Production internship is an important teaching tache for undergraduate students to carry out engineering training by using professional skills, and it is a key starting point for fostering application-oriented talents in biotechnology. The Course Group of 'production internship of biotechnology majors' of Binzhou University is investigating application-oriented transformation for local regular colleges and universities, as well as fostering high-level application-oriented talents. By taking green fluorescent protein (GFP) polyclonal antibody as an example, the reform and practice on teaching content, teaching mode, assessment method, continuous improvement of curriculum were carried out. Moreover, the characteristics of the Yellow River Delta-Binzhou Biotechnology & Pharmaceutical Industrial Cluster were taken into account to intensify academic-enterprise cooperation. On one hand, this Course Group designed and rearranged the course contents, carried out essential training through online resources and platforms such as virtual simulation, and recorded, tracked and monitored the progress of production internship through practical testing and software platforms like 'Alumni State'. On the other hand, this Course Group established a practice-and application-oriented assessment method in the process of production internship and a dual evaluation model for continuous improvement. These reform and practices have promoted the training of application-oriented talents in biotechnology, and may serve as a reference for similar courses.
Humans ; Internship and Residency ; Curriculum ; Students ; Biotechnology

Objective To establish an assay method for monohydroxy carbamazepine, the active metabolite of oxcarbazepine, in human plasma. Methods Ornidazole was used as the internal standard. Plasma samples were processed with methanol and analyzed by HPLC. The column was ZORBAX Eclipse XDB-C₁₈(150 mm×4.6 mm, 5 μm) with the mobile phase of water-acetonitrile (80∶20, V/V) at a flow rate of 1.0 ml/min. Dual wavelength detection is applied. The detection wavelength of monohydroxy carbamazepine was set at 192 nm and ornidazole at 318 nm. Results There was an excellent liner relationship for monohydroxy carbamazepine from 2 to 50 μg/ml(r= 0.998 6). The limit of quantification was 2 μg/ml with the range of accuracy between 95.57% and 100.59%. The RSD of intra-day and inter-day precisions were less than 15%. The average extraction recovery rate of MHC and internal standard were in the range of 89.62% to 98.76%. The RSD of stability was less than 6%. Conclusion This method is specific, sensitive, and easy to operate. It is suitable for the clinical assay of monohydroxy carbamazepine in human plasma.

Objective:To understand the present situation of self-acceptance of elderly in nursing home and explore its influencing factors, so as to provide basis for improving self-acceptance level of elderly in nursing home.Methods:Totally 415 elderly people in 3 Medical and nursing homes in Fuyang city of Anhui province were selected by Self-Acceptance Scale, and influencing factors were analyzed.Results:The total score of Self-Acceptance Scale of the elderly in nursing home was (44.11±7.25) points, the score rate was 67.86% (44.11/65), in the middle level; the results of multiple linear stepwise regression showed that the relationship with children, hobbies, number of children, length of stay in nursing home, frequency of visits and age were the main influencing factors ( F value was 48.228, P<0.001). Conclusion:The self-acceptance of the elderly in nursing home is in the middle level. The nursing home or social should pay attention to the higher age group and the newly admitted elderly in nursing home, give such people timely psychological guidance, help them to plan their later life, cultivate their interests and hobbies through recreational activities, and take corresponding measures to harmonize their relationship with their children in order to improve their self-acceptance level and promote mental health.

Objective Based on the data of gemcitabine adverse reactions (ADR), the characteristics and patterns of ADR occurrence were analyzed. The guidance for the rational clinical use of gemcitabine was provided. Methods From our information system, the patients treated by gemcitabine chemotherapy from September 2008 to September 2018 were selected and their related ADRs were summarized and statistically analyzed by using SPSS 18.0. Results Among the 750 cases, there were 312 ADRs related to gemcitabine treatment, of which the incidence of ADR was higher in female patients than in male patients (48.68% vs 36.77%) and the highest incidence in the age group of 50-69 years (44.14%). The patients with lower status (KPS) scores were more sensitive to chemotherapeutic drugs, and more likely to develop ADR. During the combination therapy, gemcitabine+paclitaxel chemotherapy had the highest incidence of ADR (61.54%) and the highest incidence of ADR in thymic carcinoma (62.50%), followed by hematological and reproductive system tumors (58.62% and 57.14%, respectively); ADR involvement in organs/systems is mainly caused by hematological toxicity. The bone marrow suppression is common, followed by digestive system damage. The major clinical symptoms were nausea and vomit. Conclusion Gemcitabine-related ADR has a large individualized difference and is affected by many factors. Chemotherapy should be used according to individual conditions to improve clinical safety and rational use of drugs.

