Objective: To evaluate the safety and efficacy of nonmyeloablative preconditioning allogeneic hematopoietic stem cell transplantation (NMHSCT) in the therapy of unidentified relapse and primary solid tu-mors, and to study the anti-tumor immunity induced by the effect of Graft-Verus-Tumor (GVT). Methods: A to-tal of 13 difficult-to-treat cancer patients received NMSCT and the efficacy and side effects were observed. Re-sults: One case had CR, 2 cases had PR, 4 cases had SD, 5 cases had PD, and 1 case died of complica-tions associated with transplantation. One case was GVT (+++), 3 cases were GVT (++), 5 cases were GVT(+), and 4 cases were GVT (-). The main side effect was acute GVHD presented as diarrhea and infec-tion. VOD and brain disease were rare. Conclusion: Nonmyeloablative allogeneic stem cell transplantation is safe and effective for solide tumors.

Objective:To study the in vitro antimicrobial activity of traditional Chinese medicine herbal pair of euphorbia humifusa Willd. and portulaca oleracea L. . Methods: The antibacterial effect in vitro of the traditional Chinese medicine herbal pair and the single herb of euphorbia humifusa Willd. and portulaca oleracea L. was studied on staphylococcus aureus, escherichia coli and pseudo-monas aeruginosa by the 2-fold dilution method and the broth micro-dilution method in a 96-well plate. The minimum inhibitory con-centration ( MIC) , minimum bactericidal concentration ( MBC) and the diameter of inhibition zone were determined. Results:The ex-perimental strains showed the different sensitivity among the traditional Chinese medicine herbal pair and the single herb. The antibac-terial and bactericidal activity of the traditional Chinese medicine herbal pair was the most obvious(P<0. 05). As the temperature in-creasing, the antibacterial activity of all water extract on different experimental strains changed. The results of MIC and MBC showed that the effects of water extract on escherichia coli were strongest, that of stapphylococcus aureus were secong, ant that of pseudomonas aeruginosa were relatively weak. Conclusion:The traditional Chinese medicine herbal pair of euphorbia humifusa Willd. and portulaca oleracea L. has antibacterial and bactericidal activity in varying degrees on the experimental strains with some differences, and the changes in the application forms of traditional Chinese medicines has great influence on the antibacterial and bactericidal ability.

Aim To explore inhibitory effects and mechanism of engineering endostatin on growth and metastasis of melanoma cells in mice. Methods Melanoma cells(2× 106/mouse)were inoculated sabcutaneously to C57BL/6 mice. After tumorigenesis,endostatin(8mg/kg.d)was administrated to tumor-bearing mice,once a day ,twenty-one in all.Dietetic state and weight change of the tumor-bearing mice were observed and tumorous sige was measured during administration of endostatin. On 26thday,the tumor-bearing mice were sacrificed,subcutaneously tumorous weight was weighed and brain,lung ,liver,spleen and kidney were excised and sections were made to supply the pathological examination. Results Area under curve in the endostatin-treated group was obviously less than that in tamor control group(P∨ 0.01). Pathological study revealed that lavge areal necrosis arose in tumor and newborn cappillaries around the tumor disapeared. Conclusion Endostatin possosses strongly inhibitory effects on growth and metastasis of mouse melanoma and formation of newborn capillaries around tumor.

Objective To evaluate efficacy and safety of locally-produced pazufloxacin mesilate sodium chloride injection in the treatment of bacterial infections of respiratory and urinary tract.Methods A multi-center double-blind randomized controlled clinical trial was carried out to evaluate efficacy and safety of pazufloxacin mesilate sodium chloride in treatment for acute bacterial infection, as compared to those of levofloxacin hydrochloride and glucose injection as control treatment.A total of 244 patients with acute bacterial infection of respiratory and urinary tract were enrolled in the studies.120 in trial group and 120 in control group, with four withdrawals.Pazufloxacin mesilate and levofloxacin were administered intravenously by drip at a dose of 300 mg and 200 mg, every 12 hours for 7 to 14 days for trial and control groups, respectively.Resuits Overall efficacy of pazufloxacin mesilate was 77.0 percent and 93.5 percent in treatment for acute bacterial infections of respiratory and urinary tract.respectively, and that of levofloxacin was 80.6 percent and 89.6 percent, respectively.Overall bacterial clearance rate WaS 91.5 percent for pazufloxacin mesilate, 89.6 percent for respiratory tract infection and 94.1 percent for urinary tract infection, respectively.and 93.4 percent for levofloxacin, 97.3 percent for respiratory tract infection and 89.7 percent for urinary tract infection, respectively.No significant difference in adverse drug reactions between the two groups(P＞0.05)was found, with 4.88 percent and 7.44 percent for trial and control groups, respectively.Conclusions Pazufloxacin mesilate sodium chloride injection produced locally is a safe and effective antibiotic in treatment for acute infections of respiratory and urinary tract.

