In this study, SD rats were orally administrated with oteracil potassium (300 mg . kg-1 . d-1 ) to prepare the hyperuricemia model, and divided into normal, model, Allopurinol, LE high dosage, middle dosage and low dose (200, 100, 50 mg . kg-1 . d-1) groups. The rats were orally administrated with test drugs 1 hour later after being orally administrated with Oteracil potassium. After 7 days, serum uric acid, serum creatinine, uric acid and expression of relevant transporters in kidney were tested to study the regulatory effect of leonurus extracts on serum uric acid, renal function and relevant transporters in kidney of rats with hyperuricemia. Compared with the model group, the leonurus extract group could significantly down-regulate serum uric acid and creatinine levels of rats with hyperuricemia, and increase the urine uric acid level. Meanwhile, leonurus extracts could notably down-regulate the mRNA expressions of urate transporter 1 (URAT1) and glucose transporter 9 (GLUT9), up-regulate the mRNA expressions of organic cation transportanter (OCT) and Carnitine transporter (OCTN) and promote the excretion of uric acid of kidney.
Allopurinol ; pharmacology ; Animals ; Blood Urea Nitrogen ; Creatinine ; blood ; Disease Models, Animal ; Down-Regulation ; Gene Expression Regulation ; drug effects ; Hyperuricemia ; blood ; drug therapy ; Kidney ; drug effects ; Leonurus ; chemistry ; Male ; Organic Anion Transporters ; genetics ; Oxonic Acid ; administration & dosage ; Plant Extracts ; isolation & purification ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Specific Pathogen-Free Organisms ; Up-Regulation ; Uric Acid ; blood

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.

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.

OBJECTIVETo evaluate the impact of recombinant human growth hormone (rhGH) combined with hypocaloric nutrition on nitrogen balance and blood glucose in patients after gastrointestinal operation.

METHODSForty-eight patients undergone abdominal operations were randomly divided into either subcutaneous rhGH (0.15 IU/kg) or placebo (menstruum) injection daily for 7 days postoperatively. These two groups had similar nutritional intakes. Body weight, blood cell count, liver and renal function, plasma albumin, prealbumin, transferrin and fibronectin were measured at day 1 before operation as baseline and day 3 and day 10 after operation by standard laboratory techniques. Nitrogen balance and blood glucose were measured from day 3 to day 9 after operation.

RESULTSAt postoperative day 10, the cumulative nitrogen balance and the level of plasma fibronectin were significantly higher in rhGH group [13.21 g,(104.77+/-19.94) mg/L] than those in placebo group [-6.88 g, (93.03+/-16.03) mg/L] (P<0.05). Furthermore, at day 4 and from day 6 to day 9 after operation, daily nitrogen balance was higher in rhGH group than that in control group (P<0.05). Mean blood glucose level was significantly higher in rhGH group from day 3 to day 6 after operation [(7.68+/-2.15) vs (5.95+/-2.34) mmol/L, P<0.05]. But hyperglycemia could be controlled well by insulin treatment.

CONCLUSIONGrowth hormone combined with hypocaloric nutrition is effective and safe in promoting positive nitrogen balance and protein synthesis in post-operative patients, which is also beneficial to the improvement of nutritional status and prognosis.

Adult ; Aged ; Blood Glucose ; metabolism ; Caloric Restriction ; Digestive System Surgical Procedures ; Female ; Human Growth Hormone ; therapeutic use ; Humans ; Male ; Middle Aged ; Nitrogen ; metabolism ; Nutritional Support ; methods ; Postoperative Period ; Recombinant Proteins ; therapeutic use

OBJECTIVETo characterize molecular and cytogenetic abnormalities in six 46, XX males, and to investigate the clinical manifestations and underlying mechanisms in such patients.

METHODSClinical data of six XX male patients were collected. Karyotyping, multiple polymerase chain reaction (PCR) and fluorescence in situ hybridization (FISH) were utilized to detect and locate the sex determining region (SRY) gene.

RESULTSPCR and FISH showed that all patients were SRY-positive XX males. All patients have their SRY gene located at the tip of derivative X chromosomes, which have resulted from translocation between short arms of X and Y chromosomes. High resolution karyotyping at 550-750 band level has revealed that the translocation breakpoints were at Xp22.33 and Yp11.2 in three patients. In the remaining patients, the breakpoints were either at Xp22.32 and Yp11.31 or Xp22.31 and Yp11.2. The breakpoints at Xp22.32, Xp22.31 and Yp11.31 were rarely reported. Genotype-phenotype correlation analysis indicated that the clinical manifestations were age-specific. Four adult patients have come to clinical attention due to infertility, with typical features including azoospermia and testis dysgenesis, whereas poorly developed secondary sexual characteristics and short stature were main complaints of adolescence patients, and short stature was the sole symptom in a child patient.

