1.Effects of rehabilitation training combined with bone marrow mesenchymai stem cell-derived neural stem cells transplantation on the expression of Nogo-A and NgR proteins after spinal cord injury
Zhilai ZHOU ; Anmin JIN ; Yinhai CHEN ; Shaoxiong MIN
Chinese Journal of Physical Medicine and Rehabilitation 2011;33(8):566-570
Objective To study the effects of rehabilitation training combined with the transplantation of bone marrow mesenchymal stem cells-derived neural stem cells (BMSC-D-NSCs) on the expression of Nogo-A and NgR protein in rats after spinal cord injury (SCI).Methods The spinal cords of eighty Sprague-Dawley rats were injured using a modified Allen′s impactor (H = 25 mm) at T10. The injured rats were randomly divided into a combination therapy group which was given rehabilitation training and cell transplants, a cell graft group, a rehabilitation training group and a control group. At the 7th day post SCI, BMSC-D-NSCs were transplanted into the injured spinal cords of the rats in the combination therapy and cell graft groups. Hindlimb movement was assessed using the BassoBeattie-Bresnahan (BBB) scale every week, and protein was extracted from the injured spinal cord tissue for Nogo-A and NgR determination by Western blotting at the 1st, 3rd and 7th day after cell transplantation.Results The average BBB score of the rats in the combination therapy group was significantly higher than that of the other groups from 2 weeks post transplantation. The scores in the rehabilitation training group were significantly higher than in the control group from the 5th week post transplantation. Western blotting showed high expression of Nogo-A and NgR protein 24 h post surgery, but these declined with time. For Nogo-A there was a significant difference among the groups at all three time points. In the combination therapy group the expression declined to a minimum by the 7th day. For NgR protein there was no significant difference between the 1st and 3rd day in any group.Conclusions Rehabilitation training combined with BMSC-D-NSC transplantation can have a synergistic effect on functional recovery from SCI. It can down regulate the expression of Nogo-A and NgR protein.
2.Comparison of volatile constituents in two types of mugwort leaves (produced in Qichun and Nanyang) using the headspace GC-MS
Yuan ZHANG ; Liping KANG ; Zhongqiu TENG ; Zhilai ZHAN ; Tiegui NAN ; Aixiang ZHOU ; Lanping GUO
Journal of Acupuncture and Tuina Science 2016;14(3):164-169
Objective:To compare the volatile constituents in mugwort leaves produced in Qichun, Hubei Province and Nanyang, Henan Province.
Methods:The volatile constituents were extracted using headspace heating and analyzed using gas chromatography-mass spectrometry (GC-MS). Then a qualitative analysis was made according to the standard database provided by the National Institute of Standards and Technology (NIST) and the relative contents of each constituent were calculated using the peak area normalization method.
Results:A total of 59 compounds were identified from the mugwort leaves from Qichun and 51 compounds were identified from the mugwort leaves from Nanyang. These mainly include monoterpenoids, sesquiterpenoids, CxHyOz and other compounds involving the aldehyde, ketone, alkane and benzene. The mugwort leaves from Qichun and Nanyang share 32 common volatile constituents. The chromatographic peak area of identified compounds accounting for 96.38% of GC-MS total chromatographic peak areain Qichun mugwort leaves, versus 95.54% of that in Nanyang mugwort leaves.
Conclusion:The headspace heating extraction combined with GC-MS technology can evidently display similarities and differences of volatile constituents in mugwort leaves produced in different areas and thus provide scientific basis for the quality and screening of mugwort leaves.
3.Posterior fixation and fusion for treatment of Os odontoideum complicated by atlantoaxial dislocation.
Hui ZHANG ; Anmin JIN ; Li ZHANG ; Zhilai ZHOU ; Yang DUAN ; Shaoxiong MIN
Journal of Southern Medical University 2012;32(9):1358-1361
OBJECTIVETo summarize the techniques and evaluate the therapeutic effect of posterior fixation and fusion in the treatment of Os odontoideum complicated by atlantoaxial dislocation.
