Objective A HPLC method to determine the content of Baicalein in Tangtai capsule was established. Method The separation was performed by Zorbax C18 column with acetonitrile and 0.5% phosphorice acid. The elution program was 1~15 min, 15%～60% acetonitrile. The flow rate was 1 mL/min and column temperature was 25 ℃. Content was detected at a wavelength of 280 nm. Result The linear range was 0.936～18.72 ?g, regression:Y =4849.53X-0.9593 (r=0.9999). The average recovery rate was 98.29% and RSD was 0.97% (n =5). Conclusion The method is simple, accurate and reproductive, and can be used for determining Baicalein in Tangtai capsule.

Background The special pathological change of diabetic retinopathy(DR)is microvascular disorder.Angiopoietin-like protein 2(Ang-2)is a new protein associated with genesis of blood vessels.Quercetin has multiple pharmacological action,including improving the microcircularion and the permeability of blood capillary.However,the action mechanism of Ang-2 on DR was unclear.Objective The present study was to investigate the effects of quercetin on Ang-2 and its receptor Tie2 expression in retina with diabetes mellitus.Methods Sixty clean male Wistar rats were randomized into 7 groups and 10 rats for each group,and 10 rats served as blank control group.Streptozotocin of 35 mg/kg was intraperitoneally injected in 60 rats to establish the diabetic models.Quercetins encapsulated by liposome with the doses of 50,150 and 250 mg/(kg · d)(3-5 ml)were used to gavage in different groups of models for 12 weeks,and normal saline solution and calcium dobesilate were used at the same fashion as the negative control group and positive control group,respectively.Twelve weeks later,the animals were sacrificed and retinas were isolated.Expressions of Ang-2 protein and Tie2 mRNA in retinas were detected by ELISA and RT-PCR,respectively.The usage and rearing of the animals complied with the Regulations for the Administration of Affair Concerning Experimental Animals by State Sciences and Technology Commission.Results ELISA showed that the A450 of Ang-2 in 150 and 250 mg/(kg · d)quercetin groups was 0.796±0.057 and 0.842±0.043 respectively and was lower than that negative group(1.012±0.046),showing statistically significant differences(q =2.95,2.698,P＜0.05).RT-PCR assay showed that expression of Tie2 mRNA(Tie2 mRNA/GAPDH mRNA)in retinas was 0.712±0.092 and 0.821±0.087,presenting statistically significant differences in comparison with negative group(1.182±0.098)(q =3.497,2.852,P＜0.05).The expression levels of Ang-2 and Tie2 mRNA in retina were lowest in 150 mg/(kg · d)quercetin group.Conclusions Quercetin can improve the retinal microcirculation by downregulating the expressions of Ang-2 and its receptor in early period of diabetic rats.

This study aimed to investigate the drug susceptibility of wild-type and mutant avian influenza A (H7N9) virus neuraminidase (NA) to oseltamivir and zanamivir. Codon optimized DNA of H7N9 (A/ Hangzhou/1/2013) NA was synthesized and constructed into the pcDNA3.1/His vector (NA(H7N9-WT)). Mutant NA(H7N9-H274Y) and NA(H7N9-R292K) plasmids were constructed by directed mutagenesis PCR using NA(H7N9-WT) plasmid as the template followed by sequencing. NA plasmids were transfected into 293T cells and cell lysates containing NAs were collected 48 h post-transfection. Wild-type and mutant NAs were analyzed by Western blotting and their activities were tested by the 4-MUNANA-based assay. All three NAs were expressed and enzymatic activities were confirmed. The effects of oseltamivir and zanamivir on all three NAs were then tested. It showed that the half maximal inhibitory concentrations (IC50s) of oseltamivir carboxylate on NA(H7N9-WT), NA(H7N9-H274Y) and NA(H7N9-R292K) were 1.6 nM, 15.1 nM, and > 1 000 nM with fold changes of 9 and > 625, respectively. The IC50 values of zanamivir on NA(H7N9-WT), NA(H7N9-H274Y), and NA(H7N9-R292K) were 1.1 nM, 1.4 nM, and 38.0 nM with fold changes of 1.3 and 34, respectively. These results indicated that oseltamivir and zanamivir could significantly inhibit NA(H7N9-WT). NA(H7N9-R292K) showed high-level resistance to both drugs (34-fold and 625-fold) and NA(H7N9-H274Y) was sensitive to both (1.3-fold and 9-fold). These results indicated that both oseltamivir and zanamivir could be used for patients infected with the H7N9 virus. However, when patients carried the H7N9 virus with a NA R292K mutation, other medications would be preferred over oseltamivir or zanamivir.
Antiviral Agents ; pharmacology ; Humans ; Influenza A Virus, H7N9 Subtype ; drug effects ; enzymology ; genetics ; Influenza, Human ; virology ; Microbial Sensitivity Tests ; Mutation ; Neuraminidase ; antagonists & inhibitors ; genetics ; metabolism ; Oseltamivir ; pharmacology ; Viral Proteins ; antagonists & inhibitors ; genetics ; metabolism ; Zanamivir ; pharmacology

