During the production of ε-poly-L-lysine (ε-PL) in fed-batch fermentation, the decline of ε-PL synthesis often occurs at middle or late phase of the fermentation. To solve the problem, we adopted two strategies, namely pH shift and feeding yeast extract, to improve the productivity of ε-PL. ε-PL productivity in fermentation by pH shift and feeding yeast extract achieved 4.62 g/(L x d) and 5.16 g/(L x d), which were increased by 27.3% and 42.2% compared with the control ε-PL fed-batch fermentation, respectively. Meanwhile, ε-PL production enhanced 36.95 g/L and 41.32 g/L in 192 h with these two strategies, increased by 27.4% and 42.48% compared to the control, respectively. ε-PL production could be improved at middle or late phase of fed-batch fermentation by pH shift or feeding yeast extract.
Batch Cell Culture Techniques ; Fermentation ; Industrial Microbiology ; Nitrogen ; chemistry ; Polylysine ; biosynthesis

This study investigated the protection against the ND in chickens by a recombinant DNA vaccine. A plasmid vector encoding NDV F protein, which is reqired for virus cell fusion and is important for vaccine induced immunity, was used as a model to study how DNA vaccines may be modulated by the simulaneous expression of chicken IL-2. The NDV D26 strain F gene with CMV promotor and BGH polyA signal sequence was amplified by PCR from eukaryotic plasmid pcDNA-F, which contains the full-length NDV F gene, and clond into reconstructed eukaryotic plasmid pcDNA-IL2, which contains chicken IL-2 gene. Restriction endonuclease cleavage and PCR amplification showed that a bicistronic plasmid encoding NDV F gene and chicken IL-2 separately was successfully constructed. Two-week-old SPF chickens were intramuscularly innoculated the recombinant plasmid. Antibody and lymphocyte proliferative assay showed that the humoral and cellular immunity of chickens vaccinated the recombinant plasmid greatly increased compared with those innoculated only plasmid expressing NDV F protein. Challenged with the lethal dose of NDV F48E9 strain, 72% chickens vaccinated recombinant plasmid were survived, and 30% chickens vaccinated plasmid expressing F protein were survived. These results proved the adjuvant effect of chicken IL-2, and further showed that the efficacy of a DNA vaccine can be greatly improved by simultaneous expression of IL-2.

Objective To observe the mid-term effectiveness of percutaneous lumbar puncture and injection of medical ozone and collagenase in treating lumbar disc herniation.Methods CT-guided percutaneous lumbar puncture and medical ozone injection into the intervertebral disc was performed in 78 patients with lumbar disc herniation, which was confirmed by imaging study or clinical manifestations.After 2-4 days, neucleolysis was carried out.Collagenase 12 000 U was injected via the anterior epidural space onto the surface or into the inside of the prominence of the protruded disc.Results The clinical results were evaluated in 6-25 months after the procedure.Of 78 patients treated with this technique, the result was excellent in 58(74.3%), good in 16(20.5%) and poor in 4(5.1%).Symptoms returned in 12 cases in 2 months after the procedure, and the reappeared symptoms were relieved in 9 of them within 2 months without giving any treatment.In 3 cases the symptoms disappeared after repeated injection of medical ozone.No serious complications occurred.Conclusion Percutaneous lumbar puncture and injection of medical ozone combined with collagenase is an effective, safe and minimally-invasive therapy for lumbar disc herniation.

Objective To observe the treatment effect of joint manipulation under brachial plexus anesthesia combined with comprehensive physical therapy in treatment of periarthritis of shoulder. Mehods From 2010.3 to 2010.9, 60 patients with periarthritis of shoulder in the Department of Pain, The First People's Hospital of Jingzhou,were divided into 3 groups:group A,B and C.Twenty patients in Group A were given Joint manipulation under brachial plexus anesthesia for treatment of the periarthritis of shoulder. Twenty patients in Group B were given comprehensive physical therapy fro treatment of periarthritis of shoulder. Twenty patients in Group C were given Joint manipulation under brachial plexus anesthesia combined with comprehensive physical therapy for treatment of periarthritis of shoulder.Results All 60 patients were followed up for 1 months. At the 1st month after surgery, vas score was evaluated for patients. Postoperative and preoperative vas score difference was compared between 3 groups by t test. There was statistically significant difference in the postoperative and preoperative vas score between Group C and Group A,and between Group B and Group A (P<0.05) . Conclusion The treatment effect of Joint manipulation under brachial plexus anesthesia combined with physical therapy in treatment of periarthritis of shoulder is good, but has no obvious difference compared with comprehensive physical therapy.

Objective To observe the treatment effect of radio frequency target combined with collagenase in treatment of cervical intervertebral disc herniation. Methods From March 2011 to September 2011, 40 patients with cervical intervertebral disc herniation were divided into 2 groups: Group A and Group B. Patients in Group A were given radio frequency target radiofrequency for treatment of the cervical intervertebral disc herniation and patients in Group B of 20 were given radio frequency target combined with collagenase for treatment of cervical intervertebral disc herniation. Results 40 patients were followed up for 6 months. At the 1st week and 6 months after surgery, the VAS score difference between before and after surgery in patients were compared between two groups by t test. There was no statistically significant difference at the 1st week after surgery between two groups (P>0.05), while 6 months after surgery, there was statistically significant difference between two groups (P<0.05) .Conclusion Radio frequency target combined with collagenase for treatment of cervical intervertebral disc herniation can alleviate the pain symptoms of patients,and the short-term and long-term curative effect is very good. The long-term curative effect has statistically significant difference with pure target radiofrequency ablation treatment. Radio frequency target combined with collagenase for treatment of cervical intervertebral disc herniation has good safety, few side effects, low risks and good short-term and long-term curative effect, so it deserves to be generalized.