To analyze the clinical, molecular genetic and biochemical characteristics of a family diagnosed with creatine deficiency syndrome (CDS) due to SLC6A8 gene mutation diagnosed in Department of Pediatrics, the First Affiliated Hospital of Xinxiang Medical University in September 2018.The boy was referred at the age of 13 for intermittent vomiting and symptoms were relieved after symptomatic treatment, 7 months later, the patient was diagnosed again.He ate less and constipated as an infant, and had intermittent vomiting and severe constipation from 12 years old.He could walk at the age of 1 year and 6 months, could speak his first word at 2 years old, and only spoke simple sentences at the diagnosis.He rarely communicated with his parents.His brother has severe intellectual deficiency and seizures.His mother has cognitive impairment and had repeated stillbirths and miscarriages.When visiting the doctor, his weight was 22 kg (< P3), height was 131 cm (< P3), and head circumference was 52 cm.Besides, he had severe mental retardation and sluggish expression, poor communication, broad forehead, musculoskeletal atrophy and arched foots.His brain magnetic resonance imaging showed mild enlargement of the lateral ventricles.Magnetic resonance spectroscopy showed normal signals in the bilateral hippocampuses.The boy′s exon sequencing related to intellectual deficiency revealed the hemizygous mutation of SLC6A8 gene c. 626(exon3)-c.627(exon3)delCT.His mother and brother were carriers of the variant gene, but his father and grandmother had no such mutation.The urinary creatine to creatinine ratio was elevated but guanidinoacetate was normal, it was further confirmed that CDS can be caused by the novel mutation c. 626(exon3)-c.627(exon3) delCT in SLC6A8.In clinical practice, boy with mental retardation of unknown origin and often have gastrointestinal symptoms, especially when feeding difficulties and growth retardation occur in infancy, the early diagnosis of CDS can be achieved by examination of urinary creatine to creatinine ratio and gene detection.

Objective To establish an assay method for unbound teicoplanin in plasma by centrifugal ultrafiltration combined with ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Methods Protein was removed from plasma by a Centrifree^® ultrafiltration device. The ultrafiltrate was injected to determine the unbound concentration of teicoplanin. Endeadvorsil^TM C₁₈ column (1.8 μm, 50 mm×2.1 mm) was used with gradient elution of acetonitrile and 0.02 mol/L ammonium acetate solution (containing 0.1% formic acid). The detection was performed on a triple-quadrupole tandem mass spectrometer by multiple reaction monitoring (MRM)mode via electro spray ionization (ESI). Results The calibration curve of unbound teicoplanin in plasma was linear over the range of 0.10 to 8.00 μg/ml (r=0.999). The intra-assay precision and the inter-assay precision of samples didn't exceed 7.00%. The average relative recovery ratio was 97.9%, and the matrix effect factor was 0.97. The samples had good stability after being stored at room temperature for 10 h or at −20 ℃ for 15 days, and freeze-thawed 3 times (RSDs were all within 6.50%). Conclusion This method is convenient, fast, sensitive and accurate. It provided a basis for clinical development of teicoplanin unbound concentration monitoring.

Objective To explore the clinical effect of training assisted by a lower limb rehabilitation robot on the bladder and intestinal function of paraplegic spinal cord injury survivors. Methods Thirty-eight paraplegic patients with spinal cord injury were divided according to their admission order into an experimental group ( n=19) and a control group (n=19). Both groups were given conventional rehabilitation training, while the experimental group was additionally provided with robot-assisted lower limb training in three stages:adaptation, training and con-solidation. It lasted 30 minutes daily, 5 days per week for 12 weeks. Before and after the training, an urodynamics examination system was used to evaluate the maximum urine flow, bladder capacity, residual urine volume, bladder pressure and detrusor pressure. Colon transit time, mean rectal pressure and intestinal function were measured using the colon transit test, a mean rectal pressure test, and the Functional Independence Measure ( FIM) scale respective-ly. Results The average bladder volume, maximum urine flow rate, average urine flow rate, detrusor pressure, bladder compliance, average rectal pressure and intestinal FIM score of the robot training group after training were all significantly better than before the training, as were the average residual urine volume and colon transit time. After the training, the average bladder volume, maximum urine flow rate, average urine flow rate, detrusor pressure, bladder compliance and average rectal pressure of the robot training group were all significantly higher than those of the control group, while the average residual urine volume and colon transit time were significantly smaller. Then, 32％ of the patients in the experimental group achieved no less than 6 points for their average FIM score, significantly higher than in the control group. Conclusion Robot-assisted lower limb training combined with comprehensive rehabilitation training can effectively improve the bladder and intestinal function of paraplegic patients after a spinal cord injury.

