According to relevant national laws and regulations, practitioner training was included into laboratory animal science teaching reform.By adjusting the training content and teaching method and use of animal models of typical human diseases, the transformation of training mode was realized and improved.By the assessment of basic theory in combination with practical operation, the thinking ability and hands-on skill of the practitioners are much improved. Through classroom instruction, experimental teaching, quality assessment and tracking survey, the evaluating process of the training quality of training teaching is performed.Therefore, the teaching reform of the laboratory animal science based on the training of practitioners is established.

ObjectiveUsing large sample clinical study to reevaluate the safety and efficacy of recombinant human growth hormone(ANSOMONE) in treating pediatric growth hormone deficiency(GHD).Methods1012 cases were administered with ANSOMONE at a dose of 0.1 IU · kg-1 · d -1 in bedtime for 6 months.The alteration of mean height,growth rate,height standard deviation scores,bone age,and anti-GH antibody and occurrence of adverse effects were observed.ResultsThe growth rate was promoted from ( 2.50 ± 0.85 ) cm/year to ( 13.44 ± 4.27 ) cm/year,the net growth rate was( 10.94 ±3.69)cm/year.There was significant difference between the pre-treatment and post-treatment patients ( P ＜ 0.01 ).The post-treatment anti-GH antibody positive rate was 13.34％,which had no obvious effect on the efficacy.Few patients exert transient inflammation reactions,which disappeared with any treatment.ConclusionANSOMONE had significant efficacy,and could promote the height increase obviously.It had good safety without any obvious side effect and early-maturing effect.

Objective To evaluate the effect of near infrared heptamethine cyanine dye IR-783-mediated specific tumor imaging in spontaneous tumor of dogs .Methods IR-783 was intraperitoneally injected to nude mice models of transplanted tumor in a dose of 5μmol/kg.The metabolic time course of IR-783 was detected by in vivo imager .Based on the results of above observation , IR-783 was injected to dogs with spontaneous tumor in a dose of 1.5μmol/kg.The site of intravenous injection was the hind leg .Tumor and peri-tumoral tissues were removed at 5 days after IR-783-injection for fluorescence imaging , pathology and frozen section fluorescence examinations .Results After i.p.IR-783 injection to nude mice models of transplanted tumor , the transplanted tumor tissues of nude mice had stronger specific fluorescence than normal tissues by imaging at 8 days after injection .After i.v.IR-783 injection to four dogs with spontaneous tumor , the fluorescence signal in the tumor tissues was stronger than that in the normal tissues at 5 days after injection .Conclusions Near infrared fluorescent dye IR-783 could be specifically taken up by tumor tissues , and can be used for specific diagnosis of tumor.It has an important clinical application prospect .

Upper urinary tract calculi with infection is a quite difficult acute urologic emergency.And what is more, upper urinary tract obstruction after radical cystectomy following urinary diversion may be fatal in the vulnerable patients with this kind of situation.Postoperative anatomy and other factors affect the upper urinary tract calculi, and urinary tract infection greatly increased the risk.But it is particularly difficult to handle with patients with poor general condition and septic shock treatment, so how to optimize the selection program is worth further studying.A 65-year-old man was admitted to hospital for combining with renal and ureteral calculi after radical cystectomy (Bricker) in March 2017.The patient underwent transurethral resection of bladder tumor in the outer court because of the discovery of bladder tumor one year before, The postoperative pathology was high-grade papillary urothelial carcinoma and he underwent regular irrigation of the bladder.Because of tumor recurrence, he came to Peking University Third Hospital for further treatment in August 2016, and the examination of urinary tract CT indicated bladder can-cer;for bilateral multiple renal pelvis and renal calices calculi, he was given laparoscopic radical cystecto-my.After four months, due to sudden chill fever, he was hospitalized.After definite diagnosis, anti-inflammatory treatment combined with left nephrostomy was given.The two-stage lithotripsy was performed.After expansion of the original left renal fistula to 24 F(1 F=0.33 mm), 24 F sheath was inducted into the kidney.We introduced a flexible ureteroscopy into the channel down to the ureteral obstruction, where we found a 1 cm oval black brown stone obstructed.A guide wire was inducted through the stones down to ileal conduit in the right lower quadrant.Through the ureteroscopy, we made ureteral calculi broken into powder, and replaced the residual stones into the renal pelvis.Then we used the nephroscopy for lithotropsy in the pelvis, and finally we introduced a flexible cystoscopy for the residual stone.The operation was successful, the operation time was 181 min, and intraoperative bleeding was 10 mL.After operation, no residual stones were found in kidney, ureter, bladder (KUB) plain films.No operation complications were related to the urinary tract.Therefore, multiple endoscopy with combination therapy of upper urinary tract calculi following urinary diversion is safe and effective, and the reasonable choice with the advantages of each instrument can improve the stone clearance rate and shorten the operation time.This can be used as a useful complement to traditional treatment.