OBJECTIVETo investigate clinical effects of arthroscopy in the treatment of medial meniscal cyst.

METHODSFrom June 2011 to January 2013, 7 patients with medial meniscal cyst were treated with arthroscopy. There were 3 males and 4 females,ranging in age from 27 to 63 years old,with a mean age of (43.93±2.10) years old. The cysts have been discovered for 3 to 30 months,with a mean time of (10.6±1.3) months. All the patients complained of knee pain,especially in the medial joint gap. The Pisani sign, Caklin sign and medial McMurray sign were all positive. Preoperative MRI examination confirmed the diagnosis. Lysholm score changes and clinical efficacy were observed through a six-month follow-up.

RESULTSThe postoperative Lysholm scores were all significantly higher than the preoperative scores. According to Sarimo standard, 6 patients got an excellent result, and 1 good.

CONCLUSIONArthroscopic treatment of medial meniscal cyst has replaced the traditional method, which could retain the normal meniscus as much as possible and repair the meniscus injury simultaneously, as well as get a good curative effect and a good recovery of knee function. This method is worthy of clinical application.

Adult ; Arthroscopy ; methods ; Cysts ; physiopathology ; surgery ; Female ; Humans ; Male ; Menisci, Tibial ; physiopathology ; surgery ; Middle Aged

Objective To investigate the expression of macrophage migration inhibitory factor (MIF) and CD_(74), the receptor of MIF, in preeclamptic placenta and its correlation with the pathogenesis of preeclampsia.Methods From March 2008 to November 2008,69 preeclamptic women who delivered in the Department of Obstetrics, Affiliated Hospital of Qingdao University Medical College,were recruited,including 33 women with mild preeclampsia (MPE group) and 36 women with severe preeclampsia (SPE group).Another 43 healthy pregnant women were taken as control group.Immunoturbidimetry was applied to measure the concentrations of C-reactive protein (CRP) in maternal blood.The expressions of MIF and CD_(74) in placenta were tested with immunohistochemistry and the expressions of MIF mRNA and CD_(74) mRNA were detected by semiquantitative RT-PCR.The relationship between maternal blood level of CRP and MIF mRNA and CD_(74) mRNA in placenta was analyzed in the MPE and SPE group.Results (1) MIF and CD_(74) were expressed in the placenta of all pregnant women in the 3 groups, as shown in brown-yellow color, and significantly higher expression was found in the MPE and SPE group.(2) The expression of MIF mRNA and CD_(74) mRNA in the MPE group (0.70±0.13 and 0.96±0.16), SPE group (0.88 ± 0.12 and 1.08 ± 0.15) were significantly higher than in the control group (0.67 ± 0.11 and 0.83 ± 0.14) (P < 0.01), and statistical significance was also found between the MPE and SPE group (P <0.01).(3)The maternal blood concentrations of CRP in the MPE and SPE group were significantly higher than in the control group [(15.3±7.0) mg/L and (21.6±9.1)mg/L vs (4.8 ± 1.8) mg/L, P <0.01] , and significant difference was also found between the MPE and SPE group (P <0.01).(4) In the two preeclamptic groups, the blood concentrations of CRP were positively correlated with the expression of both MIF mRNA(r =0.67 ,P <0.01)and CD_(74) mRNA(r =0.83 ,P <0.01) in placenta.Positive correlation was also found between the levels of MIF mRNA and CD_(74) mRNA in placenta (r =0.93 ,P < 0.01).Conclusions Overexpression of MIF and CD_(74) in the placenta may up-regulate the CRP level in maternal blood, resulting in systemic inflammatory reaction and vascular endothelium damage which may be involved in the pathogenesis of preeclampsia.

