Objective To develop an ion-pairing reversed-phase high performance liquid chromatography(IP-RP-HPLC)method for the determination of L-methionine sulphoximine(MSX) residues in recombinant protein therapeutics, and to verify and preliminarily apply it, so as to provide a reference for the quality control of related products.Methods A chromatographic analysis method was screened and the mobile phase conditions such as ion-pairing reagent, pH and salt concentration were optimized to develop an IP-RP-HPLC method for the determination of MSX residues in recombinant protein therapeutics. The method was verified for specificity, system applicability, linearity, limit of detection(LOD) and limit of quantitation(LOQ), precision, accuracy and solution stability, by which the residual MSX contents in multiple batches of in-process sample and bulk of recombinant protein therapeutics were determined.Results The chromatographic conditions were determined as follows: using Hypersil GOLDTMC18 column(4. 6 mm × 250 mm, 5 μm) at column temperature of 25 ℃; 50 mmol/L sodium dihydrogen phosphate(containing 5 mmol/L sodium 1-octane sulfonate, pH 2. 0)-acetonitrile(volume fraction of 85 ∶ 15) as mobile phase for isocratic elution at a flow rate of 0. 6 mL/min; detection wavelength of 200 nm; injection volume of 50 μL. The method exhibited good specificity and system applicability. The control substance showed good linearity in the concentration range of 10-100 μg/mL, with a correlation coefficient(R) of 0. 999 95. The LOD and LOQ were 5 and 0. 5 μg/mL, respectively. MSX was not detected in either recombinant protein A or B, and the relative standard deviations(RSDs) of the measured concentrations of six parallel spiked samples were 1. 3% and 1. 1%, respectively, with the recovery rates ranged from 94% to 103%. For recombinant protein A spiked test sample placed at 10 ℃, the RSD of the measured concentrations at 0 h and 12 h were 2. 1%. MSX was not detected in the in-process samples and bulk of recombinant protein A test samples and three batches of recombinant protein B samples.Conclusion The developed IP-RP-HPLC method has the characteristics of simple operation and high sensitivity, and can be used for the quality control of MSX residues in recombinant protein therapeutics.

Objective To investigate the clinical efficacy of rectal resection for rectal cancer.Methods 35 patients with rectal cancer were diagnosed by pathological examination.They were treated with laparoscopic rectal cancer resection and conventional chemotherapy.The patients were followed up for 3 years.The operative time,blood loss,intraoperative and postoperative complications,postoperative recovery,death and so on were observed.Results The patients were operated successfully,and no tumor cells were found in the bowel edge.The average operative time was (171.74 ± 58.24) min,average blood loss was (85.74 ± 68.32) ml,there were no infection,bleeding,anastomotic complications.After 2 ～ 3 years of follow-up,there was 1 patient with liver metastases,and no local recurrence,no fecal incontinence and no deaths.Conclusion Rectal resection for rectal cancer had good effect and could improve patients' quality of life.

Objective To evaluate the clinical significance of serum tumor supplied group of factor (TSGF) in diagnosing colorectal carcinoma. Methods The TSGF concentration in the patients with colorectal carcinoma, with colorectal polyp and healthy subjects were retrospectively reviewed. Results The TSGF concentration in the colorectal carcinoma patients was significantly higher than that in the colorectal polyp patients (P

Objective To evaluate the relationship between the expression of EGF, EGFR and PCNA with the clinicopathological characteristics in human colorectal carcinoma. Methods Immunohistochemistry was used to examine the expression of EGF, EGFR and PCNA in 50 specimens of colorectal carcinoma and 10 specimens of normal colorectal tissue. Results In cancer specimens,the expression of EGF and EGFR were 76.07% and 84.15% respectively, but in normal colorectal tissue were no expression of EGF and EGFR (P

[Objective]To investigate the methods of preparing acellular cartilaginous matrix(ACM) and constructing the tissue engineered cartilage with adipose-derived stem cells(ADSCs)-seeded acellular cartilaginous matrix in vitro.[Method]The ADSCs were isolated from adult New Zealand albino rabbits by collagenase,cultured and amplified in vitro.Fresh cartilage isolated from adult New Zealand albino rabbits were freeze-dried for twelve hours and then treated with Triton X-100,Dnase and RNase so as to obtain the ACM.After sterilized with ultra-violet,the ADSCs were seeded in the acellular cartilaginous matrix at a final density of 2?107/L and cultured in chondrogenic differentiation medium for two weeks in order to construct tissue engineered cartilage.Histology,immunohistochemistry and transmission electron microscope(TEM) were applied to examine the fresh ACM and tissue engineered cartilage.[Result]The test of hematoxylin-eosin(HE) staining and TEM showed no cellular structure in the ACM with only recesses left by removed cells.The ACM had suitable interval porosity and aperture size.After integrated with ADSCs,cells migrated into the ACM and adhered to the surface of material and grew well.After cultured in chondrogenic differentiation medium for two weeks,immunohistochemical staining of type Ⅱ collagen showed part of the cells in the material had enhanced expression of type Ⅱ collagen.[Conclusion]ACM can be used as scaffold material in cartilage tissue engineering.If it was seeded with ADSCs and cultured in chondrogenic differentiation medium,ACM can be used to construct cartilage tissue successfully.

