To explore the feasibility of stable expression of Hantavirus H8205 strain G1 segment and human IL-2 fusion gene in Vero cells, and to examine the immune protection effects on mice vaccinated with this recombinant eukaryotic expression vector containing Hantavirus G1 gene and IL-2 gene. With the help of lipofectamine, the Vero cells were transfected with pcDNA3.1/HisB-IL-2-G1 and the positive cells were selected by G418. IFAT and SDS-PAGE electrophoresis were used to determine the stable transfection and expression of recombinant protein. Each mouse was inoculated with plasmids intramuscularly (i.m.) three times, 2 boosts were given at 2-week intervals, serum anti-hantavirus antibodies were detected by ELISA and neutralizing antibodies (NAb) were detected by Plaque Reduction Neutralization Test. The fusion protein expressed in Vero cells was 78 kD, corresponding to the estimated molecular size. The neutralizing antibody titers of mice with pcDNA3.1/HisB-IL-2-G1 were 1:20-1:80. IL-2/G1 fusion gene could be transferred in Vero cells and stably express the fusion protein. Specific humeral immune responses in mice can be induced with the recombinant eukaryotic expression vector containing the fusion gene, which lays the foundation for further development of therapeutic HTNV vaccine.

Objective To explore the acute toxicity and LD 50 of Tripterygium Hypoglaucum (Levl) Hutch(THH) solution to provide information for safe clinical application. Methods After oral administration of THH solution in mice, the mortality and the physiological and pathological changes were observed. Results The LD 50 (95% confidence limit) of THH in male and female mice was 79 g/kg(69～89 g/kg) and 100 g/kg (90～112 g/kg), respectively. No marked pathological change of the organs was found. Conclusion According to the standard of grading of acute toxicity, THH solution belongs to the moderate class. Therefore, it is safe in clinical practice and has a wide application.

Object To investigate the saponin from the seeds of Trigonella foenum graecum Linn (STFG) Methods The total saponin from STFG was extracted and purified by using the absorptive resin, the single saponin was isolated by using the column chromatography as well as dry column chromatography of silica gel H The chemical structure was elucidated by 13 CNMR , FAB MS, DEPT spectroscopic evidence and the results of fraction hydrolysis of acquiring their secondary glucosides Results A new saponin A from the total saponin has been obtained, the fraction hydrolysis carried out and the secondary glucoside Ⅰ and Ⅱ identified by determining the structure of saponin A The chemical structure of saponin A is: diosgenin 3 O ? L rhamnopyranosyl(1→4) ? D glucopyranosyl(1→4) ? D glucopyranoside The secondary glucoside Ⅰ is: diosgenin 3 O ? D glucopyranoside; Ⅱ is: diosgenin 3 O ? D glucopyranosyl(1→4) ? D glucopyranoside Conclusion Glucoside A is a new saponin with three molecules of sugar

Object To do detail investigation of the saponins from the Chinese materia medica Huluba (the seeds of Trigonella foenum graecum L ) Methods The pure saponins from the total saponins were isolated by employing the column chromatography and dry column chromatography of silica gel H Their chemical structures were elucidated by 13 C NMR , FAB MS, DEPT spectroscopic evidence and the results of the fraction hydrolysis of acquiring their secondary glucosides were obtained Results Two new saponins B and C were isolated and both were the glucosides consisted by four molecules of sugar with diosgenin The chemical structure of B is: diosgenin 3 O ? L rhamnopyranosyl (1→3) ? L rhamnopyranosyl (1→4) ? D glucopyranosyl (1→4) ? D glucopyranoside And saponin C is: diosgenin 3 O ? D glucopyranosyl (1→4) ? L rhamnopyranosyl (1→4) ? D glucopyranosyl (1→4) ? D glucopyranoside Conclusion Saponins B and C are two new ones with four molecules of sugar respectively

Objective To investigate the saponins from the Chinese materia medica Huluba (the seeds of Trigonella foenum-graecum L.). Methods To isolate and purify the single saponin by the column chromatography and dry column chromatography of silica gel H. The structural elucidation was carried on by spectroscopic evidence of (()~(13)C-NMR), FAB-MS, DEPT and the results of the fraction-hydrolysis of acquiring the secondary glucosides. Results The newly isolated saponin D was consisted by six molecules of sugar with diosgenin. The chemical structure of saponin D is: diosgenin-3-O-?-L-rhamnopyranosyl (1→3)-?-D-glucopyranosyl (1→4)-?-L-rhamnopyranosyl [(1→3)-?-L-rhamnopyranosyl] (1→4)-?-D-glucopyranosyl (1→4)-?-D-glucopyranoside. Conclusion Saponin D is a new one consisted of six molecules of su-(gar) with diosgenin.

Four triterpenoids were isolated and purified from the 95% ethanol extract of Maytenus guangxiensis by silica gel column chromatography, Sephadex LH-20 column chromatography, MCI column chromatography and preparative RP-HPLC. Their structures were determined from their physicochemical properties and spectral data. They were identified as maytguanone A (1), maytguanone B (2), 11α-methoxyurs-12-ene-1β,3β-diol (3), lup-20(29)-ene-3β,11α-diol (4). Compounds 1 and 2 are new triterpenoids, along with compounds 3 and 4 were isolated from M. guangxiensis for the first time. The cytotoxicity of compounds 1, 3 and 4 was evaluated using the MTT procedure with three cancer cell lines. The results show that compound 3 displayed good inhibitory effects against HeLa, with an IC₅₀ of 10.68 μmol·L^-1.

