Construction of cell factories for production of patchoulol in Saccharomyces cerevisiae / 中国中药杂志

Patchoulol is an important sesquiterpenoid in the volatile oil of Pogostemon cablin, and is also considered to be the main contributing component to the pharmacological efficacy and fragrance of P. cablin oil, which has antibacterial, antitumor, antioxidant, and other biological activities. Currently, patchoulol and its essential oil blends are in high demand worldwide, but the traditional plant extraction method has many problems such as wasting land and polluting the environment. Therefore, there is an urgent need for a new method to produce patchoulol efficiently and at low cost. To broaden the production method of patchouli and achieve the heterologous production of patchoulol in Saccharomyces cerevisiae, the patchoulol synthase(PS) gene from P. cablin was codon optimized and placed under the inducible strong promoter GAL1 to transfer into the yeast platform strain YTT-T5, thereby obtaining strain PS00 with the production of(4.0±0.3) mg·L~(-1) patchoulol. To improve the conversion rate, this study used protein fusion method to fuse SmFPS gene from Salvia miltiorrhiza with PS gene, leading to increase the yield of patchoulol to(100.9±7.4) mg·L~(-1) by 25-folds. By further optimizing the copy number of the fusion gene, the yield of patchoulol was increased by 90% to(191.1±32.7) mg·L~(-1). By optimizing the fermentation process, the strain was able to achieve a patchouli yield of 2.1 g·L~(-1) in a high-density fermentation system, which was the highest yield so far. This study provides an important basis for the green production of patchoulol.

Construction of cell factories for production of valencene in Saccharomyces cerevisiae / 药学学报

Valencene, a kind of sesquiterpenoid with a citrus flavor, is mainly found in Valencia orange and is commonly used in cosmetics and food additives, as well as industrial synthetic nootkatone. In this study, synthetic biology was used to create a Saccharomyces cerevisiae cell factory to produce valencene. Fistly, valencene synthase gene (CnVS) from Callitropsis nootkatensis was inserted into the chromosome of the chassis strain YTT-T5. The resulting strain VAL-01 could produce 1.1 mg·L-1 valencene. Protein fusion technique was used, different valencene synthases were compared and the copy number of key genes was adjusted, yielding valencene to 436.4 mg·L-1. Then, knocking-out the transcription factor ROX1 resulted in valencene improvement by 17.4%. Moreover, the induction system of galactose was regulated, transcription factor PDR3 and INO2 were overexpressed. The engineered strain VAL-10 could produce 2 798.6 mg·L-1 valencene by high cell density fermentation method (nearly 2 500 times higher than VAL-01). This study provides a basis for green production of valencene.

Clinical Study on Modified Guipitang Combined with Xuefu Zhuyutang in Treating Mild Cognitive Impairment After Cerebral Infarction / 中国实验方剂学杂志

ObjectiveTo observe the clinical efficacy and anti-inflammatory and anti-oxidant effect of modified Guipitang combined with Xuefu Zhuyutang in the treatment of mild cognitive impairment （MCI） after cerebral infarction with syndromes of heart and spleen deficiency and blood stasis blocking collateral. MethodA total of 114 eligible patients were randomly divided into a control group and an observation group，with 57 cases in each group. Patients in the control group were given red deer ginseng tablets （po），4 tablets/time，2 times/day. Patients in the observation group were given modified Guipitang combined with Xuefu Zhuyutang （po，1 dose/day）for continuous 8 weeks. This study compared the scores of montreal cognitive assessment （MoCA） scale，Rivermead behavioral memory test （RBMT），activities of daily living （ADL），trail making test B （TMT-B），neuropsychiatric inventory questionnaire （NPI） and scores of traditional Chinese medcine（TCM） syndrome with syndromes of heart and spleen deficiency and blood stasis blocking collateral before and after treatment. Then we further detected the levels of 8-hydroxydeoxyguanosine （8-OHDG），malondialdehyde （MDA），oxidized low density lipoprotein （ox-LDL），superoxide dismutase （SOD），homocysteine （Hcy），interleukin-8 （IL-8），C-reactive protein （CRP） and fibrinogen （FIB） levels before and after treatment. ResultThe total effective rate for the treatment of cognitive function impairment in the observation group was 92.98% （53/57），which was higher than 78.95% （45/57） in the control group （χ2＝4.653，P<0.05）. The recovery rate of cognitive function in the observation group was 54.39% （31/57），which was higher than 33.33% （19/57） in the control group （χ2＝5.130，P<0.05）. The MoCA，RBMT and ADL scores of the observation group were higher than those of the control group （P<0.01），and the TMT-B time of the former was shorter than that of the latter （P<0.01）. In addition， the observation group showed lower scores of TCM syndrome，NPI-1 and NPI-2 scores than the control group （P<0.01）. The SOD level of the observation group was higher than that of the control group （P<0.01），and the levels of 8-OHDG，ox-LDL，MDA，Hcy，IL-8，CRP and FIB were lower than those of the control group （P<0.01）. ConclusionModified Guipitang combined with Xuefu Zhuyutang can improve cognitive function in MCI patients after cerebral infarction with syndromes of heart and spleen deficiency and blood stasis blocking collateral， with anti-inflammatory and anti-oxidant effect， and yield superior efficacy than red deer ginseng tablets.