Chemotherapy, Adjuvant ; Gastrectomy ; Humans ; Microsatellite Repeats ; Neoadjuvant Therapy ; Stomach Neoplasms/pathology*

Altitude ; Heart Rate ; Humans ; Oxygen ; Oxygen Consumption

Objective To construct miRNA-29b1 gene knockout mice based on CRISPR/Cas9 technology. Methods To design and synthesize sgRNA according to the miRNA-29b1 sequence in Genbank .sgRNA and Cas9 were transcribed to RNA in vitro, these RNA were then microinjected into zygotes of C 57BL/6 mice.After mouse birth, the genome DNA was extracted and sequenced to identify its genotype; meanwhile , real-time PCR was used to assay the expression of miRNA-29b1 in the heart, liver, spleen, lung and kidney of mutated mice .Result A 20 bp sgRNA targeted on miRNA-29b1 was synthesized and transcribed to RNA with Cas 9.After microinjection, miRNA-29b1 gene-mutated mice were obtained.The sequencing results showed that there were two types of genotype for the mutated mice , one was 10 bp deletion, and another was 23 bp deletion accompanied with a 3 bp insertion.Compared with the wild-type mice, the expression of miRNA-29b1 in the heart, liver, spleen, lung and kidney was reduced significantly .Conclusions miRNA-29b1 gene knockout mice are constructed successfully by using CRISPR /Cas9 technology.

Objective To knockout Rag2 and IL2rg genes and construct severe combined immunodeficiency mice based on CRISPR/Cas9 technology. Method Design and synthesis of 25 bp sgRNA were made according to the Rag2 and IL2rg sequences in Genbank. After annealing, sgRNA was cloned into pX330 vector. Recombination plasmid Rag2?sgRNA, IL2rg?sgRN and Cas9 were then transcribed into RNA, these RNA were microinjected into zygotes and the zygotes were transplanted into recipient ICR mice. F0 founders were born and mutated F0 founders mated with wild type mice to obtain F1 generation heterozygous mice. Mutated F1 mice were crossed and got F2 generation homozygous mice. Genotype and phenotype of the knockout mice were identified by sequencing, flow cytometry and xenograft model. Results Rag2?sgRNA and IL2rg?sgRNA recombination plasmids were constructed and transcribed into RNA. After microinjection and mat? ing, F0 founders were born and F2 homozygous mice were obtained. The results of sequencing showed that there were two types of genotype in IL2rg gene, 10 bp or 11 bp deletion;however, there was only one genotype in Rag2 gene, which was 8 bp deletion. Compared with wild?type BALB/c mice, the number of CD3 +, B220 + and NKp46 + cells in peripheral blood of the knockout mice was reduced significantly. After inoculation of human breast cancer cell line SKBR?2HL cells, tumor size in the xenograft mouse model was increased gradually along with time extension. Conclusion CRISPR/Cas9 is an efficient way to mutate Rag2 and IL2rg gene in mice in vivo, leading to aberrant T cells, B cells and NK cells.

Adolescent ; Altitude ; Brain ; drug effects ; Codonopsis ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Tablets

OBJECTIVETo investigate the effects of simple hypobaric hypoxia on parameters of hematology and blood rheology in order to establish a rat model of simulated high altitude polycythemia (HAPC) for the study of pathophysiologic mechanisms and medical prevention and treatment of HAPC.

METHODSForty-eight male Wistar rats were randomly divided into three normal control groups and three hypoxia model groups. Normal control group rats were bred in normoxia conditions, and hypoxia group rats were subjected to hypoxic exposure for 8 hours per day at simulated 5 500 m high altitude in a hypobaric chamber. After hypoxic exposure for 2, 4, 12 weeks, one group of normal control and hypoxia model rats were killed and blood was collected, respectively. Then parameters of erythrocyte and blood rheology were examined.

RESULTSMucous membrane of hypoxia model rats showed obviously cyanosis after 2 weeks hypoxic exposure. Hemoglobin concentration of hypoxia model rats were beyond 210 g/L after 2 weeks, 4 weeks and 12 weeks hypoxia exposure and significantly increased than that of normal control rats respectively. Besides, RBC counts, hematocrit, whole blood viscosity, erythrocyte aggregation index of hypoxia model rats were all notably higher than those of normal control rats respectively.

CONCLUSIONA rat model of high altitude polycythemia can be rapidly established by hypobaric hypoxia exposure at simulated 5 500 m high altitude for 8 hours daily.

Altitude ; Altitude Sickness ; Animals ; Disease Models, Animal ; Erythrocyte Count ; Hematocrit ; Hypoxia ; Male ; Polycythemia ; pathology ; Rats ; Rats, Wistar