[The rules of acupoint selection of acupuncture for dry eye].

The rules of acupoint selection of acupuncture for dry eye in recent 10 years were summarized to provide reference for clinical treatment. The clinical researches regarding acupuncture for dry eye published from May 5, 2007 to December 1, 2017 were searched in CNKI, WANFANG and VIP database. The acupoint and meridian used and application frequency were analyzed to summarize the rules of acupoint selection. Totally, 52 related clinical papers were collected, involving 60 acupoints, 14 meridians and 5 extra point. The total frequency of acupoints was 541, which were mainly related with the bladder meridian, stomach meridian and gallbladder meridian. In conclusion, the rules of acupoint selection of acupuncture for dry eye highlighted meridians circulation, acupoint combination, and syndrome differentiation.

[Effect of Warm Acupuncture on the Levels of Serum Immunoglobulin E, Interleukin-1 ß and Tumor Necrosis Factor-α in Rats with Allergic Rhinitis].

OBJECTIVE: To observe the effect of warm acupuncture on behavior and contents of serum immunoglobulin E(IgE), interleukin-1 ß(IL-1 ß) and tumor necrosis factor-α(TNF-α) in allergic rhinitis(AR) rats, so as to explore its mechanism underlying improving AR. METHODS: Forty Wistar rats were randomly divided into four groups: control group, model group, medication group and warm acupuncture group(10 rats/group). The AR model was established by intraperitoneal injection of sensitization and nasal drip. The rats in the medication group were given fluticasone propionate nasal spray, daily for 10 days. Warm acupuncture was applied to "Fengchi"(GB 20), "Yintang"(GV 29), "Yingxiang"(LI 20) for 60 seconds, once daily for 10 days. Behavioral scores were used to evaluate behavioral changes in rats. Enzyme linked immunosorbent assay (ELISA) was used to detect the expression levels of serum IgE, IL-1 ß and TNF-α. RESULTS: Behavioral scores of the model group were significantly higher than those of the control group 0, 3, 7 and 10 days after modeling (P<0.05). After treatment, the behavioral scores of medication group and the warm acupuncture group were lower than those of the model group (P<0.05), and the score was more lower in the warm acupuncture group than in the medication group (P<0.05). Compared with the control group, the levels of serum IgE, IL-1 ß and TNF-α in the model group were all increased (P<0.01), while the levels of serum IgE, IL-1 ß and TNF-α were decreased in the medication and warm acupuncture groups after treatment in comparison with the model group (P<0.05, P<0.01). Compared with the medication group, the levels of serum IgE, IL-1 ß and TNF-α were significantly lower in the warm acupuncture group (P<0.05, P<0.01). CONCLUSION: The expression levels of IgE, IL-1 ß and TNF-α were elevated in serum after AR attack. Warm acupuncture can improve the symptoms of AR rats, which may be associated to its effect in inhibiting the expression of serum IgE, IL-1 ß and TNF-α.

Research on human glioma stem cells in China.

Research on human glioma stem cells began early in the 21st century and since then has become a rapidly growing research field with the number of publications increasing year by year. The research conducted by our diverse group of investigators focused primarily on cell culture techniques, molecular regulation, signaling pathways, cancer treatment, the stem cell microenvironment and the cellular origin and function of glioma stem cells. In particular, we put forward our view that there are inverse or forward transformations among neural stem cells, glial cells and glioma stem cells in glioma tissues under certain conditions. Based on the background of the progress of international research on human glioma stem cells, we aim to share our progress and current findings of human glioma stem cell research in China with colleagues around the world.

The establish of the HPLC method to examine the plasma concentration of lamotrigine and oxcarbazepine.

