OBJECTIVE:To investigate the preparing process and method of quality control of the podophyllotoxin film former METHODS:To sift the formula of podophyllotoxin film former with orthogonal function and to determine the content by second derivative UV-spectrophotometry RESULTS:The peak area of podophyllotoxin against the concentration was linear with a correlation coefficient of 0 9 999 over the range of 80～180?g/ml and detecting at wavelength 301 5nm～315nm CONC_LUSION:The preparing process of podophyllotoxin film former is simple and the method of quality control is feasible

BACKGROUND:Because of convenient source, multi-lineage differentiation and low immunogenicity, bone marrow mesenchymal stem cels are the ideal cel type to serve as vectors of transgenic cels in pain management. However, the replicative senescence and smal amount of cels obtained from the bone marrow restrict the application of bone marrow mesenchymal stem cels in pain research. OBJECTIVE:To construct human telomerase reverse transcriptase (hTERT)-immortalized rat bone marrow mesenchymal stem cels as transgenic celular vectors for pain therapy. METHODS:Bone marrow mesenchymal stem cels were obtained from whole rat bone marrow, and then transfected with a lentivirus containing the hTERT (pLV-Puro-EF1α-hTERT) folowed by puromycin selection. hTERT expression and telomerase activity in these transfected cels were determined by RT-PCR and TRAP. Morphological changes, capacity of cel growth and multi-lineage differentiation, chromosome karyotype and tumorigenicity were observed in vitro. Moreover, the expression of cel surface molecule, Nestin, MHC-I and MHC-II in transfected cels were also detected by flow cytometry and immunocytochemistry. RESULTS AND CONCLUSION: The bone marrow mesenchymal stem cels geneticaly modified by hTERT could be cultured and passaged through 30 generations in vitro. Compared to the primary and negative transfected cels, the hTERT-modified bone marrow mesenchymal stem cels showed higher expression of hTERT mRNA, telomerase activity and cel proliferation. Most of transfected cels stayed at G2/M and S stages. The proliferation index of the transfected cels were increased dramaticaly. The positive rates of CD29, CD44 and CD90 were over 70%, but the positive rates of CD34 and CD45 were less than 5%. Transfected cels were positive for Nestin in the cytoplasm, but negative for MHC-1 and MHC-11. In addition, this cel line continued to exhibit the characteristics of fibroblastic bone marrow mesenchymal stem cels, including phenotype, differentiation into osteoblasts, adipocytes and neuron-like cels. No chromosome abnormality and tumor formation were observed in this experiment. Taken together, these data suggests that the rat bone marrow mesenchymal stem cels immortalized by hTERT gene are constructed successfuly and stil maintain major stem cels characteristics, which provide safe and stable cel vectors as research base for pain therapy.

Objective To study the effects of classification-partition-distribution emergency nursing management for severe trauma patients. Method A total of 60 patients from June 2014 to May 2015 were set as control group receiving common nursing and other 62 patients from June 2015 to June 2016 as observation group treated with emergency hierarchical partition and triage nursing. Result The treatment success rate in the observation group were both significantly higher than that of the control group (P<0.05). Conclusion Classification-partition-distribution emergency nursing management for severe trauma patients can increase treatment success rate .

Objective To extract ?-glucosidase inhibitors from Potentilla anserina L. Methods Water-decocting method and water-decocting ethanol-precipitation were applied to obtain water extract,sodden ointment and polysaccharides of Potentilla anserina L. The colorimetry of pNPG was used for the determination of the activity of alpha-glucosidase pure enzyme and alpha-glucosidase from isolated small intestines of natural ICR mice. Results The extracts of Potentilla anserina L. had obvious inhibitory effect on the pure enzyme of ?-glucosidase. The water extract and sodden ointment also had inhibitory effect on ?-glucosidase from isolated small intestine in natural ICR mice,the effect being in a dose-dependant manner. Conclusion The extracts of Potentilla anserina L. have the inhibitor-like effect on ?-glucosidase,and the active position is probably in the sodden ointment.