CONCLUSIONCombined karyotyping, PCR and FISH are important for the analysis of XX males. Particularly, high resolution karyotyping is valuable for the refinement of chromosome breakpoints and detailed analysis of genotype-phenotype correlation.

46, XX Disorders of Sex Development ; genetics ; Adolescent ; Adult ; Child, Preschool ; Chromosomes, Human, X ; Chromosomes, Human, Y ; Genetic Association Studies ; methods ; Humans ; Karyotyping ; methods ; Male ; Sex Chromosome Aberrations ; Translocation, Genetic ; Young Adult

AIMTo establish a simple method for molecular identification of original plants of D. chrysanthum and D. fimbriatum using molecular marker rDNA ITS region.

METHODSRestriction patterns of ITS fragments were obtained using PCR-RFLP method. The PCR products of D. chrysanthum and its morphologically allied species were digested at 37 degrees C by Cla I and Apa LI, those of D. fimbriatum and its morphologically allied species were digested by Sph I.

RESULTSD. chrysanthum, D. fimbriatum and their morphologically allied species could be identified by predicted restriction profiles of PCR-RFLP. The botanical origin of twenty-five fresh samples of "Shihu" collected in markets was identified by this method.

CONCLUSIONThe results showed that PCR-RFLP analysis of the rDNA ITS region is a feasible, simple and inexpensive method for determining the botanical origin of the traditional Chinese medicine "Shihu".

DNA, Plant ; analysis ; DNA, Ribosomal ; analysis ; Dendrobium ; classification ; genetics ; Drug Contamination ; Plants, Medicinal ; classification ; genetics ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Sequence Analysis, DNA ; Species Specificity

Bias ; Confounding Factors (Epidemiology) ; Epidemiologic Methods ; Humans

Objective To investigate HPV infection and its correlation with the expression of sur- vivin and proliferating cell nuclear antigen (PCNA) in condyloma acuminatum (CA).Methods HPV6/11 and 16/18 DNA were detected by PCR in 41 cases of CA and 23 specimens of normal skin.The expression of survivin and PCNA was measured by immunohistochemical technique in these two groups. Results The infection rates of HPV6/11 and 16/18 were 87.80% (36/41) and 19.51%(8/41),respective- ly,in CA,and 0 in normal skin epidermis.The expression of survivin and PCNA was found in 53.66% (22/41) and 87.80% (36/41) of CA ,and was 8.70%(2/23)and 47.83% (11/23) in normal skin,respec- tively.The expression of survivin was positively correlated with the high expression of PCNA in CA.The expression of survivin and PCNA was higher in HPV6/11 positive lesions than in HPV6/11 negative lesions. Conclusion HPV infection may up-regulate the expressions of survivin and PCNA,inhibit the apoptosis and promote the proliferation of keratinocytes.

Objective To observe the effect of isokinetic training of knee flexors and extensors on walking ability in patients with knee osteoarthritis(KOA). Methods From December,2016 to June,2017,120 patients with KOA in our hospital were randomly divided into blank group(n=40),control group(n=40)and observation group(n=40).All the groups were injected with sodium hyal-uronate intraluminally,and received joint mobilization.The control group added conventional strength training, including isotonic and isometric static squat exercise of muscles around the knee. The observation group re-ceived knee isokinetic training additionally,the training mode was ordinary isokinetic/centripetal mode.Before and four weeks after treatment, the peak torque (PT), the Lysholm score and Visual Analogue Scale (VAS) for pain,ten-meter walking time and six-minute walking distance were tested. Results After treatment,the PT value of knee flexors and extensors,the scores of VAS and Lysholm,ten-meter walking time and six-minute walking distance significantly improved in all groups(P<0.05),among which the observa-tion group was the best(P<0.05). Conclusion Isokinetic muscle strength training of knee flexors and extensors can improve the walking ability of patients with KOA.

OBJECTIVETo construct a cell model of 4.1R gene knockout in murine macrophage cell line RAW264.7 using CRISPR/Cas9 technique.

METHODSThree high?grade small?guide RNAs (sgRNAs) that could specifically identify 4.1R gene were synthesized and inserted into lentiCRISPRv2 plasmid. RAW264.7 cells were infected with sgRNA?Cas9 lentivirus from 293T cells transfected with the recombinant sgRNA?lentiCRISPRv2 plasmid, and the positive cells were screened using puromycin and the monoclonal cells were obtained. The expression of 4.1R protein in the monoclonal cells was measured by Western blotting, and the mutation site was confirmed by sequence analysis. Result A 4.1R gene knockout RAW264.7 cell line was obtained, which showed a 19?bp deletion mutation in the 4.1R gene sequence and obviously enhanced proliferation.

CONCLUSIONWe successfully constructed a 4.1R gene knockout macrophage cell line using CRISPR/Cas9 technique, which may facilitate further investigation of the function of 4.1R in macrophages.