Condition optimization for immobilization of Fungal laccase with Nylon Net and glutaraldehyde and the nature of the immobilized enzyme were studied. The optimum conditions of the immobilization are: Nylon Net is crosslinked with 5% glutaraldehyde 15mL for 6 hours; the 30U laccase is added for immobilization for 8 hours. On this case, the recovery of enzyme activity was 50.3%. Compared with free enzyme, the thermal stability of immobilized enzyme was improved evidently but the optimal pH decreased slightly. 52% enzyme activity of immobilized laccase was hold after 8 cycles treatment with low concentration pulp wastewater.

To obtain high-yield avilamycin-producing strains,low energy N~+ ion implantation technology and screening of streptomycin-re- sistant mutants are used in the study on breeding mutation.The results show that,“saddle”region,which range is from 3?10~(15) to 5?10~(15) ions/cm~2,has got better induced mutation action.It also means that the strain's resistant mutation and yield mutation closely correlate to each other,and the method of streptomycin resistant screening is feasible.We have isolated a high-yield strain SVT-45 which the productivi- ty is 195% higher than the original strain's in the rotation-flask experiments.These results showed that the ion implantation was an effective method for microbe mutagensis.

As a depot drug delivery system, injectable polylactide-polyglycolide (PLGA) sustained-release microspheres have been successfully used to treat many diseases since the first microsphere product Lupron depot was approved for marketing in the United States in 1989. It has the ability of long-term release in the body for several days to several months, which can not only reduce the times of administration, but also reduce the drug blood concentration fluctuations, significantly improve the safety and patient compliance. In vitro-in vivo correlation (IVIVC) makes the development of microspheres more possible. It can describe the dynamic information of drug release in vivo through the in vitro release behavior of microspheres, and can reduce the workload of each stage and shorten the time span while characterizing the performance of microspheres. IVIVC can provide guidance or support for drug development, production changes, supervision and management. This article summarizes the release mechanism of injectable PLGA sustained-release microspheres, common measurement methods and theories of in vitro and in vivo release. And we also focus on the establishment and application of IVIVC, especially A level IVIVC in the field of microsphere preparations, to provide a reference for further study on in vitro-in vivo correlation of microspheres.

OBJECTIVETo observe the effect of American Ginseng Capsule (AGC) on the liver oxidative injury and the Nrf2 protein expression in the liver tissue of rats exposed by 900 MHz cell phone electromagnetic radiation.

METHODSTotally 40 male SD rats were randomly divided into the normal control group, the model group, the Shuifei Jibin Capsule (SJC) group, and the AGC group,10 in each group. Rats in the normal control group were not irradiated. Rats in the rest three groups were exposed by imitated 900 MHz cellular phone for 4 h in 12 consecutive days. Meanwhile, rats in the SJC group and the AGC group were intragastrically administrated with suspension of SJC and AGC (1 mL/200 g body weight) respectively. Normal saline was administered to rats in the normal control group and the model group. The histolomorphological changes of the liver tissue were observed by HE staining. Contents of malonic dialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH), and glutathione peroxidase (GSH-PX)were detected by colorimetry. The Nrf2 protein expression of hepatocytes was detected by immunohistochemical assay and Western blot.

RESULTSCompared with the normal control group, hepatocyte nucleus was atrophied or partially disappeared, the contents of liver MDA and Nrf2 protein obviously increased (P <0. 05, P <0. 01); contents of liver SOD and GSH decreased (P <0. 05) in the model group. Compared with the model group, karyopyknosis was obviously attenuated and approached to the normal level in the SJC group and the AGC group. The contents of liver MDA and Nrf2 protein expression decreased (P <0. 05), and the contents of liver SOD, GSH, and GSH-PX obviously increased (P < 0.05) in the SJC group. The contents of liver MDA and the Nrf2 protein expression decreased (P < 0.05), and contents of SOD and GSH obviously increased in the AGC group (P <0.01, P <0.05).