Objective To evaluate the value of detecting HCMV-DNA in urine in the diseases of newborn babies ,by comparing the result between HCMV-DNA in urine and HCMV-IgM antibody in blood in different diseases of the newborn babies .Methods The urine and blood samples were collected from 1 520 infants who were in hospital of the neonate department from January 2013 to December 2014 .The HCMV-DNA in urine was examined by fluorescence quantitative polymerase chain reaction(FQ-PCR) .And HCMV-IgM antibody in blood was examined by the method of chemiluminescence(ECL) .Results In the 1 520 cases ,153 had de-tectable HCMV-DNA in their urine samples with a positive rate 10 .07% ,while only 4 cases were positive of the HCMV-IgM anti-body in blood with a positive rate 0 .27% .The two groups was statistically significant difference (P<0 .05) .In the infant diseases of 1 520 cases ,the positive rate of HCMV-DNA in urine was 82 .6% of the hepatitis syndrome while the positive rate of HCMV-IgM antibody in blood was 3 .85% .The positive rate of HCMV-DNA in urine was 8 .95% in the jaundice symptoms ,while the posi-tive rate HCMV-DNA in urine was 4 .62% in the pulmonary infection .Each group of disease was statistically significant difference in the positive rate of HCMV-DNA (P<0 .05) .Conclusion The detection of HCMV-DNA in urine is more beneficial to the diag-nosis of HCM V infection of infants than the detection of HCM V-IgM antibody in blood ,especially in hepatitis syndrome ,neonatal jaundice and pulmonary infection .

Objective To investigate the effect of periurethral injection of insulin-like growth factor(IGF)-Ⅰ on the expression of IGF-Ⅰ and IGF-Ⅱ mRNA during regeneration period following urethral sphincter muscle injury in female rats. Methods Model of urethral sphincter muscle injury was made in female virgin SD rats (n=50) by intravaginal balloon inflation. Then the rats were divid-ed randomly into treatment group (n= 25) and control group (n= 25), treatment group accepted peri-urethral injection of 1.0μg human IGF-Ⅱ to the middle urethral muscle, control group accepted nor-mal saline injection. Five rats in each group were sacrificed at 2, 4, 6, 8, 14 day respectively and the whole urethra specimens were processed for RT-PCR to detect the expression of IGF-Ⅰ ,Ⅱ mRNA. A normal control group (n = 5) was set without intravaginal balloon inflation and injection. Results The expression of IGF-Ⅰ mRNA in control group increased at day 4, 6, 8, 14, the IGF-Ⅰ/β-actin ra-tios were 0. 58±0.15, 1.73±0.31, 2.30±0.29, 0. 46±0. 06. The expression of IGF-Ⅰ mRNA in treatment group increased at all time points, as 0. 69±0.21, 1.45±0.17, 2.25±0.45, 2.90±0.49, 1.92±0. 31. The difference was significant on day 4, 14(P<0.01), and day 8 (P<0.05) compared with the control group. The expression of IGF-Ⅱ mRNA in control group increased at day 4, 6, 8, as 0.42±0. 14, 1.51±0. 59, 1.31±1.04. The expression of IGF-Ⅱ mRNA in treatment group in-creased at day 4, 6, 8, 14, as 1.04±0.23, 1.94±0.29, 1.75±0.41, 0. 81±0.15. The significant difference was noted on day 4 (P<0. 01)compared with the control group. No expression of IGF-Ⅰand Ⅱ mRNA in the normal control group. Conclusions The expression of endogenous IGF-Ⅰ and IGF-Ⅱ mRNA was up-regulated by periurethral injection of IGF-Ⅰ during regeneration period follow-ing urethral sphincter muscle injury in female rat. Our findings suggest that IGF-Ⅰ facilitates the re-generation of the urethral muscles and may play a role in treatment of stress urinary incontinence in-duced by urethral sphincter muscle dysfunction.

Objective To investigate the P2X3 receptor expression in L6-S1 dorsal root ganglion (DRG)and bladder detrusor in a rat model of neurogenic bladder and urethra. Methods Eighty Sprague-Dawley rats wererecruited and randomly divided into a sacral injury group, a suprasacral injury group and a control group. Spinal tran-section was performed to establish the animal model of neurogenic bladder and urethra in rats of the sacral injurygroup and suprasacral injury group. Check the P2X3 receptor expression in DBG and bladder detrusor among thethree groups by Western blot test at 20 days after model establishment. Results P2X3 receptor expression in L6-S1DRG of sacral injury group was significantly less than that of the suprasacral injury group, which was in turn signifi-cantly higher than that of the control group. P2X3 receptor expression in bladder detrusor of sacral injury group wassignificantly lower than that of the suprasacral injury group, which was in turn significantly higher than that of thecontrol group. Conclusion There was close relationship between P2X3 receptor expression and dysfunction of blad-der and urethra.

