OBJECTIVE: To establish a high-performance liquid chromatographic(HPLC)-fluorometric method for the concentration determination of itraconazole (ITZ) and its metabolite hydroxy-itraconazole(HITZ) in rabbit plasma simultaneously.METHODS: Separation was achieved with Agilent Eclipse C18,the mobile phase consisted of acetonitrile(adjusted to pH=2.5 with 85% phosphoric acid)-water(63∶37),and the flow rate was 1.0 mL?min-1.The excitation and emission wavelengths were 260 nm and 365 nm.The analysis was performed at room temperature..RESULTS: The linear concentration range of ITZ and HITZ were 7.4～296 0 ng?mL-1(r=0.999 1)and 5.4～216 0 ng?mL-1(r=0.999 3), respectively.The limit of detection was 0.518 and 0.318 ng?mL-1,respectively.The extraction and method recovery rate of ITZ and HITZ were more than 80%.The intra-day and inter-day RSD of ITZ and HITZ were less than 4.47%. CONCLUSIONS: The method is simple,sensitive,accurate and reliable for monitoring ITZ and HITZ in rabbit plasma and pharmacokinetic studies.

Objective To observe the effects of road transport on hematological and biochemical parameters in New Zealand rabbits.Methods A total of 12 healthy New Zealand rabbits were selected for 2 h road transport.Blood samples were collected at 0, 24, 48, 72 and 96 h after transport, respectively.White blood cells (WBC), red blood cells (RBC), hemoglobin (HGB), hematocrit (MCV), mean erythrocyte hemoglobin content (MCH), mean erythrocyte hemoglobin concentration (MCHC) and platelets (PLT) were measured using a blood analyzer.Blood alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin (ALB), total protein (TP), urea nitrogen (UREA), creatinine, uric acid (UA), triglycerides (TG), total cholesterol (COHL), glucose (GLU), hypersensitive C-reactive protein (CRP), α-amylase (AMYL), and creatine kinase (CK) were detected by an automatic biochemical analyzer.Results Compared the parameters before and after transport, The WBC count was increased first (P< 0.05 or P < 0.01) and then decreased after transport, the levels of RBC, HGB, HCT and PLT were decreased first (P< 0.05 or P < 0.01) and then increased after transport, and MCV was significantly high at 96 h after transport (P< 0.05).Among the clinical biochemical parameters, ALT, AST and BUN were firstly elevated (P< 0.05 or P < 0.01) and then decreased.TP, ALB as well as CREA and TG were firstly decreased (P< 0.05 or P < 0.01) and then increased.GLU was significantly low at 24 h after transport (P< 0.05).All parameters except MCV at 96 h after transport were not significantly different from those before transport.Conclusions Changes of blood routine, liver and kidney function indexes, lipid metabolism indexes, glucose metabolism index and creatine kinase index are observed in the New Zealand rabbits after 2-hour road transportation, and all the indicators except MCV return to pre-transport levels within 96 h.

Objective To explore whether lymphovascular invasion can be used as a prognostic indicator in patients with bladder cancer underwent radical cystectomy.Methods The series included clinical data of 459 patients who had underwent radical cystectomy for bladder cancer between January 2006 and December 2012 in our hospital.Among all the patients,391 were male while 68 were female.Pathological grade G1 was diagnosed in 47,G2 in 104 and G3 in 308.Amount of pathological stage T1 or Tis or Ta or T0 was 167,T2 127,T3 89 and T4 76.Of all the patients,92 had lymph node metastasis.Follow-up time was between 13 and 99 months.The presence or absence of lymphovascular invasion was determined by HE staining in the radical cystectomy specimen.The x2 test was used to detect the association between lymphovascular invasion and several clinicopathological features,the Kaplan-Meier method was used to compare recurrence-free survival according to findings of lymphovascular invasion in the surgical specimen,and the multivariate Cox proportional-hazards regression model was used to assess the prognostic significance of some factors.Results Lymphovascular invasion was detected in 128 (28％) specimens.Among them,44 (34％) had tumor recurrence.Lymphovascular invasion was significantly associated with gender,tumor grade,pathological stage,lymph node metastasis and disease recurrence.Recurrence-free survival in patients without lymphovascular invasion was significantly higher than that in those with lymphovascular invasion (P＜ 0.05).Cox proportional hazards model showed that age,pathological stage and lymph node metastasis were independent predictors for disease recurrence.Conclusion In patients with bladder cancer underwent radical cystectomy,lymphovascular invasion may have a significant association with some prognostic parameters,but it can not be used as an independent predictor of disease recurrence.

