OBJECTIVE To investigate the in vitro inhibitory mechanism of allicin against carbapenem-resistant Klebsiella pneumoniae(CRKP).METHODS The standard microbroth dilution method was employed to determine the minimum inhibitory concentration(MIC)of allicin against the experimental CRKP.Growth curve experiments were conducted to verify the relationship between allicin concentration and antibacterial action.RNA sequencing(RNA-seq)was performed on allicin-treated CRKP and a normal control group to explore the molecular mecha-nism of allicin inhibiting CRKP growth.RESULTS The MIC of allicin against CRKP was 140 mg/ml.A total of 1 775 differentially expressed genes were screened from the normal control group and the allicin-treated group,with 845 genes upregulated and 930 genes downregulated.The expression of bacterial phosphotransferase system(PTS)genes showed significant differences between the control group and the treated group(P＜0.05),and the expression levels of EIIA-EIID genes decreased(P＜0.05).CONCLUSION Based on the analysis of RNA-seq re-sults,it is found that allicin may inhibits the growth of CRKP by disrupting its sugar transport system,thereby affecting its biofilm formation.

OBJECTIVE To investigate the in vitro inhibitory mechanism of allicin against carbapenem-resistant Klebsiella pneumoniae(CRKP).METHODS The standard microbroth dilution method was employed to determine the minimum inhibitory concentration(MIC)of allicin against the experimental CRKP.Growth curve experiments were conducted to verify the relationship between allicin concentration and antibacterial action.RNA sequencing(RNA-seq)was performed on allicin-treated CRKP and a normal control group to explore the molecular mecha-nism of allicin inhibiting CRKP growth.RESULTS The MIC of allicin against CRKP was 140 mg/ml.A total of 1 775 differentially expressed genes were screened from the normal control group and the allicin-treated group,with 845 genes upregulated and 930 genes downregulated.The expression of bacterial phosphotransferase system(PTS)genes showed significant differences between the control group and the treated group(P＜0.05),and the expression levels of EIIA-EIID genes decreased(P＜0.05).CONCLUSION Based on the analysis of RNA-seq re-sults,it is found that allicin may inhibits the growth of CRKP by disrupting its sugar transport system,thereby affecting its biofilm formation.

Oral and maxillofacial malignant tumors threaten the life and health of patients, and seriously affect their swallowing, language function and face. 125I seeds brachytherapy for oral and maxillofacial malignant tumors has been widely concerned and studied because of its advantages such as less surgical trauma, large and uniform dose distribution in the target tissue, little damage to the surrounding normal tissue, and reducing radiation exposure of medical staff. Low-dose brachytherapy with 125I seeds can effectively reduce the tumor volume and prolong the survival time of patients. This article reviews the clinical application of 125I seeds brachytherapy in oral and maxillofacial malignant tumors.

AIM: To investigate the changes of angiotensin II, its receptors and nitric oxide (NO) expression in rats with acute kidney injury (AKI) induced by endotoxin (LPS) and to assess the efficacy of different doses of dexamethasone (DXM). METHODS: Wistar rats were randomly divided into five groups as follows: control group (NC), LPS group, and DXM treatment groups of different doses of DXM (0.5, 1.0 and 5.0 mg/kg). The AKI group was injected with LPS through the lateral tail vein, and the intervention group was given different doses of DXM after LPS injection. Tissue samples were collected at 2, 6, 12 and 24 h after treatment. Serum creatinine and urea nitrogen levels were determined by automatic biochemical analyzer. H&E staining was used to observe renal histopathology. Serum TNF-α and MIP-1α levels were determined by ELISA. The expression of the angiotensin receptor 1 (AT1R) and angiotensin receptor 2 (AT2R) proteins were detected by Western blot and immunohistochemistry. Nitrate reductase was used to detect NO changes in the serum and renal tissues. RESULTS: The serum levels of serum TNF-α, MIP-1α, creatinine, and urea nitrogen were significantly increased in the LPS group compared with the control group (P<0.05). A similar trend was also observed in the levels of plasma and renal tissue AngII, renal tissue AT2R, serum and renal tissue NO (P<0.05). The expression of AT1R in renal tissue was significantly decreased in the LPS group compared with the control group (P<0.05). Pathological analysis showed that glomerular neutrophil infiltration and renal tubular epithelial cells swelling, vacuolar degeneration and necrosis in the LPS group. Prolongation LPS treatment resulted in more significant kidney damage. The serum levels of TNF-α, MIP-1α, creatinine, and urea nitrogen were significantly decreased in the DXM group compared with the LPS group (P<0.05). A similar trend was also observed in the levels of plasma and renal tissue AngII, renal tissue AT2R, serum and renal tissue NO in the DXM group (P<0.05). The expression of AT1R in renal tissue was significantly increased (P<0.05) in the DXM group compared with the LPS group, indicating the alleviation of kidney injury. Amongst these biomarkers, the levels of serum TNF-α, MIP-1α, plasma AngII showed the most significant decrease in the high dose DMX group (P<0.05). The levels of serum creatinine, urea nitrogen, kidney NO showed more significant decreases in the low and medium dose DMX groups (P<0.05). The levels of kidney AngII to dose and AT1R showed the most significant decrease in medium and high dose DXM groups (P<0.05). The levels of kidney AT2R and serum NO were not significantly different between the DXM treatment groups. CONCLUSION: LPS can induce AKI in rats that can be mitigated by DXM. The mechanism of DXM in protection against AKI may be related to the down-regulation of inflammatory factors such as AngII, AT2R, and NO, and the up-regulation of AT1R expression. Different doses of dexamethasone have different intervention effects for different effector molecules.

