Objective:To optimize the extraction process of berberine hydrochloride from Sanhuang Diyu oil by Box-Benhnken re-sponse surface methodology. Methods:The extraction process was optimized with the size of each medicine ( X1 ) , the liquid-solid ra-tio ( X2 ) and the extraction time ( X3 ) as the independent variables and berberine hydrochloride extraction amount ( Y) as the depend-ent variable, and the effects of variables and their interactions on the extraction efficiency were studied. Results:The experiment results indicated that the optimal extraction conditions were as follows:the size of each medicine was 60 meshes, the liquid-solid ratio was 18, and the extraction time was 1. 5 h. Under the above conditions, 3 batches of berberine hydrochloride from Sanhuang Diyu oil were ex-tracted. The average extraction rate of berberine hydrochloride was (16.6 ±0.6) mg·g-1(n =3). Conclusion: The extraction process of Sanhuang Diyu oil optimized by Box-Behnken response surface methodology is simple, stable and predictable.

Multi-drug resistance of breast cancer remains a major obstacle for effective treatment,which involves many complicated mechanisms,including drug transport in the body,metabolism and drug targets.Recent researches find that the tradition Chinese medicine not only has good effects in improving the body resistance and general situation of patients and enhancing the effects of chemoradiotherapy,but also plays a vital roles in the muti-drug resistance reversal of breast cancer.

Objective:To establish a method for the determination of atorvastatin and amlodipine in compound atorvastatin calcium and amlodipine besylate tablets. Methods:The determination was performed on a Gemini-NX C18 column (250 mm × 4. 6 mm, 5μm) with the mobile phase consisting of 20 mmol·L-1 potassium dihydrogen phosphate solution ( adjusting pH to 4 with phosphoric acid)-acetonitrile-methanol (30 ∶10 ∶60) at a flow rate of 1. 0 ml·min-1 . The detection wavelength was set at 240 nm and the column tem-perature was 30℃. The injection volume was 20μl. Results:The linear range of atorvastatine and amlodipine was 0. 4-40. 0μg·ml-1 and 0. 2-20. 0 μg·ml-1, respectively. The mean recovery of atorvastatin and amlodipine was 100. 6% and 99. 7%, and RSD was 0. 92% and 0. 85% (n=9), respectively. Conclusion:The method is special, stable and reliable, and can be used for the content determination of compound atorvastatin calcium and amlodipine besylate tablets.

Lornoxicam is a new non-steroidal anti-inflammatory drug,it can inhibit the activity of cyclooxygenases and the immunologic injury afteroperation.Its efficacies on preoperative,operative period and postoperative analgesia are similar to those of opioid.Lornoxicam can decrease the dosage of opioid drugs and has the good qualities of short half-life,few adverse effects and satisfactory tolerance.Lornoxicam can provide rapid and stable analgesic effect in perioperative phase.

Objective:To prepare and characterize the PEGylated liposomes containing total saponins of Paris Polyphylla. Meth-ods:Using the size, PDI, zeta potential and encapsulation efficiency of the liposomes as the indicators, the influencing factors in the preparation were optimized. The particle size, PDI and zeta potential were studied by a Malvern Zetasizer, the morphology was ob-served under a TEM, and the stability was studied as well. Results:The particle size, PDI, zeta potential and encapsulation efficiency of the PEGylated liposomes was (109. 4 ± 32. 7) nm, (0. 171 ± 0. 036), ( -36. 7 ± 4. 5) mV and (93. 5 ± 3. 2) %, respectively. The liposomes were small spheres with smooth surface under the TEM. The long term stability studies showed that the liposomes were stable in 3 months after stored at 4℃. Conclusion:The preparation technology of the PEGylated liposomes containing total saponins of Paris Polyphylla is feasible, which can obtain liposomal preparations with high entrapment efficiency and good stability.