Objective To evaluate the accuracy and value of high-frequency ultrasound for breast cancer screening in Asian women. Methods Published studies on high-frequency ultrasound for screening breast cancer in Asian women were systemically searched and assessed by the Quality Assessment of Diagnostic Accuracy Studies (QUADAS) tool. Meta-DiSc 1.4 software was used for extracting data, calculating the summary sensitivity and specificity, and drawing the Summary Receiver Operating Characteristic (SROC) curve. Furthermore, the proportion of screening of diagnosis on early breast cancer (TNM stage 0, Ⅰ and Ⅱ ) by high-frequency ultrasound was calculated. Results Seven screening studies including 22 244 women were selected for Metaanalysis. According to the QUADAS items, 5 studies were classified as A degree and 2 studies were evaluated as B degree. For study in the heterogeneous of these 7 studies (Q=38.97, P＜0.0001 ),Random Effects Model (REM) was selected. The combined sensitivity (95%CI) and specificity (95%CI) were 0.785 (0.726-0.837) and 0.975 (0.973-0.977) respectively. The Area Under the Curve (AUC) of SROC was 0.9800. Among the follow-up studies of following period over one year, 96.9%of the diagnosed patients with breast cancer were at clinical stage Ⅱ or prior to it. Conclusion Because of its high accuracy, high- frequency ultrasound could be recommended for screening breast cancer in Asian women.

OBJECTIVETo explore the clinical and function efficacy of the treatment of Herbert B2-type scaphoid fracture through volar approach with double screws prograde internal fixation.

METHODSFrom December 2005 to June 2010,18 patients with Herbert B2-type scaphoid fracture were treated with double screws prograde internal fixation through volar approach, including 14 males and 4 females with an average age of 33.11 years (ranged from 21 to 52 years). The fracture healing situation was evaluated by X-ray examination. Functional outcome was assessed by using Modified Mayo wrist score.

RESULTSAll patients were followed up with a mean time of (25.06 +/- 4.00) months. The bone healing time was (3.55 +/- 0.65) months. The pain,passive range of motion, grip of wrist were improved at 4 months after operation (P < 0.05). The Mayo wrist score were improved from preoperative 42.78 +/- 7.32 at 4 months after operation (93.89 +/- 5.83) (P < 0.05), the results were excellent in 9 cases and good in 9 cases.

CONCLUSIONVolar approach with double screws prograde internal fixation is an efficacy and safety surgical technique for Herbert B2-type scaphoid fracture.

Adult ; Bone Screws ; Female ; Fracture Fixation, Internal ; instrumentation ; methods ; Humans ; Male ; Middle Aged ; Palmar Plate ; surgery ; Range of Motion, Articular ; Scaphoid Bone ; injuries ; surgery ; Wrist Injuries ; physiopathology ; surgery ; Young Adult

A UPLC-ESI-MS/MS method was used to determinate the main active fractions gallic acid, protocatechuic acid, myricetrin, hyperoside and quercitrin in Polygonum capitatum extracts by in situ intestinal perfusion models; the absorption rate constants and cumulative penetration rate of absorption were calculated. The effect of different drug concentrations, different intestine segments, bile and P-gp inhibitors on the absorption mechanism of Gallic acid and other compositions in P. capitatum extracts. The experimental results showed that gallic acid, protocatechuic acid, myricetrin and quercitrin were observed saturated at high concentration (P < 0.05). Bile had significant inhibition effect on protocatechuic acid absorption and had promotion effect on myricetrin and hyperoside absorption (P < 0.05). P-gp inhibitor verapamil could significantly enhance the absorption of Protocatechuic acid (P < 0.05). The overall trend for absorption of various compositions was that small intestine > colon. This indicated that the absorption mechanism of P. capitatum extracts in rat intestine was in line with fist-order kinetics characteristics. The composition could be absorbed in all of the different intestinal segments, and the absorption was mainly concentrated in small intestine. The protocatechuic acid may be the substrate of P-gp.
Animals ; Drugs, Chinese Herbal ; chemistry ; metabolism ; Female ; Intestinal Absorption ; Intestines ; chemistry ; metabolism ; Male ; Polygonum ; metabolism ; Rats ; Rats, Sprague-Dawley

AIM: To prepare and identify mouse polyclonal antibody against protein Hlb, which is the variant of major subunit of human ASGPR. METHODS: Hlb specific peptide was synthesized and coupled with keyhole limpet hemocyanin (KLH) for immunization. Then H1b-KLH conjugation was injected into mouse subcutaneously to produce polyclonal antibody. ELISA assay was used to detect the titer of the antibody. Antibody was also identified by Western blot and immunohistochemistry assays. RESULTS: Mouse antibody against Hlb was prepared after injection of H1bKLH conjugation. The titer of H1b antibody was about 1:10~5.Western blot confirmed its high specificity. This antibody could also be used for immunohistochemistry analysis. CONCLUSION: The successful preparation of the polyclonal antibody against protein H1b, which can discriminate the two variants of the major subunit of ASGPR with high specificity, will provide an efficient reagent for further study of the physiologic functions of H1b and its role in the pathogenesis of human disease.