Objective: To explore the application of endoscopic tattooing with carbon nanoparticles in the treatment of advanced colorectal cancer (ACRC). Methods: A randomized controlled study was used. Inclusion criteria: (1) age more than 18 years old, and colorectal cancer was found for the first time and confirmed by colonoscopy and biopsy; (2) advanced colorectal cancer (preoperative TNM stage of T3/N1 or above, local unresectable lesion, M1 stage and simultaneously resectable metastatic lesion), and patients agreed to receive neoadjuvant therapy; (3) advanced colorectal cancer (TNM stage of T3/N1 or above) with simultaneous unresectable metastatic lesion, and patients refused operation and consented to chemoradiotherapy. Patients with previous abdominal surgery history, radiotherapy and chemotherapy history, urgent need for surgery or endoscopic stent placement and those with severe allergic constitution were excluded. Based on the above criteria, 120 patients diagnosed with ACRC in No.900 Hospital of the Joint Logistics Team from January 2016 to December 2017 were prospectively enrolled and randomly divided into tattoo group and non-tattoo group by random number table method. Tattoo group were tattooed within 1-7 days before chemoradiotherapy. The labeling location of the lesions: (1) if the colonoscopy could pass smoothly, 4 points were injected into the intestinal wall of the both opposite sides 1 cm cephalad and caudad of the tumor; (2) if the colorectal cavity was severely narrow and the colonoscopy could not pass, only 4 points were injected in 4 quadrants at 1 cm caudad of the tumor. Each injection point was injected with 0.1 ml carbon nanoparticles, and the size of the tumor was measured according to the range of carbon nanoparticles staining. The efficacy was evaluated after 8 weeks of chemoradiotherapy. Patients who were defined to be suitable for operation underwent operation 6 weeks after chemoradiotherapy. The following parameters were compared between two groups: lesion identification time, operation time, blood loss, distance from lesion to distal margin, the rate of first positive margin and the rate of anal sphincter preservation (rectal cancer). Among patients who had been evaluated as having no indication for surgery, those who were effective in chemoradiotherapy continued to receive chemotherapy in the original regimen; if the treatment failed, the chemotherapy regimen was replaced, and the efficacy was finally evaluated after six months [referring to the revised RECIST guidelines (version 1.1)]. Results: Three patients withdrew from this study, and 117 patients were enrolled in this study finally, including 59 cases in tattoo group and 58 cases in the non-tattoo group. There were no significant differences in baseline data between two groups (all P>0.05). All the patients had slight adverse reactions of radiotherapy and chemotherapy before operation, and could tolerate after symptomatic management without interruption of treatment. All the patients in the tattoo group had no discomfort such as fever, abdominal pain, abdominal distention, hematochezia, etc. and the intestinal mucosa could be seen clearly with black staining after being tattooed. A total of 77 patients were evaluated with surgical indications, including 39 cases in the tattoo group (tattoo-operable) and 38 cases in the non-tattoo group (non-tattoo-operatable). There were no significant differences in baseline data between the two groups (all P>0.05). Forty patients without operation indications continued chemoradiotherapy, including 20 cases in tattoo group (tattoo-inoperable) and 20 cases in non-tattoo group (non-tattoo-inoperable), whose differences in baseline data between the two groups were not significant as well (all P>0.05). No obvious edema, necrosis or abscess were found in the tattooed segments and the black spots could be seen quickly and clearly on the serosa of rectum in tattoo-operable patients. As compared to non-tattoo group, tattoo group had significantly shorter lesion identification time [(3.4±1.4) minutes vs. (11.8±3.4) minutes, t=-14.07,P<0.001], shorter operation time [(155.7±44.5) minutes vs. (177.2±30.2) minutes, t=-2.48,P=0.015], less blood loss [(101.3±36.7) ml vs.(120.2±38.2) ml, t=-2.22,P=0.029], shorter distance from lesion to distal margin [(3.7±1.0) cm vs. (4.6±1.7) cm, t=-2.20, P=0.034], while tattoo group had slightly higher rate of anal sphincter preservation [66.7%(16/24) vs. 45.5%(10/22), χ²=2.10,P=0.234] and lower rate of first positive resection margin [0 vs. 4.5%(1/22), χ²=0.62,P=0.480], but their differences were not significant. There were no significant differences in the degree of tumor differentiation and TNM stage between two groups. Patients without operative indication were evaluated for efficacy of chemoradiotherapy again after half a year. One case of complete response (CR), 8 of partial response (PR), 10 of stable disease (SD) and 1 of progressive disease (PD) were found and the improvement rate was 45.0% (9/20) in tattoo-inoperable patients. No case of CR, 6 of PR, 11 of SD and 3 of PD were found and the improvement rate was 30.0% (6/20) in non-tattoo-inoperable patients. There was no significant difference in the improvement rate between the two groups (P=0.514). Conclusions Endoscopic tattooing with carbon nanoparticles injection is safe and reliable for colorectal tumor positioning. It can assist rapid detection of lesions during surgery after neoadjuvant treatment, perform accurate resection, significantly shorten the operation time and reduce surgical trauma; can assist colonoscopy accurately to measure the size of the lesions before and after chemoradiotherapy, and increase the means of assessing the efficacy to guide the follow-up treatment plan. This technique is worth clinical promotion and application.