Objective:To optimize the ultrasonic-assisted extraction technology of polysaccharide from Tremella by Box-Benhnken response surface methodology .Methods:The single-factor extraction tests and response surface analysis were employed to optimize the technology conditions , including ultrasonic power , ultrasonic time and liquid/solid ratio.Results:The experiment results indicated that the optimal extraction conditions were ultrasonic power of 360 W, ultrasonic time of 23 min and liquid/solid ratio of 45 ∶1 ( ml· g-1 ) . Under the conditions , 6 batches of polysaccharide from Tremella were extracted .The average extraction rate of Tremella polysaccharides was 20.4%±1.8%(n=6).Conclusion: The ultrasonic-assisted extraction process of Tremella polysaccharide using Box-Behnken response surface methodology is simple with good predictability .

This study was aimed to evaluate the subchronic toxicity of diosgenin in mice. A total of 80 mice were divided into 4 groups, which were 0 (control), 100, 200, and 400 mg·kg-1 by the random number table. Intragastric administration was given once a day for 90 days in the assessment of subchronic toxicity of diosgenin among mice. The observed indexes contained body weight, fur color, diet, feces, and etc. The detected indexes contained blood routine analysis, blood biochemistry and pathological examination. The results showed that compared with the control group, the body weights of mice in the male medication group were slight reduced. There were no significant hematologic and pathologic abnormalities. It was concluded that the subchronic toxicity of diosgenin with no observed adverse effect dose level was more than 400 mg·kg-1. The oral administration was relatively safe.

Objective To investigate the value of serum ischemia-modified protein (IMA),hypersensitivity C reaction protein (hs-CRP),myoglobin (MYO)creatine kinase MB (CK-MB)and ultra-sensitivity troponin T (hs-cTnT)in the early diagnosis of patients with acute coronary syndrome(ACS).Methods Serum levels of IMA,hs-CRP,MYO,CK-MB and hs-cTnT were meaused in 94 patients with ACS[including 40 cases of unstable angina pectoris(UAP),20 cases of non-ST segment elevation myocardial in-farction(NSTEMI),non-Q wave myocardial infarction and 34 cases of ST segment elevation myocardial infarction(STEMI),Q wave myocardial infarction]and 99 cases of controls.The efficiency,sensitivity,specificity,negative and positive predictive values in the early diagnosis of ACS were compared among 5 kinds markers by the ROC curve.Results The detection results of serum IMA,hs-CRP,MYO,CK-MB and hs-cTnT had statistically significant differences between the UAP,NSTEMI and STEMI groups with the normal control group (P <0.05),serum IMA,hs-CRP,MYO,CK-MB and hs-cTnT had statistically significant differences between the UAP group with the NSTEMI group and between the UAP group with STEMI group (P <0.05),but the difference between NSTEMI and STEMI group group was not statistically significant (P >0.05).Conclusion In the patients with ACS,the multiple indicators combined detection can achieve the early diagnostic value.

