Objective To evaluate the laparoscopic Palomo procedure in the management of varicocele in adolescent by assessment of the postoperative testicular growth and the changes of the biggest vessel diameter of varix. Methods From July 1999 to March 2004, laparoscopic high mass - ligation of testicular vessels according to Palomo procedure was performed on 31 boys who all presented with left varicocele Before and after operation at 1 month,3 and 6 months, the biggest vessel diameter of varicocele was measured by ultrasonography, testicular blood supply was evaluated by color Doppler flow imaging(CDFI) ,and fastis was recorded in length, width and thickness by ultrasonographic measurement, which volume was calculated by the formula V =?/6 ? length ? width ? thickness. Also the ratio of left to right testicular volume was calculated, and the left testicle was thought atrophy if the ratio was below 75% The biggest vessel diameter of varix, testicular blood supply, and the postoperative testicular increasing volume were compared before and after operation. Results The biggest vessel diameter of varicocele significantly reduced 1 month after operation(P

Abstrate:Aim To construct HEK293 cell line with stable and high expression of sodium taurocholate cotransporting polypeptide (NTCP ) efficiently and rapidly.Method Vector expressing EGFP-NTCP fusion protein was constructed and verified by DNA sequencing.The pEGFP-NTCP expression vector was transfected into HEK293 cells by FuGENE 6 transfection reagent. The transfected cells with high expression of green fluorescent protein were selected using fluorescence microscope for screening of G418 for 14 days to obtain cell lines stably and highly expressing NTCP.NTCP expression was detected by RT-PCR,qRT-PCR, Western-blot and the uptake experiment of taurocholic acid.Re-sult RT-PCR,qRT-PCR,Western-blot and the uptake experi-ment revealed that compared to the control cells,the expression of NTCP was significantly positive (P<0.01)in stable trans-fected cells showing green fluorescence (P<0.05 ).Conclusion The HEK293 cell line with stable and high expression of NTCP has been established efficiently and rapidly,which provides a cellular model for the study of the mechanism of the uptake of bile acid derivatives.

5-10 years old were 33 cases;febrile children were 55 cases,persistent cough in 65 cases,among of those cases,the wheezing were 8 cases,nonproductive cough 45 cases,nasal discharge 7 cases in early right lung pneumonia in 38 cases,left lung pneumonia in 23 cases,pulmones involved at same time were 8 cases.Among of those children who senerated plenritis were 9 cases,pleural effussion 1 case;complication besides lung appeared 32 children.Complication of digestive and urinary system had 8 cases(11.6%),respectively.Complication of cardiovascular system had 3 cases(4.3%).Complication of nervous system and skin had 4 cases(5.8%),respectively.Complication of hematological system had 5 cases(7.2%).Eight cases(11.6%) showed damage of no more one organ,Fifty-nine cases first treated by erythromycin in vein,then by azithromycin in oral,10 cases were treated by azithromycin oral medication.Thirty-three cases fully recovered,36 cases improved.Conclusions SMPP are more founded in school children,the complication of besides lung are more serious than that of non-segmental pneumonia,it is security and relatively satisfaction to treat with erythromycin and azithromycin in sequential therapy.

Objective: To observe the clinical efficacy of herb-partitioned spreading moxibustion at Baliao points plus climen for diminished ovarian reserve (DOR). Methods: A total of 60 patients with DOR were randomized into a spreading moxibustion group and a Western medicine group by the random number table method, with 30 cases in each group. The Western medicine group was treated with climen, starting from the 5th day of the menstrual cycle for 21 d. The spreading moxibustion group was treated with herb-partitioned spreading moxibustion at Baliao points on the basis of the medication in the Western medicine group, 1 h per time, once a week. The treatment was performed for 1 month as one treatment course in both groups, for 3 courses in total. The serum follicle-stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E2) in the patients were measured before and after treatment. The peak systolic velocity (PSV) and resistance index (RI) were also detected. The traditional Chinese medicine (TCM) symptom score was evaluated. The clinical efficacy was evaluated after treatment. Results: The total effective rate in the spreading moxibustion group was 93.3％, which was significantly higher than 80.0％ in the Western medicine group, and the difference between the groups was statistically significant (P<0.05). After treatment, the TCM symptom scores, the serum FSH levels, FSH/LH ratios and RI in both groups decreased, and the intra-group differences were all statistically significant (all P<0.05). The serum E2 level and PSV increased compared with those in the same group before treatment, and the intra-group differences were statistically significant (all P<0.05). After treatment, the TCM symptom score, the serum FSH level, FSH/LH ratio and RI in the spreading moxibustion group were lower than those in the Western medicine group, while the serum E2 level and PSV were higher than those in the Western medicine group, and the differences between the groups were statistically significant (all P<0.05). Conclusion: Herb-partitioned spreading moxibustion at Baliao points plus climen can produce valid therapeutic efficacy for DOR. It can improve the clinical symptoms, regulate serum hormone levels and increase ovarian blood perfusion, thus improving ovarian reserve function, producing more significant efficacy than climen alone.