Objective To investigate the clinical application value of ductoscopic flushing in the treatment of early lactation acute mastitis.Methods 52 patients with early acute mammitis were divided into observation group (27 cases)and control group(25 cases)according to the principle of completely random.In the observation group, the patients were checked under the intervention of mammary duct firstly,and then the disease regions of the breast were flushed using ductoscope.In the control group,the patients were treated with artificial breast -milk.The two groups were all treated with the same antibiotics.The curative effect of the two groups was observed,and the statistical analysis was performed.Results In the observation group,the mass extinction time,pain relief time,pyretolysis time, hemogram recovery time,contralateral continued breast -feeding proportion,the proportion of abscess formation,the proportion of back -milk,the proportion of ipsilateral quadrant recurrence were (3.5 ±1.2)h,(5.0 ±0.9)h, (1.0 ±0.1)d,(1.0 ±0.3)d,92.6%,7.4%,7.4%,0.0% respectively,those in the control group were (24.0 ± 3.2)h,(2.0 ±2.1)h,(2.0 ±0.2)d,(3.0 ±0.3)d,88.0%,12.0%,12.0%,8.0% respectively.The differences between the two groups were statistically significant(t =1.72,0.36,0.43,0.72,χ2 =1.83,2.02,1.56,0.34,all P <0.05).Conclusion Ductoscopic flushing has good effect in the treatment of early lactation acute mastitis,and it is worthy of clinical promotion.

Aim To investigate whether rosuvastatin induced reduction of atherosclerotic plaque was related to the expression of Sialyltransferase ( ST6 Gal-Ⅰ) in ApoE-/ - mice. Methods Six-weeks old ApoE-/ -mice fed with high fat were divided randomly into three groups: baseline group ( n=12 ) , control group ( n=12 ) and rosuvastatin group ( n =12 ) . Sixteen weeks later, control group was sacrificed. Serum and aortic intima were saved. Control group and rosuvastatin group were fed for seven weeks continually. Concentra-tions of serum lipids(TC, TG, LDL and HDL) were analyzed. Sections from the aortic root were examined by Hematoxylin-Eosin( HE) staining. The size of ath-erosclerotic lesion in each section was evaluated. Ex-pression of ST6 Gal-Ⅰ in aortic intima was detected by immunohistochemistry. Results Plasma TG and LDL-C, plaque areas and intimal thickness of control group were significant higher than those of baseline group ( P<0. 05 ) . Those results indicated that the AS model was successfully constructed. After seven weeks, the plaque areas and concentrations of serum lipids of rosu-vastatin group were obviously smaller than those of con-trol group(P<0. 05). The expression of ST6Gal-Ⅰin aortic root was decreased in control group compared to the baseline, and which was increased in control group compared to the rosuvastatin group. Conclusion Ro-suvastatin could inhibit the progression of atherosclero-sis, which might be related to the expression of ST6Gal-Ⅰ in aortic root.

60 years old)was significantly higher (41)than that in patients

Objective To study the effects of shRNA-CtBP2 on the growth of prostate cancer PC3 cells. Methods There were three experimental groups in this study,which include blank control group, empty plasmid transfected group and transfected shRNA group. CtBP2 mRNA sequence is targeted by 3 pairs of designed interfering shRNA to built shRNA-Ct-BP2 recombinant plasmid then it is transfected into PC3 cells. Reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot assays were used to detect the transcription and expression levels of CtBP2 mRNA and protein, respective-ly. PC3l proliferation was measured by MTT assay. Results Builting shRNA-CtBP2 recombinant plasmid and transfect-ing PC3 cells were successful. Transcription and expression levels of CtBP2 mRNA and protein were significantly decreased in shRNA-CtBP2 transfected PC3 cells. After CtBP2 silencing, cell proliferation was blocked in the shRNA-CtBP2 cells compared to that of blank control group (P<0.01). Conclusion shRNA-CtBP2 could significantly inhibit CtBP2 expres-sion, suppress the growth of PC3 cells, which suggests that CtBP2 may be a new target for PCa gene therapy.