METHODSFrom March, 2007 to October, 2010, 10 patients with Os odontoideum (including 6 male and 4 female patients aged from 20 to 65 years, mean 39.8 years) were treated in our hospital. Before and after the operation, the patients underwent X ray, CT and MRI examinations to measure and evaluate the degree of dislocation and neural compression. After preoperative traction for 1-2 weeks, all the 10 patients showed deductible atlantoaxial dislocation. Through a posterior approach, Atlantoaxial pedicle screws fixation were performed in 9 cases, and C2/3 pedicle-Occiput screw fixation was performed in 1 case. All the patients wore cervical collars as external support for 3 months after the operation.
RESULTSThe mean operative time was 3 h in these patients with a mean intraoperative blood loss of 420 ml. The symptoms were relieved after the surgery in all the patients, who showed no neck pain or neurological defects. The patients were followed up for 6 to 52 months (mean 22 months), and bony fusion was observed in all the 10 cases within 6 to 8 months without such complications as internal fixation failure or redislocation of the atlas.
CONCLUSIONPatients with Os odontoideum complicated by atlantoaxial dislocation should undergo surgical stabilization to avoid severe neurological injury. Pedicle screw instrument in the atlas allows restoration of the spinal stability, short-segment fusion, and maximal preservation of the mobility of the neck.
Adult ; Aged ; Atlanto-Axial Joint ; injuries ; Bone Transplantation ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Joint Dislocations ; complications ; surgery ; Joint Loose Bodies ; complications ; surgery ; Male ; Middle Aged ; Odontoid Process ; pathology ; Spinal Fusion ; methods ; Treatment Outcome ; Young Adult
4.Rapid authentication of different herbal medicines by heating online extraction electrospray ionization mass spectrometry
Zidong QIU ; Chaofa WEI ; Xiang LI ; Changjiangsheng LAI ; Zhilai ZHAN ; Yan JIN ; Li ZHOU ; Qingxiu HAO ; Jian YANG ; Shuanglong WANG ; Liping KANG ; Luqi HUANG
Journal of Pharmaceutical Analysis 2023;13(3):296-304
The rapid and accurate authentication of traditional Chinese medicines(TCMs)has always been a key scientific and technical problem in the field of pharmaceutical analysis.Herein,a novel heating online extraction electrospray ionization mass spectrometry(H-oEESI-MS)was developed for the rapid and direct analysis of extremely complex substances without the requirement for any sample pretreatment or pre-separation steps.The overall molecular profile and fragment structure features of various herbal medicines could be completely captured within 10-15 s,with minimal sample(<0.5 mg)and solvent consumption(<20 μL for one sample).Furthermore,a rapid differentiation and authentication strategy for TCMs based on H-oEESI-MS was proposed,including metabolic profile characterization,characteristic marker screening and identification,and multivariate statistical analysis model validation.In an analysis of 52 batches of seven types of Aconitum medicinal materials,20 and 21 key compounds were screened out as the characteristic markers of raw and processed Aconitum herbal medicines,respectively,and the possible structures of all the characteristic markers were comprehensively identified based on Com-pound Discoverer databases.Finally,multivariate statistical analysis showed that all the different types of herbal medicines were well differentiated and identified(R2X>0.87,R2Y>0.91,and Q2>0.72),which further verified the feasibility and reliability of this comprehensive strategy for the rapid authentication of different TCMs based on H-oEESI-MS.In summary,this rapid authentication strategy realized the ultra-high-throughput,low-cost,and standardized detection of various complex TCMs for the first time,thereby demonstrating wide applicability and value for the development of quality standards for TCMs.