CONCLUSIONSThe electromagnetic radiation induced by 900 MHz cell phone could affect the expression of Nrf2 protein, induce oxidative injury, and induce abnormal morphology of liver cells. SJC and AGC could promote the morphological recovery of the liver cells. Its mechanism might be related to affecting the expression of Nrf2 protein and attenuating oxidative damage of liver cells.

Animals ; Cell Phone ; Electromagnetic Radiation ; Glutathione Peroxidase ; metabolism ; Hepatocytes ; metabolism ; Liver ; Male ; NF-E2-Related Factor 2 ; metabolism ; Oxidative Stress ; drug effects ; Panax ; Plant Extracts ; pharmacology ; Rats ; Superoxide Dismutase ; metabolism

Objective: To observe the effect of electroacupuncture (EA) at Huantiao (GB 30) and Weizhong (BL 40) on thigmesthesia, gait function, and expression levels of serum immunoglobulin G (IgG) and immunoglobulin M (IgM) in rabbits with lumbar intervertebral disc herniation (LIDH). Methods: Forty healthy New Zealand rabbits were randomly divided into a blank control group, a model group, an EA at acupoint group and an EA at non-acupoint group, with 10 rabbits in each group. The LIDH pathological model of rabbit was established using the self-made LIDH model maker. The thigmesthesia and gait function of rabbits were recorded by Siegal method. The serum IgG and IgM expression levels were detected by enzyme-linked immunosorbent assay. Results: EA at Huantiao (GB 30) and Weizhong (BL 40) could improve the clinical symptoms of thigmesthesia and gait function, and inhibit the expressions of serum IgG and IgM in the LIDH rabbits, which were significantly different compared with those in the model group and EA at non-acupoint group. Conclusion: EA at Huantiao (GB 30) and Weizhong (BL 40) can improve the clinical symptoms of LIDH rabbits, which is associated with inhibition of the serum IgG and IgM expressions and reduction of the immunoinflammatory factor release. This may be one of the mechanisms of EA at Huantiao (GB 30) and Weizhong (BL 40) in the treatment of LIDH.

OBJECTIVETo explore the mechanism of cake-separated moxibustion in treatment of hyperlipemia.

METHODSSixty rabbits were randomly divided into 4 groups, a blank group,a model group, a direct moxibustion group and a cake-separated moxibustion group. Hyperlipemia model was developed by high fat diet of cholesterol. Changes of ultrastructures of endothelial cells of the aorta of the rabbit were observed with electron microscope.

RESULTSThe endothelial cells in the cake-separated moxibustion group were more intact, most of them were normal in forms, internal elastic membrane was continuous, their thickness was even, the cells of smooth muscles in the medial membrane were relatively normal, which are similar to those in the blank control group. But the structure of endothelial cells of the aorta in the model group disappeared, in cytoplasm the sedimentation of a great number of lipids can be seen, internal elastic membrane was interrupted, the thickness was uneven, with focal dissolution, the cells of smooth muscle in the medial membrane had sedimentation of lipids, with frothy change.

CONCLUSIONCake-separated moxibustion has a certain protective action on endothelial cells of the aorta in the rabbit of hyperlipemia.

Animals ; Aorta ; Endothelial Cells ; Hyperlipidemias ; Lipids ; Moxibustion ; Rabbits