Objective:To investigate the expression and significance of the angiotensin Ⅱ and its receptors in lipopolysaccharide (LPS)-induced acute lung injury and acute kidney injury in rats.Methods:Forty eight Wistar rats were randomly divided into two groups: control group and endotoxin group (LPS group). LPS was injected through tail vein in LPS group, and the same amount of saline was injected through tail vein in control group.Samples were collected at 2 h, 6 h, 12 h and 24 h, respectively.The histopathology of lung and kidney was observed by HE staining.We detected lung wet/dry weight ratio, serum creatinine, urea nitrogen and Ang Ⅱ concentration in plasma, lung and kidney tissues by radioimmunoassay.Western blot and immunohistochemistry were used to detect the expression changes of AT1R and AT2R in lung and kidney tissue.Results:Compared with the control group, the pathology of lung and kidney tissue in LPS group showed different degrees of damage.The lung wet/dry weight ratio, serum creatinine and urea level in LPS group were significantly increased than that in control group( P<0.05). The Ang Ⅱ content in plasma increased significantly at 2 h and 6 h ( P<0.05), and the expression level of Ang Ⅱ in lung and kidney increased significantly at all time points ( P<0.05). The expression of AT1R in lung and kidney decreased significantly ( P<0.05), while the AT2R protein expression increased significantly ( P<0.05). Additionally the correlation analysis showed that the expression level of Ang Ⅱ and AT2R were positively correlated with lung and renal function, while the expression of AT1R was negatively correlated with lung and renal function. Conclusion:LPS results in the damage of lung and kidney function and the change of renin-angiotensin system.The changes of Ang Ⅱ and angiotension receptors were correlated with lung and kidney injury.Ang Ⅱ and angiotension receptors may be involved in LPS induced lung and kidney injury.