Objective To study the expression of visfatin and related mRNAs in myocardium of acute myocardial ischemia in rats,and to explore the role of both in acute myocardial ischemia model and to propose new ideas for clinical cardiovascular diseases.Methods Wistar rats (n=20) were randomly divided into sham operation control group and acute myocardial ischemia groups.Left anterior descending (LAD) of coronary artery was ligated to establish the model of acute myocardial ischemia.The total myocardium proteins were collected 12 h after operation.Western blot was used to observe the expression changes of visfatin in myocardium.The expressions of miR-124 and miR-327 in myocardium were detected by real-time PCR.Results The levels of serum CK and LDH in the model group of myocardial ischemia rats were elevated [(46.20±7.12) vs (72.19±9.38)kU/L;(2542.23±173.54) vs (6428.76±237.81)U/L,P＜0.05)].The visfatin expression levels in the rats of acute myocardial ischemia groups were higher than those in controls [(0.804±0.042) vs (1.046±0.031),P＜0.05].MiR-124 was up regulated and miR-327 was down regulated in acute ischemia myocardium.Cndusion The up-regulation of miR-124,the down-regulation of miR-327 and the increase of visfatin expression may be related to acute myocardial ischemia.The changes of visfatin expression may be regulated by miR-124 and miR-327 posttranscriptional levels.

Objective To evaluate clinical value of interventional radiology in difficult extraction of stuck tunnel hemodialysis catheter (TCC).Methods Maintenance hemodialysis patients who required removal of stuck TCC in our hospital from June 2015 to June 2017 were enrolled.Medical history and preoperative examinations were carefully evaluated.Patients were pulled rotationally the catheter under the fluoroscopy guidance and exchanged the TCC.The information and procedure-related complication were recorded.Balloon dilation or snare technique would be used when failed to remove the stuck catheter.Results A total of 22 patients were enrolled,with an average age of (67.2±18.5) years,a median dialysis time of 54(18,106) months,and catheter indwelling time of 37(14,82) months.All patients were successfully removed the stuck catheter,among which 18 cases (81.8％) had fibrin sheath and 13 cases with sheath calcification.TCC mainly stuck in the junction of internal jugular vein and innominate vein,middle and lower innominate vein and middle part of superior vena cava.No procedure-related major complication occurred.Sixteen cases (72.7％) complained of transient substernal extraction pain and 1 case occurred groin hematoma during operation.Conclusions Catheter incarceration problem should be alert to the hemodialysis patients with catheter indwelling time of more than 2-3 years.The minimally invasive interventional techniques including rotationally pull-out under fluoroscopy guidance and balloon angioplasty are safe and effective in removal of the stuck TCC.

Objective To establish method of constructing lentiviral vectors to express microRNA (miRNA) ''tough decoy''(TuD)and to detect the effects of the TuD on cellular endogenous miRNA level and cellular phenotypes. Methods Two-step cloning strategy was utilized to first generate a universal miRNA TuD frame vector,followed by con-structing the TuD expression vector specially targeting miR-203. The package of the recombinant lentivirus was per-formed in 293T cells. Then the rat bone marrow mesenchymal stem cells(BM-MSCs)were infected by the miR-203 TuD expression lentivirus. The pSIH1-H1-copGFP vector was also packaged and the BM-MSCs infected by this lentivirus were served as control. Endogenous miR-203 level in BM-MSCs was measured by quantitative RT-PCR,and cellular vi-ability and apoptosis were detected by CCK-8 test and Annexin V-PI staining respectively. Results The miR-203 TuD expression vector was successfully constructed and inserted sequence was validated. At the 3rd,6th and 9th days after in-fected by the miR-203 TuD expression lentivirus,rat BM-MSCs exhibited a repressed endogenous miR-203 level. The miR-203 TuD also promoted viability and inhibited apoptosis of BM-MSCs. All these differences between miR-203 TuD group and control group were statistically significant. Conclusion The two-step cloning method for the construction of miRNA TuD expression vector is simple and efficient. The miRNA TuD can effectively suppress the level of the target miRNA and affect cellular phenotypes.