AIM:To investigate the expression and significance of thrombospondin-1(TSP-1)in left ventricular myocardium of type 2 diabetic cardiomyopathy(DCM).METHODS:The rat model of DCM was established by eating a high-fat diet together with injection of low dose streptozocin(30 mg/kg)intrapertoneally.After 12 weeks,the content of collagen was quantified by Masson staining.The mRNA level of TSP-1 was determined by quantification real-time RT-PCR,while the protein level of TSP-1 was analyzed by Western blotting and immunohistochemistry.RESULTS:Compared with the control group,the content of collagen in the DCM group was increased greatly(11.01?3.05 vs 16.92?3.18,P

Objective To investigate the perinatal outcomes after elective neurosurgery in pregnant women complicated by space-occupying lesions in saddle area.Methods Clinical data were retrospectively collected and analyzed from five gravidas with space-occupying lesions in saddle area,who underwent neurosurgery at Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine from June 2017 to February 2018,including gestational age,perioperative management,surgical procedures,postoperative complications,pregnancy outcomes and pathological results.Results (1) Progressively deteriorated impaired vision was the chief complaint of all five patients.Space-occupying lesions in saddle area were detected by cranial MRI (plain scan).The surgical indication was clear as there was compression on optic nerve and a risk of blindness due to optic nerve atrophy,Two patients were operated in the third trimester and three in the second trimester.Postoperative pathology showed there were two cases with meningioma,two with pituitary macro adenoma and one with abscess of pituitary.Only one patient had mild postoperative cerebrospinal fluid rhinorrhea,which was improved three months later,and no complications were reported in the other four cases.All five patients had obvious improvement of vision after surgical intervention and no relapse was reported during a follow-up of 4-12 months.(2) Four gravidas continued their pregnancies to full or nearly full term,among which three underwent cesarean section due to obstetrical indications (one breech pregnancy,one twin pregnancy with scarred uterus and one complicated by hypertensive diseases of pregnancy with poor blood pressure control) and one delivered vaginally.Postpartum involution of the uterus was well in all of the four cases 42 d after delivery.The women with pituitary abscess terminated the pregnancy due to intraamniotic injection of ethacridine lactate at 20 weeks of gestation.Five neonates were all born with Apgar scores of 10 at 1,5 and 10 min without any visible defects at birth and all grew and developed normally at 4-12 months after birth.Conclusions For gravidas with space-occupying lesions in saddle area,it is necessary to weigh the pros and cons and choose an individualized treatment plan.Resection of saddle area lesions in the second or third trimester backed by a multidisciplinary team is a safe and feasible procedure if postponing the operation until delivery may cause irreversible or even life-threatening consequences.

Objective To evaluate the effect of adenosine receptor antagonist SCH442416 on the expression of Kir2.1,Kir4.1 and TASK-1 in rat Müller cell at an elevated hydrostatic pressure in vitro.Methods Thirty SPF Sprague Dawley rats were purchased from Shanghai Slack Laboratory Animals Ltd.Cultured Müller cells were divided into normal control group (n =6),40 mmHg/24 hours (1 mmHg =0.133 kPa) group (n =6) and adenosine + SCH442416 intervention group (n =6).Müller cells were treated with 40 mmHg pressure for 24 hours in 40 mmHg/24 hours group,and Müller cells were treated with 40 mmHg pressure for 24 hours + 10 μ mol/L adenosine + 100 nmol/L SCH442416 in adenosine + SCH442416 intervention group.The real-time PCR,Western blot,whole-cell patch-clamp recordings and immunohistochemistry were used to detect Kir2.1,Kir4.1 and TASK-1 expression and Müller cells Kir currents.The experimental procedures were in accordance with the National Institutes of Health (NIH) guidelines for the Care and Use of Laboratory,and follow the 3R principle.Results Western blot assay showed that,following 40 mmHg pressure cultured for 24 hours,the expression of Kir4.1 and TASK-1 protein in Müller cell were significantly decreased by 38.6％ and 52.6％ compared with the normal control group,with significant differences between the two groups (both at P =0.000);Kir2.1 protein expression decreased by 14.7％,with insignificant difference between the two groups (P =0.082).Kir4.1 and TASK protein expression in adenosine + SCH442416 intervention group was increased by 60.7％ and 61.4％ compared with the 40 mmHg/24 hours group,with significant differences between the two groups (both at P =0.000);Kir2.1 protein expression in adenosine + SCH442416 intervention group was increased by 8.8％ compared with the 40 mmHg/24 hours group,with insignificant difference between them (P =0.354).Real-time PCR assay showed that,following 40 mmHg pressure cultured for 24 hours,Kir2.1,Kir4.1 and TASK-1 mRNA expression in Müller cells were significantly decreased compared with the normal control group,with significant differences between the two groups (P =0.047,0.001,0.000);Kir4.1 and TASK-1 mRNA expression in Müller cells in the adenosine + SCH442416 intervention group was significantly increased compared with the 40 mmHg/24 hours group,with significant differences between the two groups (P =0.038,0.030);however,there is no significant change in Kir2.1 mRNA expression (P =0.612).Conclusions SCH442416 upregulates the expression of Kir4.1 and TASK-1 mRNA and protein,but weakly affects the expression of Kir2.1.