Objective To investigate the value of CD5 molecule-like protein(CD5L) and partial pressure of CO2 in arterial blood(PaCO2) in predicting the survival of patients with severe asthma and mechanical ventilation.Methods From Jan.2013 to Jan.2016,a total of 38 patients with severe asthma and mechanical ventilation were enrolled.Acute Physiology and Chronic Health EvaluationⅡ(APACHE II)were used to assess the severity.Enzyme linked immunosorbent assay was used to detect serum levels of CD5L at admission and 6 h after treatment.PaCO2 were also detected.Pearson correlation analysis was used to analyze the correlation between APACHEⅡ score and CD5L and PaCO2 levels.Receiver operation characteristic(ROC) curve was used to evaluate the value of CD5L and PaCO2 in predicting the survival of patients.Results APACHEⅡscores of survival patients were significantly higher than dead patients(P<0.05).CD5L level of survival patients after treatment was significantly lower than dead patients,while PaCO2 level was significantly higher(P<0.05).APACHEⅡ score was negatively correlated with serum CD5L level(r=-0.347,P<0.05),while positively correlated with PaCO2 level(r=0.573,P<0.05).ROC curve analysis showed that serum CD5L and PaCO2 were with predictive value for prediction the survival of patients,with sensitivity of 93.33%,specificity of 75.00%,accuracy of 89.47%,positive predictive value of 93.33%,and negative predictive value of 75.00% for CD5L,and those for PaCO2 were 90.00%,87.50%,89.47%,97.42% and 70.00%.Conclusion With the decreasing of CD5L level and increasing of PaCO2 level,severity of disease in patients with severe asthma and mechanical ventilation could be more serious condition,indicating poor prognosis.CD5L and PaCO2 could be with fine predictive value of survival of patients with severe asthma and mechanical ventilation.

Objective:To investigate the effects of lipopolysaccharide (LPS)on the acute lung injury (ALI)and expressions of aquaporin 1 (AQP1)and aquaporin 5 (AQP5)in lung tissue of the rats. Methods:Forty-eight SPF grade male Wistar rats were randomly divided into control group and LPS group (n=24).The rats in LPS group were intravenously injected with LPS to induce ALI models,and the rats in control group were injected with saline. The rats were sacrificed at 2,6,12 and 24 h,and the samples were collected after the successful modeling.The pathological changes of lung tissue were observed with HE staining;the lung wet/dry weight (W/D)ratio and lung permeability index were detected;ELISA was used to detect the levels of TNF-αand MIP-1α.The expression levels of AQP1 and AQP5 protein and mRNA were measured by Western blotting,immunohistochemistry and Real-Time PCR methods. Results:Compared with control group, the TNF-α and MIP-1α levels in LPS group were significantly elevated at 2,6 and 12 h (P<0.05),and at 24 h they were gradually reduced to the normal level. The HE staining results showed the alveolar and interstitial edema at 2 h after LPS injection,obviously in 12 h. The lung W/D ratios and pulmonary permeability indexes at different time points in LPS group were significantly higher than those in control group (P<0.05),and they reached the peak at 12 h.The expression levels of AQP1 and AQP5 mRNA and protein in lung tissue of the rats at different time points in LPS group were significantly lower than those in control group (P<0.01 ). Conclusion:LPS can induce ALI in the rats and down-regulate the expressions of AQP1 and AQP5;LPS is involved in the formation of pulmonary edema.