To establish the HPLC method to examine plasma concentration of lamotrigine and oxcarbazepine. This study set chlorzoxazone as the internal standard, chromatographic column was Column C18 (200×4.6mm, 5um) of DIKMA company, the mobile phase was methanol, water and trifluoroacetic acid, with rate of 40: 60: 0.0005, at a flow rate of 1 mllmin(-1), the detected wavelength was 240 nm. The plasma concentrations of lamotrigine was 0.5-50ug•mL(-1), the standard curve was excellent for Y=0.5511C-0.5669, r=0.9940, average recovery was 91.40%; The plasma concentrations of oxcarbazepine was 0.5-50ugmL-1, the standard curve was good for Y=0.4026C-0.5895, r=0.9925, and the average recovery was 89.59%; The three plasma concentrations of lamotrigine were respectively 25µg•mL(-1), 10 µg•mL(-1) and 2µg•mL(-1) and its five parallel sample for injection RSD were respectively 4.01%, 6.15% and 4.64%; The three plasma concentration of oxcarbazepine were 25µg•mL(-1)-1(-1), 10µg•mL(-1)-1(-1) and 2µg•mL(-1)-1(-1), and its five parallel sample for injection RSD were respectively 3.05%, 4.27% and 9.01%. This method was easy to operate, high recovery and high precision, and was applicable to the clinical detection for plasma concentration of lamotrigine and oxcarbazepine.

[Warming-promotion acupuncture for post-earthquake depression: a randomized controlled study].

OBJECTIVE: To observe the clinical efficacy differences among warming-promotion acupuncture, regular acupuncture and psychological care for post-earthquake depression. METHODS: Ninety cases of post-earthquake depression in Gansu province were randomly divided into a warming-promotion acupuncture group, a regular acupuncture group and a care group, 30 cases in each one. The psychological treatment was applied in the care group; the regular acupuncture at Baihui (GV 20), Sishencong (EX-HN 1)), Fengchi (GB 20), Shenmen (HT 7), Yang-lingquan (GB 34), Taichong (LR 3) and Sanyinjiao (SP 6) was used in the regular acupuncture group; based on the treatment of regular acupuncture group, the warming-promotion acupuncture was applied in the warming-promotion acupuncture group, once a day. Ten days were taken as one course with an interval of two days between two courses. Totally 3 treatment courses were completed. The Hamilton depression (HAMD) scale before and after treatment in each group was observed while the efficacy was analyzed. RESULTS: All three treatments were effective for depression. The total cured rate was 86.67% (26/30) in the warming-promotion acupuncture group, which was obviously superior to 50.00% (15/30) in the regular acupuncture group (P < 0.01) and 33.33% (10/30) in the care group (P < 0.001). After the treatment, the score of HAMD was all reduced in three groups, in which the effect in warming-promotion acupuncture group was superior to that in the regular acupuncture group and care group (9.02 +/- 2.37 vs 10.73 +/- 2.28 and 11.05 +/- 2.01, P < 0.05, P < 0.01). CONCLUSION: The warming-promotion acupuncture has positive effect on post-earthquake depression, which has special effect and is worthy of clinical generalization and application.

[Clinical observation on two-way quintuple puncture in the treatment of ganglion].

OBJECTIVE: To explore the better therapy in the treatment of ganglion. METHODS: Ninety cases of ganglion were randomized into a two-way quintuple puncture group, a common quintuple puncture group and a fire needling group, 30 cases in each one. In the two-way quintuple puncture group, the "9-in-1" multiple penetrating needling technique was used. In the common quintuple puncture group, the traditional "5-in-1" multiple penetrating needling technique was applied. In the fire needling group, the traditional multiple fire needling technique was adopted. The treatment was given once a day, 3 treatments made one session and the efficacy was analyzed statistically after 1 session treatment in the three groups. RESULTS: All of the three therapeutic methods achieved the efficacy on ganglion. The curative rate was 96. 7% (29/30) in the two-way quintuple puncture group, which was better obviously than 66.7% (20/30) in the common quintuple puncture group and 60. 0% (18/30) in the fire needling group (both P<0. 01). CONCLUSION: The two-way quintuple puncture technique achieves the remarkably superior efficacy on ganglion as compared with the common quintuple puncture technique and fire needling technique.

[Host glial cell canceration induced by glioma stem cells in GFP/RFP dual fluorescence orthotopic glioma models in nude mice].