Objective To identify the role of phosphatidylinositol-3-kinase(PI3K) in mediating necroptosis induced by tumor necrosis factor alpha (TNFα) and the involved mechanism.Methods Knockdown of p110α,receptor-interacting protein 1(RIP1) or both p110αand RIP1 was mediated by the specific short hairpin RNA (shRNA) lentivirus and verified by RT-PCR or Western blotting .In addition , Western blotting was used to detect phosphorylation of mixed lineage kinase domain-like protein(MLKL) and protein kinase B(AKT) or tetramerization of MLKL.Cell death was measured by micros-copy and flow cytometry.Results AKT phosphorylation and TNFα-induced necroptosis of L929 cells were suppressed by the inhibitors of PI3K or AKT, as well as p110αknockdown.Moreover, RIP1 knockdown did not inhibit L929 cell death induced by TNFαplus Z-VAD, but the RIP1-independent necroptosis was inhibited by p 110αknockdown.In addition, p110αknockdown suppressed MLKL phosphorylation and tetramerization induced by TNFαwith Z-VAD in L929 cells. Conclusion PI3K mediates necroptosis of L929 cells induced by TNFαby activating AKT and MLKL, respectively.

To get the mature peptide of human-derived neurotrophin-6 (NT-6), NT-6 gene encoding mature peptide was amplified by PCR, using the NT-6 cDNA that had been cloned as templet. The gene encoding mature peptide of NT-6 gene was cloned into pGEX1-lambdaT plasmid to construct the fusion expression vector. Expression of fusion protein in Escherichia coli was defected after induction by isopropyl beta-D-thiogalactoside(IPTG). The mature peptide of NT-6 was collected with GST fusion protein purifying kit. It was shown that a fragment of 460bp was gained by PCR. With the techniques of double-cleave and electrophoresis, the recombinant vector was identified as pGEX1-NT-6. The recombinant vector pGEX1-NT-6 transformed Escherichia coli expressed fusion protein of 41KD after induction by IPTG. Cleaved by thrombin, the mature peptide of NT-6 was obtained; its molecular weight was about 15KD. The cloning and expression of human-derived NT-6 gene encoding mature protein has provided a basis for further studies on the function and clinical application of NT-6.
Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Humans ; Isopropyl Thiogalactoside ; pharmacology ; Nerve Growth Factors ; biosynthesis ; genetics ; Peptides ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; isolation & purification

Objective:To analyze the problems existing in the management mode of clinical research coordinator (CRC) in the new context, propose improvement measures and analyze the effectiveness of improvement.Methods:Four types of stakeholders related to CRC, including investigators, clinical research associates(CRA), institutional managers and human subjects were investigated in regular monthly questionnaire survey on the basis of CRC management comparasionat home and abroad. The new CRC management countermeasures implemented by our institution in July 2020 were taken as the time node, and the data from January to June 2020 were taken as the pre-implementation group, and the data from July to December 2020 were taken as the post-implementation group. Compare the monthly scores of CRCs′ work effect for 30 projects in the institution before and after the implementation of such countermeasures.Results:The scores of CRCs′ working effectiveness were improved after the implementation of CRC management countermeasures, whcih including standardizing the entry of CRC, updating of the training and assessment mechanisms, conducting regular communication meetings and developing reward and punishment measures. The scores from investigators increased by 20.17%, scores from CRAs increased by 11.54%, scores from institutional managers increased by 14.26%, scores from subjects increased by 10.64%, and the total scores increased by 14.13%( P<0.01). Conclusions:Countermeasures to optimize CRC management taken by drug clinical trial institutions can significantly improve CRCs′ working effectiveness in multiple dimensions.

Objective: To evaluate the role of spinal COX-1 and COX-2 in remifentanil-induced hyperalgesia in mice with incisional pain. Methods: Thirty-two male C57BL/6J mice, aged 8-10 weeks, weighing 20-25 g, were divided into 4 groups (n=8 each) using a random number table method: control group (group C), incisional pain plus remifentanil group (group IR), incisional pain plus remifentanil plus selective COX-1 inhibitor group (group IR+ SC560), and incisional pain plus remifentanil plus selective COX-2 inhibitor group (group IR+ SC236). In IR, IR+ SC560 and IR+ SC236 groups, normal saline 10 μl, SC560 25 μg and SC236 25 μg were intrathecally injected, respectively, 15 min later remifentanil 10 μg/kg was injected via the tail vein for 4 times at 15 min intervals.An incisional pain model was established after the first injection of remifentanil.The mechanical paw withdrawal threshold (MWT) was measured at 24 h before normal saline or remifentanil injection and 3, 6, 24 and 48 h after the last injection (T₀-T₄). The mice were sacrificed after the last measurement of pain threshold, and the L_4-6 segments of the spinal cord were removed for determination of the expression of COX-1 and COX-2 (by Western blot) and expression of COX-1 and COX-2 mRNA (by quantitative real-time polymerase chain reaction). Results: Compared with group C, the MWT was significantly decreased, and the expression of COX-2 protein and mRNA was up-regulated in IR, IR+ SC560 and IR+ SC236 groups (P<0.05). Compared with group IR, the MWT was significantly increased in IR+ SC560 and IR+ SC236 groups (P<0.05). There was no significant difference in the MWT at each time point between IR+ SC560 and IR+ SC236 (P>0.05) .There was no significant difference in the expression of COX-2 protein and mRNA among group IR, group IR+ SC560 and group IR+ SC236 (P>0.05). There was no significant difference in the expression of COX-1 protein and mRNA among the four groups (P>0.05). Conclusion Compared with COX-1, spinal COX-2 plays a major role in the pathophysiological mechanism of remifentanil-induced hyperalgesia in mice with incisional pain.

