Acute alveolar injury was induced in rats by subcutaneous injection of Nnitroso-N-methylurethane (NMU) 6mg/kg body wt. NMU decreased the content of surfactent phosphoLipids, and increased the protein of alveolar lavage return of rats after 2 days of the injection. Such changes were not observed in lamellar bodies and microsomcs. Composition of phospholipids also changed in all lung fractions it showed that the ratio of PG/PI was decreased significantly in response to NMU. The molecular species profiles of both acidic phospholipids in the lung fractions were distinctly different from each other and the main molecular species were attered during the acute alveolar injury.

BACKGROUND:For treatment of central nervous system diseases, neural stem cel s (NSCs) or bone marrow stromal stem cel s (BMSCs) can be transplanted into the brain, but there are less reports to compare the effects of two kinds of stem cel transplantation. OBJECTIVE:To explore the effect of midbrain NSCs and BMSCs on the behavior and brain morphology of rats with Parkinson’s disease. METHODS:Fifty-eight Sprague-Dawley rats were enrol ed to establish Parkinson’s disease models, and then randomly divided into three groups, which were treated with 5μL midbrain NSCs (n=20), 5μL BMSCs (n=20) and 5μL normal saline (n=18) via two coordinate points of the right striatum at 3 weeks after modeling, respectively. At 5 months after transplantation, the rats underwent intraperitoneal injection of apomorphine to observe behavioral changes, and then, the striatum was taken for immunohistochemistry staining. RESULTS AND CONCLUSION:The number of rotations was reduced significantly in the BMSCs and midbrain NSCs groups at 5 months after transplantation (P<0.05), which was significantly lower than that in the normal saline group (P<0.05). However, there was no significant difference between the BMSCs and NSCs groups (P>0.05). In the BMSCs group, BrdU/Nestin positive cel s were seen in the brain stratium at 1 week after transplantation;BrdU/GFAP and BrdU/NSE positive cel s as wel as TH positive cel s rather than BrdU/TH positive cel s were found in the brain stratium at 1 month after transplantation;after that, the number of BrdU/Nestin positive cel s was reduced gradual y and disappeared ultimately, but there were stil a certain number of BrdU/GFAP and BrdU/NSE positive cel s, especial y the former ones. Meanwhile, the NSCs group also had a similar situation, but no double-labeled cel s were in the normal saline group. These findings indicate that midbrain NSCs and BMSCs transplantation can both improve the behavior of Parkinson’s disease rats, and differentiate into neurons, astrocytes and dopaminergic neurons.

OBJECTIVE: To investigate the use of antibiotics in our hospital. METHODS: All the prescriptions in Jan., Apr., July and Oct. in 2006 in dispensary for western medicine were reviewed, from which, 6 605 (17.26%) antibiotic prescriptions were selected. The categories and kinds of antibiotics were investigated; meanwhile the administration and dosage, course of treatment and drug combination were analyzed based on diagnosis and patient's age. RESULTS: The antibiotics used in 6 605 prescriptions totaled 7 categories. 87.80% used antibacterial drugs accorded with diagnose; 56.46% of the antibacterial drugs were administered orally; in terms of dosage and administration, more than 95% were up to the standard; the course of treatment in 86.99% ranged from 3 to 7 days; 91.04% of the antibacterial drugs used in single kind, about 8.80% in two kinds, and only 0.17% in three kinds concomitantly. CONCLUSION: The use of antibacterial drugs in our hospital is rational on the whole, yet it is far from perfect. Measures should be taken to further improve the rational drug use level.

AIM: To obtain the glycopohorin A (GPA) cDNA and construct the target gene in yeast two-hybrid.METHODS: About 410 bp cDNA fragment was amplified from K562 cell by RT-PCR.After being sequenced, the GPA gene fragment was cloned into EcoR -Ⅰ- Pst Ⅰ site of pbridge to form BD ends in yeast two-hybrid system. The recombinant plasmid was transfered into yeast AH109, and the expression in the yeast was also examined. RESULTS: The amino acid sequence encoded by cloned cDNA was mostly the same as reported GPA, and about 1 mm white yeast clone grew in the selective medium after 3 d.CONCLUSION: pbridge-GPA has nontoxic to the yeast, which can serve as a target gene in yeast two-hybrid system.

