognition ; Thoracic Injuries

Objective To identify the influencing factors for rural doctors′ training effect,and suggest on the improvement of such training.Methods On the basis of rural doctors′ survey,the theory of pre-intervention was used to probe into the influencing factors for such training in five dimensions of attention notice,mega-cognitive strategies,advance organizer,goal orientation,and preparatory information.Results 73.2% of the groups were found with satisfying effect.In the single factor analysis,comparison of training effect involving such factors as age,gender,length of work life and pre-intervention revealed statistical significance(P<0.05).As shown in the logistic regression analysis,High motivation in meta-cognitive strategies,Clear goals in goal orientation,Tiered and categorized training in advance organizers,and Practical learning in preparatory information,as well as variants like age would influence rural doctors′ training effect significantly.Conclusions The key to better training effect lies in better motivation of the trainee,setting correct training goals,emphasis on the practicability of training contents and,the pertinence of the training objects.

AIM: To construct pVAX1-GrB. METHODS: Lymphocytes from human laryngeal carcinoma tissue were separated from tumor tissue. The fragment of granzyme B (GrB) was amplified by RT-PCR and was recombined to the downstream of T7 promoter in the vector pVAX1. The construction was transfected into Hep2 cells with lipofectamine 2000. The expression of protein was identified by indirect immunofluorescent antibody assay. RESULTS: It has been proved that the sequence of the RT-PCR product was totally consistent with the data of GenBank by DNA sequencing analysis. The GrB cDNA fragment was cloned into the vector of pVAX1 in the right direction and the open reading fragment of GrB was maintained. The target protein was detected in the transfected Hep2 cells. CONCLUSION: The pVAX1-GrB plasmid was successfully constructed and expressed. [

Objective:To construct GJB2 gene mutaitons common in Chinese EGFP fusion protein vectors, and to search for better way to study the mechanism of deletion mutaitons in GJB2 gene. Method: Non-fusion protein vectors of 235delC, 299-300 del AT and 176 del 16 bp were first made by point mutaiton methods in vitro. Then expression part of the upper 3 mutations were amplified by PCR and the PCR products were cloned into TA cloning vector. After cutting by restriction enzymes EcoRI/BamHI, three deletion mutaions were inserted into pEG-FP-N1 vector. Sequencing was used to verify the validity of the fusion protein vectors. HEK293 cells were trans-fected with the recombinant DNA samples by the liposome complex method. Results The recombined plasmids were highly expressed in HEK293 cells. Green fluorescence singals were distributed uniformly in cytoplasm. Conclusion; GJB2 mutations common in Chinese EGFP fusion protein vectors were constructed successfully. It may provide a better way to explore the reasons of nonsyndromic hearing loss common in Chinese.

OBJECTIVETo investigate the potential effects of angiogenic process by secretory phospholipase A2 (sPLA2) inhibitor-HyPE (linking N-derivatized phosphatidyl-ethanolamine to hyaluronic acid) on human bone marrow endothelial cell line (HBME-1).

METHODSIn order to examine the suppressing effects of HyPE on HBME-1 proliferation, migration, and capillary-like tube formation, HBME-1 were activated hy angiogenic factor, specifically by basic fibroblast growth factor (b-FGF), vascular endothelial growth factor (VEGF), and oncostatin M (OSM) (at a final concentration of 25, 20, and 2.5 ng/mL, respectively), then HBME-1 proliferation, migration, and tube formation were studied in the absence or presence of HyPE. HBME-1 tube formation was specially analyzed in fibrin gel.

RESULTSHyPE effectively inhibited HBME-1 proliferation and migration as a dose-dependent manner, whatever HBME-1 were grown in the control culture medium or stimulated with b-FGF, VEGF, or OSM. In fibrin, the formations of HBME-1 derived tube-like structures were enhanced by all angiogenic factors, but these were strongly suppressed by HyPE.

CONCLUSIONSThe results support the involvement of sPLA2 in angiogenesis. It is proposed that sPLA2 inhibitor introduces a novel approach in the control of cancer development.

Bone Marrow Cells ; cytology ; Capillaries ; drug effects ; Cell Division ; drug effects ; Cell Movement ; drug effects ; Cells, Cultured ; Endothelial Cells ; cytology ; Enzyme Inhibitors ; pharmacology ; Fibroblast Growth Factor 2 ; antagonists & inhibitors ; Humans ; Hyaluronic Acid ; pharmacology ; Neovascularization, Pathologic ; pathology ; Oncostatin M ; Peptides ; antagonists & inhibitors ; Phosphatidylethanolamines ; pharmacology ; Phospholipases A ; antagonists & inhibitors ; Phospholipases A2 ; Vascular Endothelial Growth Factor A ; antagonists & inhibitors