5.Herbal Textual Research on Olibanum in Famous Classical Formulas
Haiyan ZHOU ; Qingqing WANG ; Qi ZHANG ; Suping XIAO ; Meng CHEN ; Jianxin ZHOU ; Yeda ZHANG ; Danyang JIA ; Zhilai ZHAN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(4):55-66
By consulting the ancient and moderm literature, this paper makes a textual research on the name, origin, quality evaluation, harvesting and processing of Olibanum, so as to provide a basis for the development of the famous classical formulas containing this medicinal material. According to the herbal textual research, the results showed that Olibanum was first described as a medicinal material by the name of Xunluxiang in Mingyi Bielu(《名医别录》), until Ruxiang had been used as the correct name since Bencao Shiyi(《本草拾遗》) in Tang dynasty. The main origin was Boswellia carterii from Burseraceae family. The mainly producing areas in ancient description were ancient India and Arabia, while the modern producing areas are Somalia, Ethiopia and the southern Arabian Peninsula. The medicinal part of Olibanum in ancient and modern times is the resin exuded from the bark, which has been mainly harvested in spring and summer. It is concluded that the better Olibanum has light yellow, granular, translucent, no impurities such as sand and bark, sticky powder and aromatic smell. There were many processing methods in ancient times, including cleansing(water flying, removing impurities), grinding(wine grinding, rush grinding), frying(stir-frying, rush frying, wine frying), degreasing, vinegar processing, decoction. In modern times, the main processing methods are simplified to cleansing, stir-frying and vinegar processing. Nowadays, the commonly used specifications include raw, fried and vinegar-processed products. Among the three specifications, raw products is the Olibanum after cleansing, fried products is a kind of Olibanum processed by frying method, vinegar-processed products is the processed products of pure frankincense mixed with vinegar. Based on the research results, it is recommended to select the resin exuded from the bark of B. carterii for the famous classical formulas such as Juanbitang containing Olibanum, processing method should be carried out in accordance with the processing requirements of the formulas, otherwise used the raw products if the formulas without clear processing requirements.
6.Herbal Textual Research on Curculiginis Rhizoma in Famous Classical Formulas
Jinfang LUO ; Yaoyao HUANG ; Tao ZHOU ; Yang YU ; Guoqiong CAO ; Dan ZHAO ; Chenhong XIAO ; Zhilai ZHAN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(14):199-208
By consulting the ancient and modern literature, the name, origin, quality evaluation, harvesting and processing methods of Curculiginis Rhizoma(CR) were systematically combed and verified, in order to provide a basis for the development and utilization of famous classical formulas containing CR. The results of herbal textual research showed that the name Xianmao was first recorded in Leigong Paozhilun, the name of CR was used in all dynasties and this name came from its efficacy and morphological characteristics, the mainstream source for CR of the past dynasties was the rhizome of Curculigo orchioides or C. capitulata, since modern times, C. orchioides has been the main source of commodities. In ancient times, most of the places of origin of the description were the western regions and southwest China, while in modern times, Sichuan and Guizhou were regarded as genuine places. Since modern times, its quality has been summarized as the best with thick roots, firm texture and black-brown surface, the harvesting and processing methods recorded in the past dynasties are mainly sun drying after harvest in the second, eighth and ninth months of the lunar calendar, and most of them are harvested in autumn and winter in modern times. In ancient times, there were many processing methods of CR, mainly in processing with rice swill, while in modern times, stir-frying with wine was the main processing method. The nature, taste, meridian tropism, functions and indications of CR are basically consistent from ancient to modern times, the taboos for taking are to avoid iron, cow's milk, and beef. Although there are some differences in the understanding of the toxicity of CR in the past dynasties, most of the materia medica are clear that it has a certain toxicity. Based on the research conclusion, it is suggested that the rhizome of C. orchioides of Lycoris family should be used as its source in the famous classical formulas, and the corresponding processing method should be selected according to the processing requirements in the formulas, while the raw products is recommended to be selected as medicine if the processing requirement is not specified.
7.Herbal Textual Research on Eucommiae Cortex in Famous Classical Formulas
Qingqing WANG ; Haiyan ZHOU ; Xu ZHOU ; Qi ZHANG ; Danyang JIA ; Xingsong LU ; Suping XIAO ; Jianxin ZHOU ; Meng CHEN ; Zhilai ZHAN
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(19):88-96
In order to provide the basis for the development of famous classical formulas, the name, origin, quality evaluation, harvesting and processing of Eucommiae Cortex were systematically researched by consulting the ancient herbal and medical books, combining with the modern literature. According to the textual research, materia medica in the past dynasties used Eucommiae Cortex as the correct name. Combined with characteristics, origin and efficacy, Eucommiae Cortex in ancient times to the present is the dry bark of Eucommia ulmoides from family Eucommiaceae. The earliest producing areas of Eucommiae Cortex are Henan, Shanxi, Shaanxi and Sichuan. Since the Ming dynasty, the producing areas have expanded to most of the regions in the country, and Sichuan, Shaanxi, Chongqing, Guizhou and Hubei are regarded as the authentic producing areas. It has been concluded that the quality of Eucommiae Cortex is best if the bark has thick body, large block, scraped rough skin, multi silk section and dark purple internal surface. In ancient times, the processing methods of Eucommiae Cortex were mainly included removing rough bark and cutting for raw use, processing with auxiliary materials such as honey, ginger juice, salt water, wine, and so on. While in modern times, the processing methods have become increasingly simplified which are mainly cutting raw materials after cleansing and salt processing. It is need to excavate the connotation of different processed products and restore the traditional main processing methods through standards. Based on the requirement of Eucommiae Cortex in Sanbitang, it is suggested to use ginger-processed products according to the research results, which is used ginger juice as auxiliary material and processed with stir frying method according to the 2020 edition of Chinese Pharmacopoeia.