Purpose: We aimed to explore whether the prognosis of patients treated with capecitabine and oxaliplatin (XELOX) or S-1 and oxaliplatin (SOX) regimens who received fewer cycles of chemotherapy after D2 radical resection for gastric cancer (GC) would be non-inferior to that of patients who received the standard number of cycles of chemotherapy. Materials and Methods: Data on patients who received XELOX or SOX chemotherapy after undergoing D2 radical resection at Harbin Medical University Cancer Hospital between January 2011 and May 2016 were collected. Results: In patients who received 4, 6, and 8 cycles of chemotherapy, the 5-year overall survival (OS) rates were 59.4%, 64.8%, and 62.7%, respectively. Compared to patients who received 4 cycles of chemotherapy, those who received 6 cycles (hazard ratio [HR], 0.882; 95% confidence interval [CI], 0.599–1.299; P=0.52) or 8 cycles (HR, 0.882; 95% CI, 0.533–1.458; P=0.62) of chemotherapy did not exhibit significantly prolonged OS. The 3-year disease-free survival (DFS) rate of patients who received 4, 6, and 8 cycles of chemotherapy was 62.1%, 67.2%, and 60.8%, respectively. Compared to patients who received 4 cycles of chemotherapy, those who received 6 cycles (HR, 0.835; 95% CI, 0.572–1.221; P=0.35) or 8 cycles (HR, 0.972; 95% CI, 0.606–1.558; P=0.91) of chemotherapy did not show significantly prolonged DFS. However, the 3-year DFS and 5-year OS rates of patients who received 6 cycles of chemotherapy appeared to be superior to those of patients who received 4 and 8 cycles of chemotherapy. Conclusions For patients with stage III GC, 4 to 6 cycles of XELOX or SOX chemotherapy may be a favorable option. This study provides a rationale for further randomized clinical trials.

Objective To evaluate the role of clinical pharmacists on the pharmacological monitoring and management of diabetic patients. Methods 406 adult outpatients with diabetes in outpatient were selected as research object. The patients were given the questionnaire and intervened with diabetes education and management by the clinical pharmacist regularly. The patient’s knowledge of the diabetes medication before and after intervention, blood glucose and glycosylated hemoglobin values, treatment compliance, non-reserved outpatient visit, emergency, hospitalization, etc. were compared and statistically analyzed. Results After pharmacy intervention, the patients' knowledge of diabetes and drug-related information, treatment compliance, blood glucose and glycosylated hemoglobin were better than before intervention, P<0.01. Non-reserved outpatient visits and emergency cases were better than before intervention, P<0.05. There are significant differences. Conclusion Clinical pharmacists carry out diabetes chronic disease management and build a clinical pharmacist-led chronic disease management model, which helps to promote standardized treatment, improve patient compliance, promote rationalized medication, achieve the goal of controlling blood sugar and reduce complications.

In this study, a new Bacillus velezensis strain Bv-303 was identified and its biocontrol effect against rice bacterial-blight (BB) disease caused by Xanthomonas oryzae pv. oryzae (Xoo) was investigated. Cell-free supernatant (CFS) of strain Bv-303 under different growth conditions were prepared to test the antagonistic activity and stability against Xoo by the Oxford-cup method in vitro. The antibacterial effect of strain Bv-303 to BB disease in rice were further analyzed in vivo by spraying the cell-culture broth (CCB), CFS and cell-suspension water (CSW), respectively, on the rice leaves inoculated with Xoo. Additionally, rice seeds germination rate and seedling growth under the strain Bv-303 CCB treatment were tested. The results showed that the strain Bv-303 CFS significantly inhibited Xoo growth by 85.7%‒88.0% in vitro, which was also stable under extreme environment conditions such as heat, acid, alkali and ultraviolet light. As tested in vivo, spraying the CCB, CFS or CSW of strain Bv-303 on the Xoo-infected leaves enhanced rice plant resistance to BB disease, with CCB showing the highest increase (62.7%) in disease-resistance. Notably, CCB does not have negative effects on rice seed germination and seedling growth. Therefore, strain Bv-303 has great potential for biocontrol of the rice BB disease.
Oryza ; Fatigue Syndrome, Chronic ; Bacillus ; Xanthomonas ; Plant Diseases/microbiology*