Objective To investigate MR imaging features of parotid gland in Sj?gren′s syndrome ( SS).Methods Twenty-seven cases of xerostomia patients were collected and divided into SS group ( n=21) and non-SS group (n=6) according to the international classification (diagnosis) criteria for SS.Ten healthy volunteers were recruited as the control group.All the subjects underwent conventional MRI of parotid gland and MR sialography ( MRS).Standard deviation of T 1 WI and T2 WI signal intensity among 3 groups was observed, meanwhile, grading was made according to parotid glands , fat signal and parotid duct expansion degree respectively.With clinical diagnosis as the gold standard , diagnostic value of conventional MRI , MRS and their combination used in SS was compared.One-way ANOVA was used in comparison of standard deviation of parotid gland′s signal intensity among 3 groups , and Chi-square test was applied in comparison of conventional MRI and MRS diagnostic value.Moreover , Kappa value was calculated to assess the consistency of two grading results in SS.Results Signal intensity of parotid glands in control group and non-SS group was homogeneous.However , bilaterally diffused and heterogeneous high signal intensity on both T1WI and T2WI was found in SS patients, which was depressed on T2WI fat suppression sequences.Forty-two parotid glands were graded by fat signal:Grade 0 (n=2 glands), Grade 1 (n=10), Grade 2 (n=10), Grade 3 (n=6) and Grade 4 (n=14).Parotid peripheral ducts of control group and non-SS group were unexpanded , while bilaterally expanded parotid peripheral ducts were shown in SS patients.The grading of 42 parotid glands by expansion degree of parotid duct , Grade 0 was rated in 12, Grade 1 in 8, Grade 2 in 10, Grade 3 in 5, and Grade 4 in 7.Standard deviation of T1WI signal intensity of parotid glands among SS group , non-SS group and control group were 124.1 ±30.0, 81.8 ±27.6, and 86.3 ±35.0 respectively;and standard deviation of T 2 WI signal intensity were 115.1 ±35.2, 69.8 ±23.5, and 80.1 ±31.4 respectively; the standard deviation of T 1 WI and T2 WI signal intensity of SS group was higher than both non-SS group and control group′s ( F value =13.780 and 13.301, respectively, P <0.01), however, the difference of standard deviation of signal intensity of non-SS group and control group had no statistical significance (P>0.05).Among 42 parotid glands with SS, conventional MRI and MRS showed parotid gland lesions in 40 and 30 respectively , and the difference was statistically significant (χ2 =13.04, P=0.013).There was no false positive result.The combination of the two methods detected all 42 lesions.The consistency of detecting parotid abnormalities with SS between conventional MRI and MRS was poor (Kappa=0.12, P=0.092).Conclusions Diffuse fatty infiltration on conventional MRI and diffuse peripheral duct dilatation on MRS in the parotid gland are characteristic features of SS , and conventional MRI could be used as the preferred technique for the SS.combination with MRS may improve diagnostic accuracy.

To construct an HSV-1 vector vaccine carrying HIV-1 antigens, HIV-1 gp160, gag, protease and the expression elements were chained together, and then inserted into the internal inverted repeat sequence region of HSV-1 by bacterial artificial chromosome technology. Firstly, HIV-1 gp160 (including type B and C), gag and protease genes were cloned into pcDNA3 in series to generate the pcDNA/gBgp and pcDNA/gCgp, then the recombinant plasmids were transfected into 293FT cells, and HIV-1 antigen was detected from transfected cells by Western blotting. Then the expression cassettes from pcDNA/gBgp and pcDNA/gCgp, comprising HIV-1 antigen genes and expression elements, were cloned into pKO5/BN to generate the shuttle plasmids pKO5/BN/gBgp and pKO5/BN/gCgp. The shuttle plasmids were electroporated into E. coli cells that harbor an HSV-BAC, the recombinant bacteria were screened, and the recombinant DNA was extracted and transfected into Vero cells. The recombinant virus was purified through picking plaques, the virus' DNAs were identified by Southern blotting; HIV-1 antigen was detected from the recombinant HSV-1 infected cells by Western blotting, and the virus' replication competent was analyzed. As the results, gp160 and gag proteins were detected from 293FT cells transfected with pcDNA/gBgp and pcDNA/gCgp by Western blotting. The recombinant bacteria were generated from the E. coli electroporated with pKO5/BN/gBgp or pKO5/BN/gCgp. The recombinant HSV was purified from the Vero cells transfected with the recombinant DNA, the unique DNA fragment was detected from the genome of recombination HSV by Southern blotting; gp120 and gp41 were detected from the infected cells by Western blotting, and the recombinant HSV retained replication competent in mammalian cells. The results indicate that the recombinant HSV carrying HIV-1 gp160, gag and protease genes was generated, the virus retains replication competent in mammalian cells, and could be used as a replicated viral vector vaccine.
Animals ; Cercopithecus aethiops ; Chromosomes, Artificial, Bacterial ; DNA, Recombinant ; genetics ; DNA, Viral ; genetics ; Escherichia coli ; HIV Antigens ; genetics ; immunology ; HIV Envelope Protein gp160 ; genetics ; immunology ; HIV Protease ; genetics ; immunology ; Herpes Simplex Virus Vaccines ; immunology ; Herpesvirus 1, Human ; physiology ; Plasmids ; Transfection ; Vero Cells ; Virus Replication ; gag Gene Products, Human Immunodeficiency Virus ; genetics ; immunology