1 cm. Postoperative pathological findings showed negative cut surface. No hemorrhage or bile leakage were seen after operation. The abdominal drainage tube was indwelled for 3~4 d. The postoperative hospital stay was 3~5 d. Follow-up checkups for 1~20 months (mean, 12.5 months) revealed no tumor recurrence or incision implantation. All the patients survived. Conclusions Laparoscopic regular hepatic left lateral lobectomy can be carried out safely and effectively in clinically selected patients.

Recent evidence suggests that, in spite of the redundancy of angiogenic factors involved in pathological angiogenesis , strategies aimed at inhibiting specific endothelial cell angiogenic factors at their release or receptor level may form the basis for effective and safe treatment of angiogenic mediated disease processes. Physiologic angiogenesis is fundamental to reproduction, development and repair. Pathological angiogenesis sustains the progression of many neoplastic and proinflammatory diseases. And endothelial cell is an important target especially for the angiogenesis. The target represents a novel strategy for the treatment of the human disorders where pathological angiogenesis is involved.

Objective:To investigate the protective effects and mechanism of ligustrazine on myocardial ischemia reperfusion injury(MIRI) in rats.Methods:30 Wistar rats were divided into three groups randomly:sham operation group,ischemia reperfusion injury group(IR),and ligustrazine preconditioning group(LI).The model of MIRI was produced,Immunohistochemical S-P method was used to detect the expressions of HSP25 and p38MAPK proteins in MIRI tissues.Results:Compared with IR group,the expression of HSP25 protein in LI group increased significantly(P

Background Quality of life(QOL) refers to the person's subjective appraisal of well-being,life satisfaction,health and functional performance. For those stroke patients who cannot finish the scale themselves,we can try the proxy measurement. However,until recently no QOL scale for proxies were induced to Chinese stroke patients. The goal of this research is to translate and test the SIS 3.0 for proxy,and differences between patient and proxy scores. To translate and test the SIS 3.0 for proxy, and analyzed differences between patient and proxy scores.Methods Ten pairs of patients and their proxies were involved in the primary test. Two hundreds and thirty-three pairs were involved in the formal test. We analyzed the validity, responsiveness, reliability and feasibility of the SIS 3.0 for proxy, as well as the validity in proxy assessment. Results The feasibility was sufficient. Both Split reliability and α coefficient were more than 0.8, demonstrating SIS a reliable instrument. SIS had a good content validity with correlation coefficient more than 0.6. Good criterion validity was established by comparing the scores on various domains to standardized measures with P=0.000. Construct validity was also good as indicated by factor analysis. Proxy scores were significantly different across OHS scales which showed domain responsiveness was good. Comparison of patient and proxy responses resulted in no significant difference. Conclusions SIS for proxy is satisfactory for chinese patients. It is feasible to use a proxy respondent to answer questions on the patient's behalf.

Objective:To investigate the clinical characteristics and outcomes in patients with wake-up stroke.Methods:From January 2019 to December 2019, consecutive patients with acute ischemic stroke admitted to the Department of Neurology, Huizhou Municipal Central Hospital were enrolled retrospectively. The modified Rankin Scale was used to evaluate the outcome of patients at 90 d after the onset. 0 to 2 was defined as a good outcome, and >2 was defined as a poor outcome. Multivariate logistic regression analysis was used to determine the factors affecting the poor outcome of patients with wake-up stroke. Results:A total of 356 patients with acute ischemic stroke were enrolled, including 97 (27.2%) wake-up stroke, and 259 (72.8%) non-wake-up stroke. The National Institutes of Health Stroke Scale (NIHSS) score at admission and the proportion of patients with atrial fibrillation, moderate to severe stroke and cardiogenic embolism in the wake-up stroke group were significantly higher than those of the non-wake-up stroke group, and the proportion of patients with small artery occlusion was significantly lower than that in the non-wake-up stroke group (all P<0.05). Moreover, the proportion of patients with poor outcome in the wake-up stroke group was significantly higher than that in the non-wake-up stroke group (36.1% vs. 24.7%; χ2=4.546, P=0.033). In the wake-up stroke group, 62 patients (63.9%) had a good outcome, and 35 (36.1%) had a poor outcome. The NIHSS score at admission and the proportion of patients with atrial fibrillation and moderate to severe stroke in the subgroup with poor outcome were significantly higher than those in the subgroup with good outcome, and the proportion of patients receiving intravenous thrombolysis and mechanical thrombectomy was significantly lower than those in the good outcome subgroup (all P<0.05). Multivariate logistic regression analysis showed that moderate to severe stroke (odds ratio [ OR] 6.674, 95% confidence interval [ CI] 2.223-20.034; P=0.001) was independently associated with the poor outcome in patients with wake-up stroke, while intravenous thrombolysis ( OR 0.102, 95% CI 0.017-0.630; P=0.014) and endovascular mechanical thrombectomy ( OR 0.108, 95% CI 0.023-0.506; P=0.005) were independently associated with the good outcome of patients with wake-up stroke. Conclusions:In patients with wake-up stroke, the proportion of cardioembolism is higher, the clinical symptoms are more serious and the incidence of poor outcome is higher. The severity of stroke at admission is associated with poor short-term outcome, and intravenous thrombolysis and endovascular mechanical thrombectomy can improve the outcome in patients with wake-up stroke.