8.Herbal Textual Research on Bletillae Rhizoma in Famous Classical Formulas
Dan ZHAO ; Tao ZHOU ; Chaolei LUO ; Dewei GAN ; Lingling LIU ; Chuanzhi KANG ; Zhikun WU ; Xu LI ; Yan FU ; Guoqiong CAO ; Yongping ZHANG ; Chenghong XIAO ; Zhilai ZHAN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(4):77-88
In order to provide basic information for the utilization and development of famous classical formulas containing Bletillae Rhizoma, this article systematically analyzes the historical evolution of the name, origin, harvesting and processing of Bletillae Rhizoma by reviewing the ancient materia medica, prescription books, medical books and modern literature. The research results showed that Baiji(白及) was the main name, some scholars took Baiji(白芨) as its main name, and there were many other names such as Baiji(白给), Baigen(白根), Baiji(白苙). The mainstream source of Bletillae Rhizoma was the tubers of Bletilla striata, and drying, large, white, solid, root-free and skin removed completely were the good quality standards. With the promotion of wild to cultivated medicinal materials, there were certain differences between their traits, and the quality evaluation indexes should be adjusted accordingly. The origin of records in the past dynasties was widely distributed, with Guizhou and Sichuan having high production and good quality in modern times. The harvesting period is mostly in spring and autumn, and harvested in autumn was better. The processing and processing technology is relatively simple, and it was used fresh or powdered in past dynasties, while it is mainly sliced for raw use in modern times. Based on the results, it is suggested that the tubers of Bletilla striata of Orchidaceae should be used in the famous classical formulas, and it should be uniformly written as Baiji(白及). And if the original formula indicates the requirement of processing, it should be operated according to the requirement, if the requirement of processing is not indicated, it can be used in raw form as medicine.
9.Herbal Textual Research on Pruni Semen in Famous Classical Formulas
Baochang ZHOU ; Bin HE ; Ye LI ; Guoqing HAN ; Yaqiong BI ; Chunhong ZHANG ; Minhui LI ; Zhilai ZHAN
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(11):1-11
This article has systematically sorted out and verified the name, origin, producing area, quality evaluation, harvesting and processing of Pruni Semen by consulting ancient materia medica, medical books, prescription books and modern literature, in order to provide a basis for the development of famous classical formulas containing Pruni Semen. The results showed that Pruni Semen, as a medicinal material, has been widely used in medical literature of past dynasties since it was collected in Shennong Bencaojing, and also included under the names such as Yuhe, Yuzi and Yuli, and aliases such as Jueli, Queli and Chexiali. The primordial plants mentioned in the past dynasties involve about 12 species of Rosaceae, but with Prunus humilis, P. japonica and P. glandulosa as mainstream varieties used in the past dynasties, while the 2020 edition of Chinese Pharmacopoeia stipulates that the basal plants are P. humilis, P. japonica and P. pedunculata. Most of the ancient records for the origin of Pruni Semen are found everywhere in high mountains, valleys and hills, modern literature records that its origin varies according to its base, for example, P. humilis and P. japonica are mainly produced in Hebei, eastern Inner Mongolia, Liaoning, Shandong and other regions of China, and P. pedunculata is mainly produced in Inner Mongolia. Modern literature summarizes its quality as faint yellow, full and fulfilling, neat and not broken, and non-oiling, and the small Pruni Semen is better than the big Pruni Semen. The ancient processing methods of Pruni Semen mainly include blanching and peeling, blanching and peeling followed by frying, and blanching and peeling followed by pounding, with the common feature of blanching and peeling. The successive editions of Chinese Pharmacopoeia stipulate that it should be pounded when used. Based on the results of the herbal textual research and the writing time of Bianzhenglu, and combined with the market survey of Pruni Semen, it is suggested that P. humilis or P. japonica should be used as the origin of Pruni Semen in Sanpiantang, and it is harvested when the fruits are ripe, the kernels are collected by removing the stones, and processed by blanching, peeling and pounding consulting the decoction method in the current edition of Chinese Pharmacopoeia.