Paeonol suppresses fibroblast synovitis by increasing MANF expression / 中国药理学通报

Aim To investigate the possible mechanism of paeonol inhibiting the inflammatory response of fibroblast synovial cells (RA-FLSS) in rheumatoid arthritis. Methods CCK-8 assay was used to detect Paeonol's inhibitory level on the abnormal proliferation of arthritis human fibroblast synovial cells (RA-FLSs). The levels of endoplasmic reticulum stress-related proteins MANF and ATF6 were detected by Western blot. Cell localization of transcription factor p65 and Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) was detected by immunofluorescence. RT-qPCR detected the changes of p65 target genes. Results Paeonol could significantly inhibit the abnormal proliferation of RA-FLSS cells. Paeonol activates ATF6 and increases the expression of MANF. Paeonol promoted the nuclear transfer of MANF protein and inhibited the transcriptional activity of p65. Conclusion Paeonol promotes the expression of MANF and nuclear transfer through the endoplasmic reticulum stress pathway and affects the progression of RA by inhibiting the transcriptional activity of p65.

China′s strategy and experience in preventing and controlling the local spreading and epidemic of COVID-19 / 上海预防医学

After implementing a series of prevention and control strategies of Corona Virus Disease 19(COVID-19), China′s local epidemic situation has been basically blocked.While China has achieved a periodic success, global pandemic situation is still serious.Together with the timeline of China′s epidemic prevention and control this study reviewed the main strategies and measures in response to COVID-19 epidemic from Dec.2019 to Mar.23, 2020, and summarized China′s prevention and control plans and experiences.We hope all countries could fully understand the importance of non-pharmaceutical public health interventions, make positive and effective adjustments to the measures and strategies of prevention and control based on their own national conditions.

Safety depth of acupuncture at Yamen (GV 15) in patients with atlantoaxial dislocation / 中国针灸

OBJECTIVE@#To explore and compare the safety depths of perpendicular and oblique acupuncture at Yamen (GV 15) in patients with atlantoaxial dislocation (AAD) and healthy volunteers.@*METHODS@#One bundred and seventy-seven patients with atlantoaxial dislocation were selected as an AAD group, and 207 patients without atlantoaxial dislocation and with normal anatomical structure were selected as a normal group. All participants were moderately sized. The MRI scanning of the cervical vertebra was performed, and the safety depth of perpendicular and oblique acupuncture at Yamen (GV 15) was calculated on the sagittal image.@*RESULTS@#In the AAD group, the safety depth of men was (45.33±5.17) mm for perpendicular acupuncture and (48.58±4.41) mm for oblique acupuncture; the safety depth of women was (44.17±7.80) mm for perpendicular acupuncture and (47.49±7.32) mm for oblique acupuncture. In the normal group, the safety depth of men was (47.72±5.06) mm for perpendicular acupuncture and (42.69±5.53) mm for oblique acupuncture; the safety depth of women was (44.63±5.85) mm for perpendicular acupuncture and (39.88±6.18) mm for oblique acupuncture. The safety depth of men and women for oblique acupuncture was longer than that for perpendicular acupuncture in the AAD group (0.05); the safety depth of perpendicular and oblique acupuncture for men was longer than that for women in the normal group (0.05), while the safety depth of oblique acupuncture in the AAD group was longer than that in the normal group (<0.01).@*CONCLUSION@#The safe depth of acupuncture at Yamen (GV 15) has significantly changed under AAD, so during the clinical acupuncture the needle insertion should be less than its safe depth.