Objective To compare the effects of telmisartan and (or) amlodipine on the reversal left ventricular re-modeling in two-kidney one clip hypertensive rats. Methods A total of 50 healthy male SD rats were randomly divided into 5 groups (n=10):two-kidney one clip renal hypertensive (2KIC) model group, sham group, telmisartan (10 mg/kg) group, am-lodipine (2.5 mg/kg) group and telmisartan (10 mg/kg)+amlodipine(2.5 mg/kg) group. The model of two-kidney one clip re-nal hypertensive rats was established. The tail arterial blood pressure was detected once a week. After 20 weeks, rats were sacrificed and specimens were collected. The left ventricular mass index (LVMI) was assessed. The myocardial ultrastructur-al changes were observed by electron microscope. Values of plasma renin activity (PRA), angiotensionⅡ(AngⅡ) and atrial natriuretic peptide (ANP) were measured by enzyme linked immunosorbent assay (ELISA).Results Compared with sham group, the levels of systolic blood pressure (SBP), LVMI, PRA, AngⅡand ANP were significantly higher in 2KIC group (P＜0.01). Compared with 2KIC group, values of SBP, LVMI, PRA and ANP were significantly lower in telmisartan group and am-lodipine group (P＜0.01), but the value of AngⅡwas significantly higher (P＜0.01). The levels of SBP, LVMI, AngⅡand ANP were significantly lower in combined medication group than those of single drug medication group (P＜0.01). There was no significant difference in the plasma PRA level between those groups (P>0.05). Results of myocardial electron microsco-py showed that the left ventricular remodeling was significantly improved in combined treatment group. Conclusion Telmisartan and amlodipine can effectively improve the left ventricular remodeling induced by hypertension. There was more effective therapy using both medications together.

Objective:To explore the quality of inventory samples of a biobank stored in a deep freezer from 0 to over 10 years in Shanghai Ruijin Hospital. Methods:We extracted 24 pairs of stocked gastric cancer samples between 2003 and 2014. We used 1%aga-rose gel electrophoresis to analyze DNA and RNA purity and integrity while adding the RNA integrity number (RIN) for precise analysis. Bicinchonininc acid (BCA) assay was used for protein concentration evaluation. Coomassie brilliant blue method was used for protein integrity assay. Results: The samples were divided into four groups according to cryopreservation period (<2 years, 3-5 years, 6-8 years, and>9 years). No significant difference in DNA integrity was found between the groups (P>0.05);however, DNA degradation in normal gastric mucosa was faster than that in gastric cancer tissue (P=0.023). The RIN significantly declined when the storage period was 6 years or longer (P=0.018). No significant difference in protein concentration was observed between different groups. Using Coo-massie brilliant blue method, we found significant differences in preserved proteins with different molecular weights. Proteins with varying molecular weights were detected in the groups with the following cryopreservation periods:>9 years, a small number of low-molecular-weight (average 36.5 KD) proteins;6-8 years, medium-molecular-weight (average 65.63KD) proteins;3-5 years, high-molecu-lar-weight (average 127.5 KD) proteins;<2 years, high-molecular-weight (average 160 KD) proteins. Conclusion:Cryopreservation does not exert an obvious effect on DNA. If the cryopreservation period is more than 5 years, serious degradation of RNA should occur;like-wise, degradation of proteins with higher molecular weight should occur.