Aim To screen a more suitable transfection recep-tor,and improve the efficiency of constructing cell lines highly expressing human peptide transporters 1 (hPepT1 ).Methods The recombinant plasmid pcDNA3.1 (+)-hPepT1 was transfect-ed into MDCK cells and HeLa cells by LipofectamineTM 2000 transfection reagent,respectively.The monoclonal cells were se-lected and cultured.Expression of hPepT1 mRNA and protein were determined by qRT-PCR and Western blot,respectively. The uptake capacity of Glysar in transfected cells was examined. Results Compared with wild type cells,the expression of hPepT1 and the uptake of Glysar in transfected MDCK cells and HeLa cells significantly increased (P <0.05).Although the up-take of Glysar in HeLa cells was higher than that of MDCK cells,on the contrary,the expression of hPepT1 and the uptake of Glysar in MDCK-hPepT1 cells was higher than that of HeLa-hPepT1 cells.Conclusion MDCK cells may serve as a more suitable transfected receptor for the construction of a cellular model with high expression of hPepT1 ,which would make the construction of a cell model highly expressing hPepT1 more effi-cient.

Objective To investigate the expression of osteopontin and matrix metalloproteinase-9 (MMP9),and the relationship of osteopontin and MMP9 in malignant melanoma.Methods Expression of osteopontin and MMP9 was measured by immunohistochemical SP method in 23 patients with primary cuta- neous malignant melanoma,17 patients with metastatic melanoma and 20 patients with pigmented nevus. Results Osteopontin and MMP9 were expressed respectively in 87.5% and 75.0% of 40 malignant melanoma specimens,15.0% and 10.0% of 20 pigmented nevus specimens.The expression of both osteo- pontin and MMP9 was significantly higher (both P＜0.05) in malignant melanoma than in pigmented ne- vus.There was no correlation between the expression of osteopontin and MMP9,with age,sex,lymph node metastasis or location of lesions (P＞0.05).Twenty-nine cases were positive for both osteopontin and MMP9,4 negative for either osteopontin or MMP9.Conclusion Both osteopontin and MMP9 were over- expressed in malignant melanoma,but neither was related to lymph node metastasis.

Objective To explore the safety and feasibility of the "many people, many point, many cooperation (3M)" precise locating handling method in transferring general anesthesia patients according to the business process reengineering (BPR) theory. Methods A comparative analysis on 138 patients who had undergone general anaesthesia operation between October 2016 and January 2017 was performed. The research objects were divided into two groups randomly. The patients in observation group (n=69) was transferred by 3M method, while the patients in control group (n=69) was transferred by traditional method. The indicators of the two groups were compared, including transferring time, the number of people involved, the rate of tube slippage during transferring, the changes of heart rates and systolic blood pressure in one minutes before and after handling. Results The time used to handle the patient is much shorter in observation group (134.38±41.69) s than that in control group (201.71±49.34)s, with statistical difference (P<0.05). It needed four people to transfer in all cases of the observation group, while more than four people are needed in 9 cases of the control groups. There was no tube slippage case in the observation group, while 6 tube slippage cases in control group. The changes of heart rates and systolic blood pressure of patients within one minute before and after carrying are much higher in control group than in observation group,the differences were statistically significant (P<0.05). Conclusions In contrast to the traditional transferring method, 3M handling method is safer and more efficient for patients after general anaesthesia operation. It was also showed having less effect on patients heart rate and blood pressure. In conclusion,3M handling method is well worth clinical application.

Epilepsy is a chronic disease of transient brain dysfunction caused by imbalance between excitation and inhibition of central nervous system.Gut microbiota can affect the functional signals of the brain through brain-gut-microbiota axis.At the same time,brain signals can also affect microbial activities and gastrointestinal physiology.This article reviews the recent research progress on changes of intestinal microorganisms in epilepsy patients and possible mechanism of intestinal microorganisms affecting epilepsy.