Study on the angle of needling Yamen (GV 15) in atlanto-axial dislocation patients / 针灸推拿医学（英文版）

Objective: To compare the risk angle and safety angle of needling Yamen (GV 15) between the atlanto-axial dislocation (AAD) patients and healthy subjects. Methods: A total of 177 AAD patients diagnosed and treated at the Center of Upper Cervical Vertebra of Beijing China-Japan Friendship Hospital between January 2010 and January 2018 were included in the AAD group. Another 207 healthy subjects were included in the normal group. There were totally 191 males and 193 females. The MRI scan was performed for the cervical vertebrae to measure the risk angle and safety angle of acupuncture at Yamen (GV 15) on the sagittal image. Results: In the AAD group, the risk angle was (13.14±3.99)° and the relative safety angle was (10.31±3.23)° for the perpendicular needling, while the oblique needling risk angle was (9.09±3.09)° for the male; the risk angle was (12.12±2.74)° and the relative safety angle was (10.56±2.09)° for the perpendicular needling, while the oblique needling risk angle was (9.70±2.95)° for the female. In the normal group, the risk angle was (7.89±1.59)° and the relative safety angle was (10.21±3.55)° for the perpendicular needling, while the oblique needling risk angle was (16.07±1.77)° for the male; the risk angle was (6.93±1.45)° and the relative safety angle was (10.70±2.94)° for the perpendicular needling, while the oblique needling risk angle was (14.89±2.18)° for the female. The perpendicular needling risk angles for the males and females in the AAD group were larger than those in the normal group, and the differences were statistically significant (bothP<0.01); for the inner-group comparison, there was no significant difference in the perpendicular needling risk angle between the male and the female in the AAD group (P>0.05); however, the perpendicular needling risk angle for the male was larger than the female, and the difference was statistically significant in the normal group (P<0.01). There were no significant differences in the relative safety angle for both the male and the female between the AAD group and the normal group (bothP>0.05). For the inner-group comparison, there was no significant difference in the relative safety angle between the male and the female (P>0.05). The oblique needling risk angles for both the males and females were smaller in the AAD group than those in the normal group, and the differences were statistically significant (bothP<0.01); the oblique needling risk angle for the male was not significantly different from that for the female in the AAD group (P>0.05); in the normal group, the oblique needling risk angle for the male was larger than that for the female, and the difference was statistically significant (P<0.01). Conclusion: Under the AAD condition, the risk angle and safety angle of acupuncture at Yamen (GV 15) change significantly, perpendicular needling should be better if performed slightly lower than the horizontal direction, and the oblique needling should be safer across the occipital foramen toward the occipital bone.

Discussion on the application principle of tuina manipulations for lumbar intervertebral disc herniation in Chinese literatures in recent 30 years / 针灸推拿医学（英文版）

Objective: To discuss the application principle in tuina manipulation for lumbar intervertebral disc herniation (LIDH) in Chinese literatures published in recent 30 years. Methods: The three major Chinese databases, Wanfang Academic Journal Full-text Database (Wanfang), Chongqing VIP Database (CQVIP) and China National Knowledge Infrastructure (CNKI), were searched to collect the studies of tuina manipulations in treatment of LIDH published in recent 30 years. Clustering analysis was applied to analyze the top 20 tuina manipulations for LIDH. Results: The top 20 most frequently used manipulations for LIDH were Gun-rolling, Rou-kneading, Dian-digital pressing, oblique Ban-pulling, An-pressing, Tanbo-plucking, Bashen-pulling and extending, horizontal Tui-pushing, Na-grasping, Anrou-pressing and kneading, Dou-shaking, Yao-rocking, Ca-scrubbing, Pai-patting, post-extension Ban-pulling, Mo-rubbing, Zhen-vibrating, Nie-pinching, fist-back Ji-tapping, and dorsal Shen-extending methods. The involved manipulations can be divided into two categories by the treated body areas. One category is applied to the soft tissues, including Gun-rolling, Rou-kneading, Dian-digital pressing, An-pressing, Tanbo-plucking, horizontal Tui-pushing, Na-grasping, Anrou-pressing and kneading, Ca-scrubbing, Pai-patting, Mo-rubbing, Zhen-vibrating, Nie-pinching, and fist-back Ji-tapping methods. The other category is applied to bones and joints, including oblique Ban-pulling, Bashen-pulling and extending, Dou-shaking, Yao-rocking, post-extension Ban-pulling, and dorsal Shen-extending methods. Conclusion: Based on the treated body area, the tuina manipulations applied to treat LIDH are predominated by the ones performed on soft tissues, assisted by those on bones and joints. From the way of force exertion, the involved manipulations are majorly the swinging methods, followed by squeezing and pressing ones. The manipulations applied to bones and joints are predominated by the Ban-pulling ones, followed by the Bashen-pulling and extending ones.