Objective To develop an ultra?high performance liquid chromatography mass spectrometer method for the rapid determination of 8?hydroxy?2'?deoxyguanosine (8?OHdG) in human urines. Methods 8?OHdG standard solution with the concentration from 0.01 to 0.1μg/ml was formulated. The solution was implanted into ion source with a rate of 7μl/min, the mass?to?charge ratio of parent ion and product ions, and ion mass to charge ratio was identified. The mass spectrum parameters of each Ion pairs, such as DP, EP and EXP, were gradually optimized. The urine sample with a concentration of 10.0μg/L was detected, and the pH of the sample was adjusted using 1 mol/L ammonium formate and formic acid solution with a volume ratio of 5∶1, 4∶1, 3∶1, 2∶1, and 1∶1. It was tested using three different polarity of SPE, i.e.:HLB, MCX, and MAX. The elution effect of methanol and water mixture with the proportion of 90∶10, 80∶20, 50:50, 20:80, and 10:90 were tested, and then acetonitrile and water mixture with the proportion of 90∶10, 80∶20, 50∶50, 20∶80, 10∶90 were also tested. The standard curve was constructed using the ratio of a standard series application fluid concentration to corresponding compounds quantitative ion liquid concentration of the peak area. The detection limit was determined as 3 times of the signal to noise ratio corresponding to the concentration of 8?OHdG, and the quantitative lower limit was determined as 10 times of the signal to noise ratio corresponding to the concentration of 8?OHdG. The blank urine spiked recovery method was used to evaluate the precision and recovery rate. Results The mass to charge ratio of parent ion was 284.1 and the product ions was 168.1, 140.1, 123.0, and 112.0, respectively. The collision voltage of quantitative ion?pair 284.1/168.1 was 18 V, the 284.1/140.1 collision voltage was 42 V, the 284.1/123.0 collision voltage was 48 V, and the 284.1/112.0 collision voltage was 53 V. The recovery rate was the highest (87.9%-104.3%) when the pH of urine was adjusted by a 10 ml 1 mol/L ammonium formate solution, 2 ml of formic acid, 88 ml of water are mixed with the sample solution volume ratio of 1∶5, and then purified with 3 ml of methanol and 3 ml water activated HLB extraction column. Within 1.0-100.0 μg/L concentration range, 8?OHdG standard application solution test results showed a good linear relationship. The regression equation was y=1.25x+0.74, and the correlation coefficient was r=0.999 5. The detection limit was 0.2μg/L, and the limit of quantification was 0.7μg/L. The method of recovery rate was in the range of 87.9%to 104.3%, the precision was in the range from 1.5% to 3.7% and inter?assay precision was in the range from 1.6% to 5.4%. Conclusion The method developed in this study had high sensitivity, good precision and accuracy, and a wide range of testing concentrations.

Objective To develop an ultra?high performance liquid chromatography mass spectrometer method for the rapid determination of 8?hydroxy?2'?deoxyguanosine (8?OHdG) in human urines. Methods 8?OHdG standard solution with the concentration from 0.01 to 0.1μg/ml was formulated. The solution was implanted into ion source with a rate of 7μl/min, the mass?to?charge ratio of parent ion and product ions, and ion mass to charge ratio was identified. The mass spectrum parameters of each Ion pairs, such as DP, EP and EXP, were gradually optimized. The urine sample with a concentration of 10.0μg/L was detected, and the pH of the sample was adjusted using 1 mol/L ammonium formate and formic acid solution with a volume ratio of 5∶1, 4∶1, 3∶1, 2∶1, and 1∶1. It was tested using three different polarity of SPE, i.e.:HLB, MCX, and MAX. The elution effect of methanol and water mixture with the proportion of 90∶10, 80∶20, 50:50, 20:80, and 10:90 were tested, and then acetonitrile and water mixture with the proportion of 90∶10, 80∶20, 50∶50, 20∶80, 10∶90 were also tested. The standard curve was constructed using the ratio of a standard series application fluid concentration to corresponding compounds quantitative ion liquid concentration of the peak area. The detection limit was determined as 3 times of the signal to noise ratio corresponding to the concentration of 8?OHdG, and the quantitative lower limit was determined as 10 times of the signal to noise ratio corresponding to the concentration of 8?OHdG. The blank urine spiked recovery method was used to evaluate the precision and recovery rate. Results The mass to charge ratio of parent ion was 284.1 and the product ions was 168.1, 140.1, 123.0, and 112.0, respectively. The collision voltage of quantitative ion?pair 284.1/168.1 was 18 V, the 284.1/140.1 collision voltage was 42 V, the 284.1/123.0 collision voltage was 48 V, and the 284.1/112.0 collision voltage was 53 V. The recovery rate was the highest (87.9%-104.3%) when the pH of urine was adjusted by a 10 ml 1 mol/L ammonium formate solution, 2 ml of formic acid, 88 ml of water are mixed with the sample solution volume ratio of 1∶5, and then purified with 3 ml of methanol and 3 ml water activated HLB extraction column. Within 1.0-100.0 μg/L concentration range, 8?OHdG standard application solution test results showed a good linear relationship. The regression equation was y=1.25x+0.74, and the correlation coefficient was r=0.999 5. The detection limit was 0.2μg/L, and the limit of quantification was 0.7μg/L. The method of recovery rate was in the range of 87.9%to 104.3%, the precision was in the range from 1.5% to 3.7% and inter?assay precision was in the range from 1.6% to 5.4%. Conclusion The method developed in this study had high sensitivity, good precision and accuracy, and a wide range of testing concentrations.