In this paper, chloramphenicol was selected as a model drug to prepare in situ gels. The intrinsic dissolution rate of chloramphenicol from in situ gel was evaluated using the surface dissolution imaging system. The results indicated that intrinsic dissolution rate of chloramphenicol thermosensitive in situ gel decreased significantly when the poloxamer concentration increased. The addition of the thickener reduced the intrinsic dissolution rate of chloramphenicol thermosensitive gel, wherein carbomer had the most impact. Different dilution ratios of simulated tear fluid greatly affected gel temperature, and had little influence on the intrinsic dissolution rate of chloramphenicol from the thermosensitive in situ gel. The pH of simulated tear fluid had little influence on the intrinsic dissolution rate of chloramphenicol thermosensitive in situ gel. For the pH sensitive in situ gel, the dissolution rates of chloramphenicol in weak acidic and neutral simulated tear fluids were slower than that in weak alkaline simulated tear fluid. In conclusion, the intrinsic dissolution of chloramphenicol from in situ gel was dependent on formulation and physiological factors. With advantages of small volume sample required and rapid detection, the UV imaging method can be an efficient tool for the evaluation of drug release characteristics of ophthalmic in situ gel.

OBJECTIVE:To conduct screening for the active ingredients of Ziziphus jujuba with neuroprotective effect and to il-luminate their mechanisms of action preliminarily. METHODS:After neuron-like cells (PC12 cells) were respectively cultured in the ingredient of Z. jujuba with polysaccharide enriched(1 mg/ml),that with polysaccharide removed(1 mg/ml),7 kinds of flavo-noid ingredients of Z. jujuba(catechin,procyanidine B2,epicatechin,hyperoside,rutin,quercetin-3-β-D-glucoside and kaempfer-ol-3-O-rutinoside,represented by A,B,C,D,E,F,G,all at the concentrations of 3,13,30 μmol/L)and 2 kinds of nucleoside ingredients of Z. jujuba (cyclic adenosine monophosphate and cyclic guanosine monophosphate,both at the concentrations of 3, 13,30 μmol/L)for 24 h,tert-butyl hydroperoxide(tBHP,150 μmol/L)was used to act on PC12 cells for 3 h to induce oxidative cellular damage,and MTT assay was employed to detect the survival rate of PC12 cells. The PC12 cells transfected with reporter gene plasmid(pARE-Luc)were cultured as above for 24 h,luciferase(Luc)assay was used to detect the transcriptional levels of the antioxidant response element(ARE)of all groups of cells(reflected as the activity of Luc)so as to investigate the anti-injury mechanism. RESULTS:The ingredient of Z. jujuba with polysaccharide enriched could significantly increase the survival rate of PC12 cells to which oxidative damage was caused and the transcriptional level of ARE in the transfected cells. Among the flavonoid ingredients of Z. jujuba, A(30 μmol/L),B(3-30 μmol/L)and C(10-30 μmol/L)could significantly increased the survival rate of the cells,and A(30 μmol/L),B(3-30 μmol/L),C(30 μmol/L),E(30 μmol/L)and F(3-30 μmol/L)could obviously in-creased the activation level of ARE in the transfected cells. However,the nucleoside ingredients of Z. jujuba including cyclic ade-nosine monophosphate and cyclic guanosine monophosphate had no obvious effect of increasing the survival rate of PC12 cells to which oxidative damage was caused or activating the transcription of ARE in the transfected cells. CONCLUSIONS:The polysac-charide and flavonoid ingredients of Z. jujuba may be the active ingredients which account for the neuroprotective effect against oxi-dative cellular damage,and their mechanisms of action may be related to the activation of ARE transcription.

Objective: To analyze the clinical application of the open supraclavicular approach in thyroidectomy. Methods: The clinical practicability of open supraclavicular thyroidectomy was explored by comparing the traditional anterior low arc incision thyroidectomy procedure with open supraclavicular thyroidectomy in terms of patients'aesthetic satisfaction, effectiveness of the operation, operation time, and so on. Result: Twenty-two cases of open supraclavicular thyroidectomy (group B) had better aesthetic satisfaction than 29 cases of traditional incision thyroidectomy (group A)(P<0.05), and had the same operative effect with traditional incision. Open supraclavicular thyroidectomy is associated with good aesthetic satisfaction, and has the same effect as the traditional incision does. Conclusions: Open supraclavicular thyroidectomy has good clinical value for benign thyroid tumors and some malignant tumors that require unilateral lobectomy, and even for tumors larger than the incision diameter. It has good aesthetic value while ensuring the curative effect of surgery.

Pure drug-assembled nanomedicines (PDANs) are currently under intensive investigation as promising nanoplatforms for cancer therapy. However, poor colloidal stability and less tumor-homing ability remain critical unresolved problems that impede their clinical translation. Herein, we report a core-matched nanoassembly of pyropheophorbide a (PPa) for photodynamic therapy (PDT). Pure PPa molecules are found to self-assemble into nanoparticles (NPs), and an amphiphilic PEG polymer (PPa-PEG