Metal/Matrix Enhanced Time-of-Flight Secondary Ion Mass Spectrometry for Single Cell Lipids Analysis / 分析化学

The chemical components analysis of single cell is important for understanding of physiological processes such as cell growth, signal transduction and apoptosis.Time-of-flight secondary ion mass spectrometry ( ToF-SIMS) is a sensitive surface analysis technique with high spatial resolution and can be used for single cell and micro-area analysis.However, relatively low ioniZation yield of biomolecules limited its wide application in single cell analysis.Herein, we used metal substrate and matrix material to enhance the ioniZation yield of lipids.The signal intensity of the phosphatidylcholine PC (40:0) casted on the matrix/gold coated silicon substrate was 65 times higher than that on the silicon wafer.Signal enhancement of phosphatidylcholine PC (34:1) on the single cell surface cultured on matrix/gold coated silicon substrate was observed as well.Due to the influence of irregular topography and complex chemical environment of cell, the increase of lipids signal was smaller.Delayed extraction mode of ToF-SIMS overcame the effects of cell topography, leading to further enhancement of the signal intensity of lipids.Meanwhile, simultaneous high spatial resolution of chemical imaging and high mass resolution of the mass spectra of single cells were obtained.Our strategies provided new insights into the study of cell metabolism and cell-environment interactions.

Study on the mobile phone dependence syndrome and its distribution among 2213 college students in Guangzhou / 中华流行病学杂志

Objective To investigate the occurrence,distribution and risk factors of mobile phone dependence syndrome (MPDS) among college students in Guangzhou.Methods A unified questionnaire was adopted,with 2311 college students from 6 universities in Guangzhou investigated by cluster sampling.Distribution and risk factors of MPDS among different groups were analyzed by logistic regression.Results A total number of 2213 effective questionnaires was retrieved,including 1149 males and 1064 females.The average age was (21.33 ± 1.72).The incidence rate of MPDS among studied college students in Guangzhou was 23.3％ (515/2213).Regarding the distribution of personal characteristics,significant differences were found in the following aspects:grades,majors in college,being the only child of the family,monthly cost of living,personal characters and the academic performance at school (P＜0.05).Regarding the distribution of characteristics among parents,significant differences were found in the following areas:educational levels of the mother,rearing patterns of both parents,status of feeling on mother's caring (P＜0.05) etc.The main risk factors for MPDS were as follows:students majored in literature and law,with high monthly living cost,father' s autocratic and democratic patterns of rearing,mother' s autocratic and doting rearing pattern as well as personal feeling on mother's attitude of unconcern.The incidence of MPDS among those persons with uncertain characters was less than those who were extroverts.Conclusion MPDS among college students seemed to be severe in Guangzhou.No difference was found in the incidence rates of MPDS between genders.Should take interventions according to its risk factors.

Research progress of microparticles as drug delivery system for traditional Chinese medicine / 中国中药杂志

Microparticle preparation, a new drug delivery system based on microencapsulation technique, includes micro-spheres and microcapsules. Recently, this new drug delivery system has been applied in developing new dosage forms for the traditional Chinese medicine (TCM). TCM microparticles can perform several sound characteristics and functions which are currently unavailable in TCM preparations, such as controlled release, effect of targeting, increasing bioavailability or low toxicity. This makes it possible that TCM may exert much higher therapeutic efficacy and show lower side-effects as well. Although the studies on TCM microparticles are still in the beginning stage, microparticle preparation of TCM has given rise to comprehensive attention and will have a wonderful prospect. The progress in this field is reviewed in this article.