10.Comparison of Wild and Cultivated Paeoniae Radix Rubra Based on Traditional Quality Evaluation
Chunfang TIAN ; Qiannan HU ; Zhilai ZHAN ; Xiaoyan LAN ; Xiang LI ; Li ZHOU ; Tiegui NAN ; Zidong QIU ; Liping KANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(14):165-174
ObjectiveTo compare wild and cultivated Paeoniae Radix Rubra(PRR) in three aspects, including character, microscope, determination of primary and secondary metabolites. MethodSeventeen batches of wild and nine batches of cultivated PRR were collected,their character data were measured by vernier caliper and scales, and their paraffin sections were made by safranin-fixed green dyeing for the observation of microscopic features. The content of ethanol-soluble extracts and total tannin from wild and cultivated PRR was determined by the method of general principle 2201 and 2202 in the 2020 edition of Chinese Pharmacopoeia, the content of polysaccharides was determined by phenol-sulfuric acid method. Anthrone colorimetry was used to determine the content of starch, and Van Soest method of washing fiber was used to determine the content of fiber. The contents of fructose, glucose and sucrose in wild and cultivated PRR were determined by ultra-high performance liquid chromatography evaporative light scattering detection(UPLC-ELSD), and the secondary metabolites(gallic acid, methyl gallate, catechin, oxypaeoniflorin, albiflorin, paeoniflorin, ellagic acid, 1,3,4,6-tetragalloylglucose, galloylpaeoniflorin, 1,2,3,4,6-O-pentagalloylglucose, naringenin, benzoylpaeoniflorin and benzoylalbiflorin) were determined by UPLC. Principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were used to analyze the data of wild and cultivated PRR, the contribution of different factors to the difference was determined according to the variable importance in the projection(VIP) value>1 and P<0.05. ResultIn term of characters, wild PRR showed the traditional characteristic of Zaopi Fencha, its outer skin was loose and easy to fall off, its surface had longitudinal furrow and wrinkle, but the outer skin of cultivated PRR was not easy to fall off, and its surface was relatively smooth. The radial texture of xylem of wild PRR cross-section was more obvious, showing radial striations, vacuoles and more cracks, while the radial texture of xylem of cultivated PRR cross-section was not obvious, dense and some had cracks. Microscopically, the number of radial vessels arranged in the xylem of wild PRR was more than that of cultivated PRR, the number of calcium oxalate clusters in the phloem and xylem of wild PRR was more than that of cultivated PRR, while the number of starch grains was significantly higher in cultivated PRR. In terms of the content of primary chemical constituents, the contents of polysaccharides and starch of cultivated PRR were significantly higher than those of wild PRR(P<0.05), while the contents of cellulose, lignin, fructose and glucose of wild PRR were significantly higher than those of cultivated PRR(P<0.05). The results of determination of 13 secondary metabolites showed that the contents of paeoniflorin, methyl gallate, catechin and oxypaeoniflorin in wild PRR were significantly higher than those in cultivated PRR(P<0.05), while the contents of albiflorin, gallic acid, ellagic acid, naringenin, benzoylpaeoniflorin and benzoylalbiflorin were significantly lower than those of cultivated PRR(P<0.05). A total of 10 variables contributing to the differentiation between wild and cultivated PRR were screened, including albiflorin, cellulose, benzoylpaeoniflorin, oxypaeoniflorin, naringenin, ellagic acid, starch, lignin, paeoniflorin and total tannins. ConclusionThere are significant differences between wild and cultivated PRR in characters, microscopic characteristics, contents of primary and secondary metabolites. It is suggested that the content ratio of paeoniflorin and albiflorin, the contents of oxypaeoniflorin and cellulose can be used as indicators to characterize production methods of PRR so as to improve the quality standard of PRR. This study can provide reference for the improvement of quality standard of PRR and the guidance of high quality production of PRR.