Objective:To observe the effect of moxibustion on the mRNA and protein expressions of heat-shock protein 70 (HSP70) in gastric cancer-bearing rats. Methods: A total of 40 healthy Sprague-Dawley (SD) rats were adaptively fed for one week. The gastric cancer model was prepared by Walker-256 cancer tissue transplantation. After 7 d, 10 rats were randomly selected to verify the successful modeling, and the remaining 30 rats were divided into a model group, a moxibustion group and an infrared group by the random number table method, with 10 rats in each group. After enrollment, the moxibustion group received suspended moxibustion at Zhongwan (CV 12), Guanyuan (CV 4) and bilateral Zusanli (ST 36), (the first group of acupoints) on the 1st day, and suspended moxibustion at bilateral Pishu (BL 20) and Weishu (BL 21), (the second group of acupoints) on the 2nd day, 20 min each time, once a day. Moxibustion was alternately performed every other day at the two groups of acupoints for 21 d. From the day of enrollment, rats in the infrared group were irradiated with the infrared radiation at the stomach area on the 1st day, and at the T12-T13 interspinous region on the 2nd day, 20 min each time, once a day, and the two locations were alternately irradiated every other day for 21 d. During the treatment, rats in the model group were intervened by grasping and fixation without treatment. At the end of the treatment, blood was collected from the inner eye orbit, and the HSP70 expression in peripheral blood was determined by enzyme linked immunosorbent assay (ELISA). Rats were sacrificed, the tumor volume and growth inhibition rate were measured. The position and changes of HSP70 in gastric cancer were observed by streptavidin-perosidase (SP); HSP70 protein expression was determined by ELISA; HSP70 mRNA expression in cancer tissues was determined by reverse transcription-polymerase chain reaction (RT-PCR) assay. Results: In comparison of the model group, the volume growth of the gastric cancer in the moxibustion group was significantly restricted (P＜0.01); the volume growth inhibition rate in the moxibustion group was 37.93%; the HSP70 expression in peripheral blood and the cancer tissues was significantly increased (both P＜0.01); the expression of HSP70 mRNA and HSP70 content in gastric tumor were both obviously increased in the moxibustion group (P＜0.01); and a large amount of HSP70 was released to the outside of cancer cells in the moxibustion group. In comparison of the model group, the volume growth of the gastric cancer in the infrared group was slightly restricted (P＜0.05) with a volume growth inhibition rate of 15.89%; the HSP70 expression in the infrared group was increased significantly in peripheral blood (P＜0.01) and in the gastric cancer tissues (P＜0.05); more HSP70 was released outside of the cancer cells in the infrared group. In comparison of the infrared group, the volume growth of gastric cancer was more restricted in the moxibustion group (P＜0.05), and the HSP70 expression in the gastric cancer tissues was also higher (P＜0.05); more HSP70 was released outside of the cancer cells in the moxibustion group. Conclusion: Moxibustion and infrared treatment inhibit the gastric cancer growth in the gastric cancer-bearing rats, up-regulate the HSP70 expression in gastric cancer tissues, and promote the production and extracellular release of HSP70, and the effect of moxibustion is more obvious.

Objective:To observe the effect of warm-unblocking acupuncture plus fluticasone propionate nasal spray on the pulmonary ventilation, level of interferon-γ (IFN-γ) and sleep quality in patients with allergic rhinitis (AR). Methods: A total of 112 AR patients were enrolled between January 2013 and August 2018 and were divided into an observation group and a control group by the random number table method, with 56 cases in each group. Patients in the observation group received warm-unblocking acupuncture plus fluticasone propionate nasal spray, and patients in the control group only received fluticasone propionate nasal spray. The nasal symptom score, pulmonary function indexes, the levels of IFN-γ and interleukin (IL)-4 in serum, and sleep quality in the two groups were compared. Results: After treatment, the total effective rate in the observation group was higher than that in the control group (P＜0.05). The nasal symptom score dropped in both groups after treatment (both P＜0.05), and the score in the observation group was lower than that in the control group (P＜0.05). The pulmonary ventilation indexes all increased significantly after treatment in the observation group (all P＜0.05); the forced expiratory volume in 1 second (FEV1) to forced vital capacity (FVC) ratio (FEV1/FVC) and the forced expiratory flow at 50%, 75% and 25%-75% of the vital capacity (FEF50%, FEF75%, FEF25%-75%) increased after treatment in the control group (all P＜0.05); the pulmonary ventilation indexes were higher in the observation group than those in the control group (all P＜0.05). The level of IFN-γ increased significantly after treatment in the two groups (both P＜0.05) and the level of IL-4 dropped significantly (both P＜0.05); the observation group had a higher IFN-γ level (P＜0.05) and a lower IL-4 level (P＜0.05) compared with the control group. Regarding the Pittsburgh sleep quality index (PSQI), the scores of subjective sleep quality, habitual sleep efficiency and sleep disturbances and the general PSQI score decreased significantly after treatment in both groups (all P＜0.05), and the scores in the observation group were significantly lower than those in the control group (all P＜0.05). Conclusion: Warm-unblocking acupuncture plus fluticasone propionate nasal spray can effectively control the clinical symptoms and improve pulmonary function in the treatment of AR; this approach can regulate the levels of IFN-γ and IL-4 towards the normal range in AR patients; it can also improve patient’s sleep quality. This method can produce more significant efficacy than fluticasone propionate nasal spray used alone.