AIM: To study the effect of the overexpression of p16 on an anion exchange function of band 3 in HeLa cells. METHODS: The expression of p16 and band 3 in HeLa cells was detected by immunohistochemistry (IHC). The p16 cDNA was subcloned to plasmids pEGFP-C1 by PCR and identified by restriction enzyme digestion and sequencing, and then, the recombinant pEGFP-C1-p16 plasmids were transiently transfected into HeLa cells. The expression of fusion protein in HeLa cells was detected by fluorescence microscope. 6-methoxy-N-(3-sulfopropyl)-quinolinium(SPQ)fluorescent probes were used to detect the anion exchange function of band 3. RESULTS: P16 and band 3 were expressed in HeLa cells. The amplificated p16 cDNA sequence was the same as the report sequence. The transfective efficacy of pEGFP-C1-p16 was above 60%. The anion exchange function increased after the transfection of pEGFP-C1-p16 plasmids. CONCLUSION: p16 facilitates the anion exchange function of band 3 in HeLa cells.

OBJECTIVETo investigate the effect of topical application of leptin in promoting burn wound healing in rats.

METHODSFour parallel second-degree burn wounds induced on the back of 18 Wistar rats were divided into leptin treatment group (treated with topical application of 400 ng/ml leptin dissolved in PBS) and control group (treated with PBS). The time of wound healing was recorded, and the wound area that was not healed was measured at 3, 7, 11, 15, and 19 days after burns. The tissue at the peripheries of the wound was sampled at 7, 14 and 21 days after burns for pathological examination with HE staining and immunohistochemistry for proliferating cell nuclear antigen (PCNA) to evaluate the proliferation of keratinocytes.

RESULTSCompared with the control group, leptin-treated wounds showed a shorter time (by 2-3 days) of wound healing, and significant differences were found between the two groups in healing at 7, 11, 15, and 19 days after burns. HE staining and immunohistochemistry revealed a faster rate of epidermis growth and a greater thickness of the cuticular layer in leptin-treated wounds at 7, 14 and 21 days after burns. PCNA positivity in the keratinocytes was stronger in leptin-treated wounds than in the control wounds at 7 and 14 days, but no such distinct difference was noted at 21 days between the two groups.

CONCLUSIONTopical application of leptin can promote re-epithelization in burn wounds to shorten the wound healing time of burns.

Administration, Cutaneous ; Animals ; Burns ; drug therapy ; Leptin ; administration & dosage ; therapeutic use ; Male ; Rats ; Rats, Wistar ; Wound Healing

Objective To develop and evaluate a porcine model for laparoscopic ureterovesical reimplantation (LUR) training. Methods Ten female pigs with a mean weight of 30 kg were used and the animals were placed on supine position after anesthesia. One 10 mm port and two 5 mm ports were placed after the establishment of pneumoperitoneum. The horn of uterus was used as "ureter". A model simulating the performance of LUR was then established on the mini-pigs. Four trainees per-formed the LUR procedures on the animal models during an advanced laparoscopic urology training course, following the technique criteria exactly used in LUR. The learning curve was analyzed in terms of operation time. Results The porcine model for laparoscopic training was established suc-cessfully and 4 LUR trainings was performed on each porcine. Each trainee performed 10 LURs on the models during the training course. The operation time declined from 170±10 rain initially to 90±4 rain after the training course (P<0.01). At the end of this training, all trainees could accomplish a watertight LUR procedure on the model. Conclusions The establishment of the training model is feasible. The trainees could acquire skills needed to perform LUR in vivo based on this simple model and to develop dexterity and facility in laparoscopic manipulation of needles and sutures as well. The model provides a platform for basic techniques training of the ureteral reconstruction procedures.

In this study, about 350 bp cDNA fragment was amplified by PCR. After being sequenced, the AE1-c-end gene fragment was cloned into EcoR I-Pst I site of pGADT7 to form AD ends in the yeast two-hybrid system. The recombinant plasmid was transformed into yeast AH109, and the expression in the yeast was observed. The results demonstrate that AE1-c-end was obtained. pGADT7-AE1-c-end has no toxic effect on the yeast. It can serve as a target gene of yeast two-hybrid system.
Cloning, Molecular ; DNA, Complementary ; Plasmids ; Polymerase Chain Reaction ; Two-Hybrid System Techniques ; Yeasts