Anoectochilus, the fresh or dry whole grass of Anoectochilus roxburghii (Wall.) Lindl., has the effects of cooling blood, dispelling wind, dispelling dampness and resolving toxin, resulting in a wide range of clinical applications and large market demand. However, in the planting process, there are problems such as irregular planting management, excessive addition of pesticides, irregular processing links, even the phenomenon of selling adulteration, which seriously affects the quality of the medicine. In order to guarantee the quality and productivity of Chinese herbal medicines, this paper developed the standard operating procedures for the non-pollution Anoectochilus, including the non-pollution cultivation environment of Anoectochilus, planting methods, field management, pest control and quality control, etc., which provides guidance to promote the healthy development of the Anoectochilus industry.

Here, we presented a method to bacterially express the major structural protein L1 of Human Papillomavirus type 18 (HPV18) as soluble form. We found that the purified L1 could self-assemble to virus-like particles (VLPs). Further, we investigated the immunogenicity and the induced level of neutralizing antibody using these VLPs. First, the genome of HPV18 was cloned from a patient in Xiamen. It was used as template for PCR amplification of HPV18 L1 gene. The resultant DNA fragment was inserted into expression vector pTrxFus and expressed in Escherichia coli GI724. Second, L1 protein was purified by ammonium sulfate precipitation, ion-exchange chromatography and hydrophobic interaction chromatography; and the purified L1 was subjected to self-assembly to form VLPs with the removal of premixed reductant DTT. Finally, the size and morphology of these VLPs was investigated by Dynamic Light Scattering and Transmission Electronic Microscopy as 29.34 nm in hydrated radius and globular particles similar with native HPV18. The half effective dosage (ED50) and maximum level of neutralizing antibody elicitation were measured by vaccinations on mice, rabbit and goat using pseudovirus neutralization cell model. The results showed that the ED50 of HPV18 VLPs is 0.006 microg in mice, and the maximum titer of neutralizing antibody elicited in rabbit and goat is up to 10(7). As a conclusion, we can provide HPV18 VLPs with highly immunogenicity from prokaryote expression system, which may pave a new way for research and development of prophylactic vaccine for HPV18.
Animals ; Capsid Proteins ; biosynthesis ; genetics ; immunology ; Escherichia coli ; genetics ; metabolism ; Goats ; Human papillomavirus 18 ; immunology ; isolation & purification ; Mice ; Oncogene Proteins, Viral ; biosynthesis ; genetics ; immunology ; Rabbits ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Virion ; genetics ; immunology

We produced high pathogenic avian influenza H5N1 haemagglutinin protein HA1 in recombinant Pichia pastoris in a 10 L fermentor, to establish a high-density cell fermentation method. We studied the effects of different factors such as culture temperature, induced temperature, methanol feeding methods, trace elements on the growth of Pichia pastoris, the yield and the biologic activity of recombinant HA1 protein. The culture temperature in pre-induced and induced stage were optimized at 25 degrees C to adapt cell growth and recombinant protein expression, and induced temperature at 25 degrees C also resulted in higher biologic activity of rHA1 than at 30 degrees C. The binding activity of rHA1 against a wide-spectrum neutralizing antibody was susceptible to the presence of any trace elements, although trace elements would essentially benefit for the cell fermentation. As a conclusion, the expression level of rHA1 produced with optimized fermentation process reached 120 mg/L, which was 10.5 times higher than the one produced in regular shaking flask. The resultant high-density cell fermentation can likely produce rHA1 of H5N1 in large scale.
Fermentation ; Hemagglutinins, Viral ; biosynthesis ; genetics ; Influenza A Virus, H5N1 Subtype ; genetics ; metabolism ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis

As self-subunit swapping chaperones or metallochaperones, the activators assist nitrile hydratases to take up metal ions and they are essential for active expression of nitrile hydratases. Compared with nitrile hydratases, the activators have a low sequence identity. Study of the activation characteristics and the relationships between structures and functions of the activators is of great significance for understanding the maturation mechanism of nitrile hydratase. We co-expressed low-molecular-mass nitrile hydratase (L-NHase) from Rhodococcus rhodochrous J1 with four heterologous activators respectively and determined their activation abilities. Then we made sequence analysis and structure modelling, and studied the functions of the important domains of the activators. Results showed that all four heterologous activators could activate L-NHase, however, the specific activities of L-NHases were different after activation. L-NHase showed the highest specific activity after being activated by activator A, which was 97.79% of that of the original enzyme, but the specific activity of L-NHase after being activated by activator G was only 23.94% of that of the original enzyme. Activator E and activator G had conserved domains (TIGR03889), and deletion of their partial sequences resulted in a substantial loss of activation abilities for both activators. Replacing the N-terminal sequence of activator G with the N-terminal sequence of activator E, and adding the C-terminal sequence of activator E to the C-terminus of activator G could increase the specific activity of L-NHase by 178.40%. The activation by nitrile hydratase activators was universal and specific, and the conserved domains of activators were critical for activation, while the N-terminal domain and C-terminal domain also had important effects on activation.

A new method for detection of Escherichia coli exist in licorice decoction was developed by using DNA-based electrochemical biosensor. The thiolated capture probe was immobilized on a gold electrode at first. Then the aptamer for Escherichia coli was combined with the capture probe by hybridization. Due to the stronger interaction between the aptamer and the E. coli, the aptamer can dissociate from the capture probe in the presence of E. coli in licorice decoction. The biotinylated detection probe was hybridized with the single-strand capture probe. As a result, the electrochemical response to Escherichia coli can be measured by using differential pulse voltammetric in the presence of α-naphthyl phosphate. The plot of peak current vs. the logarithm of concentration in the range from 2.7×10² to 2.7×10⁸ CFU·mL⁻¹ displayed a linear relationship with a detection limit of 50 CFU·mL⁻¹. The relative standard deviation of 3 successive scans was 2.5%，2.1%，4.6% for 2×10²，2×10⁴，2×106：⁶ CFU·mL⁻¹ E. coli, respectively. The proposed procedure showed better specificity to E. coli in comparison to Pseudomonas aeruginosa, Staphylococcus aureus and Bacillus subtilis. In the detection of the real extractum glycyrrhizae, the results between the proposed strategy and the GB assay showed high degree of agreement, demonstrating the designed biosensor could be utilized as a powerful tool for microbial examination for traditional Chinese medicine.

Objective : The purpose of this study was to investigate the relevant social and environmental factors affecting the occurrence of periodontal diseases during pregnancy in pregnant women and to analyze the influence of the periodontal status of women in the second trimester of pregnancy on small for gestational age (SGA) delivery. Methods: A total of 215 pregnant women were enrolled in this study in the Department of Periodontology of the West China Hospital of Stomatology of Sichuan University from May 2015 to May 2018. Periodontal parameters, such as bleeding on probing (BOP), probing depth (PD) and clinical attachment loss (CAL), were recorded at 16-24 weeks of gestational age. Subjects were divided into the periodontitis (n=32) group, gingivitis (n=171) group and periodontally healthy (n=12) group according to their periodontal conditions. With the patient′s informed consent, the patient decided whether to receive periodontal treatment. Basic and socioeconomic information was collected through questionnaires. After delivery, subjects were divided into the SGA group and non-SGA group according to their birth results. The periodontal clinical indicators, questionnaire results and delivery results were compared among the groups. Results : The mean PD (P=0.005, r=-0.192) and BOP% (P=0.003, r=-0.199) were negatively correlated with economic income. The family income in the periodontitis group was significantly lower than that in the healthy group and the gingivitis group (P < 0.05). The flossing use rate was significantly higher in the healthy group than that in the gingivitis group (P < 0.05). A total of 106 pregnant women received scaling and root planing, while 109 patients only received oral hygiene instruction. After delivery, SGA occurred in 23 cases (10.7%), and there were no significant difference in SGA incidence among the three groups (P > 0.05). PD ≥ 5 mm% and PD ≥ 4 mm% (P < 0.05) were significantly higher in the SGA group than in the non-SGA group. There was no significant difference in SGA incidence between the treated group and the untreated group (P > 0.05). Conclusion Family income and dental flossing use have an impact on the incidence of periodontal diseases during pregnancy. The severity of periodontitis in pregnant women is correlated with the incidence of SGA.