As one of the most serious types of psoriasis, pathogenesis of erythrodermic psoriasis (EP) is unclear so far. In this study, we aimed to detect the levels of Th1/Th2 cytokine-associated transcription factors and T-lymphocyte clone in peripheral blood mononuclear cells (PBMCs) derived from EP patients, and gene expression level of T-bet/GATA-3 in skin lesion. The potential role of Th1/Th2 reaction pattern played in the pathogenesis of EP was also discussed. Serum levels of IFN-γ, IL-2, IL-4 and IL-10 were quantified by ELISA among 16 EP patients, 20 psoriasis vulgaris (PV) patients and 15 healthy controls. The expression levels of T-bet/GATA-3 in the skin lesion and PBMCs were examined by real-time qPCR. The ratio of Th1/Th2 was measured by flow cytometry. The levels of IFN-γ, IL-2, IL-4 and IL-10 were higher in EP patients than in the healthy controls. The levels of IL-4 and IL-10 were 69.44±11.45 and 12.62±4.57 pg/mL, respectively, in EP patients, significantly higher than those in PV patients and healthy controls (P<0.05). Flow cytometry revealed the levels of both Th1 and Th2 in PBMCs from EP patients were higher than those in healthy controls, and the Th1/Th2 ratio was dramatically lower than in PV patients (P<0.01). The ratios of IFN-γ/IL-4 and T-bet/GATA-3 in EP patients were both less than 1.0, suggesting a reversal when compared with the other two groups. Our study indicated that the EP patients exerted a Th1/Th2 bidirectional response pattern, and the balance of Th cell subsets inclines to Th2, which might be one of the important mechanisms of EP pathogenesis.

OBJECTIVETo establish a rapid quantitative analysis method for the content of chlorogenic acid and solid content in the extraction liquid concentration process during the production of Reduning injection by using the near-infrared (NIR) spectroscopy, in order to reflect the concentration state in a real-time manner and really realize the quality control of concentrating process of the extraction and concentration process.

METHODThe samples during the Jinqing extraction liquid concentration process were collected. After the removal of abnormal samples, the spectra pretreatment and the wave band selection, the quantitative calibration model between NIR spectra and chlorogenic acid HPLC analytical value and solid content was established by using PLS algorithm, and unknown samples were predicted.

RESULTThe correlation coefficients between the chlorogenic acid content and the solid content were respectively 0.992 1 and 0.994 0, and the correlation coefficients of the verification model were respectively 0.994 4 and 0.998 4, with the root mean square error of calibration (RMSEC) of 0.814 6 and 2.656 1 and the root mean square error of prediction (RMSEP) of 0.704 6 and 1.876 7 respectively, and the relative standard errors of predictions (RSEP) were 6.01% and 2.93% respectively.

CONCLUSIONThe method is simple, rapid, nondestructive, accurate and reliable, thus could be adopted for the fast monitoring of the chlorogenic acid content and the solid content during the concentration process of Reduning injection extraction liquid.

Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Quality Control ; Spectroscopy, Near-Infrared ; methods