OBJECTIVENuclear factor-kappa B (NF-kappaB) is a critical transcription factor governing the expression of many cytokines that are involved in the pathogenesis of inflammatory diseases, such as asthma and rheumatoid arthritis. Melatonin (MT), a relatively safe and potent antioxidant which has shown efficacy in several chronic inflammatory models, may inhibit the expression of NF-kappaB and therefore might have a therapeutic use in asthma. This study aimed at observing the effect of MT on the expression of NF-kappaB and airway inflammation in a rat model of bronchial asthma.

METHODSTwenty-four male Sprague-Dawley (SD) rats weighing 120 g to 170 g were randomly divided into three experimental groups (8 in each): (1) Asthmatic group: Rats were immunized on day 1 by intraperitoneal injection of 100 mg ovalbumin (OVA) in 1 ml of saline with 100 mg of alu minum hydroxide. From day 15 the animals were challenged with aerosolized OVA (1% in saline) for 20 minutes per day for 7 consecutive days. (2) MT group: OVA-sensitized rats were injected intraperitoneally with 10 mg/kg MT 30 minutes before each OVA challenge. (3) CONTROL GROUP: OVA for inhalation and MT for intraperitoneal injection was replaced with normal saline (NS). Airway responsiveness to aerosolized acetylcholine of 24 rats was detected six hours after the last challenge. Then the rats were lavaged and total and differentiated leukocytes counts in bronchoalveolar lavage fluid (BALF) were performed after staining with Wright-Giemsa staining. At the same time, levels of nitric oxide (NO) in BALF, inducible nitric oxide synthesis (iNOS) and constitute nitric oxide synthesis (cNOS) in the lung tissues were assessed with the use of nitrate reductase and chemical colorimetry, respectively. The expression of NF-kappaB in the lung tissues was observed by means of immunohistochemical staining.

RESULTS(1) After OVA challenge, there was a significant decrease in airway responsiveness, lymphocytes and eosinophils in BALF in MT group compared with asthmatic group (P < 0.01 respectively); (2) There was a significant decrease in amounts of NO(2)(-)/NO(3)(-) in the BALF and levels of iNOS in the lung tissues in MT group comparing with asthmatic group (P < 0.01 respectively); and the levels of iNOS in the lung tissues was correlated positively with NO(2)(-)/NO(3)(-) in the BALF (P < 0.01), but there were no significant differences in activity of cNOS in any of the groups analyzed. (3) There was a significant increase in expression of NF-kappaB in lung tissues in asthmatic group compared with the other groups (P < 0.01), and so was in MT group compared with control group (P < 0.05).

CONCLUSIONSMT could partially inhibit the expression of NF-kappaB and down-regulate the activity of iNOS in lung tissue, decrease the production of NO in BALF. These data suggest that the inhibitory effect of MT probably play a role in decreasing airway hyperresponsiveness and airway inflammation of asthmatic rats model.

Animals ; Antioxidants ; pharmacology ; Asthma ; drug therapy ; metabolism ; Bronchoalveolar Lavage Fluid ; cytology ; immunology ; Disease Models, Animal ; Lung ; chemistry ; pathology ; Male ; Melatonin ; pharmacology ; NF-kappa B ; analysis ; Nitric Oxide ; analysis ; Rats ; Rats, Sprague-Dawley

Based on the transcriptome database of Salvia miltiorrhiza, specific primers were designed to clone a full-length cDNA of ent-kaurene oxidase synthase (SmKOL) using the RACE strategy. ORF Finder was used to find the open reading frame of SmKOL cDNA, and ClustalW has been performed to analysis the multiple amino acid sequence alignment. Phylogenetic tree has been constructed using MEGA 5.1. The transcription level of SmKOL from the hairy roots induced by elicitor methyl jasmonate (MeJA) was qualifiedby real-time quantitative PCR. The full length of SmKOL cDNA was of 1 884 bp nucleotides encoding 519 amino acids. The molecular weight of the SmKOL protein was about 58.88 kDa with isoelectric point (pI) of 7.62. Results of real-time quantitative PCR analyses indicated that the level of SmKOL mRNA expression in hairy roots was increased by elicitor oMeJA, and reached maximum in 36 h. The full-length cDNA of SmKOL was cloned from S. miltiorrhiza hairy root, which provides a target gene for further studies of its function, gibberellin biosynthesis and regulation of secondary metabolites.
Amino Acid Sequence ; Cloning, Molecular ; Computational Biology ; methods ; Cytochrome P-450 Enzyme System ; chemistry ; genetics ; Models, Molecular ; Molecular Sequence Data ; Phylogeny ; Protein Structure, Tertiary ; Salvia miltiorrhiza ; enzymology