OBJECTIVE: During the process of tissue remodeling in human tumor transplantation models, the roles of the inoculated tumor cells and host tissue in tumor progression is still largely unknown. The aim of this study was to investigate the relationships and interactions between these two sides using GFP-RFP double fluorescence tracing technique. METHODS: Red fluorescence protein (RFP) gene was stably transfected into glioma stem cell line SU3, then SU3-RFP cells were transplanted into the brain of athymic nude mice with green fluorescence protein (GFP) expression. After the intracerebral tumors were formed, the relationship and interaction between GFP cells and RFP cells were analyzed. Highly proliferative GFP cells were screened out, and monocloned with micro-pipetting. DNA content assay, chromosome banding and carcinogenicity test of the GFP cells were performed to observe the GFP cells' cancerous phenotype in nude mice. RESULTS: In the transplantable tumor tissue, besides a great quantity of RFP cells, there were still a proportion of GFP cells and GFP/RFP fusion cells. The proportion of RFP cells, GFP cells and GFP/RFP cells were (88.99 ± 1.46)%, (5.59 ± 1.00)%, and (4.11 ± 1.020)%, respectively. Two monoclonal host GFP cells (H1 and H9) were cloned, which demonstrated the properties of immortality, loss of contact inhibition, and ultra-tetraploid when cultured in vitro. Both H1 and H9 cells expressed CNP, a specific marker of oligodendrocytes. The GFP cells also demonstrated 100% tumorigenic rate and high invasive properties in vivo. CONCLUSIONS: In this glioma transplantation model, the transplanted tumor tissues contained not only transplanted glioma stem cells but also cancerous host GFP cells. Our findings offer important clues to further research on the relationships among different members in the tumor microenvironment.

[Clinical observation on warming-promoting needling and target penetrating needling method for chronic prostatitis].

OBJECTIVE: To explore a more optimal therapy for chronic prostatitis. METHODS: Ninty cases of chronic prostitis were randomized into a warming-promoting needling and target penetrating needling method group (WT group), a conventional acupuncture group (CA group) and a medication group (M group), 30 cases in each one. The warming-promoting needling method was applied at Zhongji (CV 3), Guanyuan (CV 4), Qihai (CV 6), Daheng (SP 15), Qichong (ST 30) in the WT group, the needling sensation was targeted towards the prostate, once daily; in CA group, conventional acupuncture was applied at the same acupoints of WT group; Qianliekang tablets was prescribed to the M group. For the 3 groups, 10 treatment made one session and 3 days as an interval, the efficacy was evaluated after 3 sessions. RESULTS: All of the 3 methods were effective for chronic prostitis, the total effective rate in the WT group was 93.3% (28/30), which was superior to 73.3% (22/30, P < 0.05) in the CA group and 63.3% (19/30, P < 0.01) in the M group. The curative rate in the WT group was 80.0% (24/30), which was superior to 46.6% (14/30) in the CA group and 30.0% (9/30) in the M group (both P < 0.001). CONCLUSION: The warming-promoting needling and target penetrating needling method achieve superior efficacy on chronic prostatitis as compared with conventional acupuncture and Qianliekang tablets.

Incubation and application of transgenic green fluorescent nude mice in visualization studies on glioma tissue remodeling.

BACKGROUND: The primary reasons for local recurrence and therapeutic failure in the treatment of malignant gliomas are the invasion and interactions of tumor cells with surrounding normal brain cells. However, these tumor cells are hard to be visualized directly in histopathological preparations, or in experimental glioma models. Therefore, we developed an experimental human dual-color in vivo glioma model, which made tracking solitary invasive glioma cells possible, for the purpose of visualizing the interactions between red fluorescence labeled human glioma cells and host brain cells. This may offer references for further studying the roles of tumor microenvironment during glioma tissue remodeling. METHODS: Transgenic female C57BL/6 mice expressing enhanced green fluorescent protein (EGFP) were crossed with male Balb/c nude mice. Then sib mating was allowed to occur continuously in order to establish an inbred nude mice strain with 50% of their offspring that are EGFP positive. Human glioma cell lines U87-MG and SU3 were transfected with red fluorescent protein (RFP) gene, and a rat C6 glioma cell line was stained directly with CM-DiI, to establish three glioma cell lines emitting red fluorescence (SU3-RFP, U87-RFP, and C6-CM-DiI). Red fluorescence tumor cells were inoculated via intra-cerebral injection into caudate nucleus of the EGFP nude mice. Tumor-bearing mice were sacrificed when their clinical symptoms appeared, and the whole brain was harvested and snap frozen for further analysis. Confocal laser scanning microscopy was performed to monitor the mutual interactions between tumor cells and host brain cells. RESULTS: Almost all the essential tissues of the established EGFP athymic Balb/c nude mice, except hair and erythrocytes, fluoresced green under excitation using a blue light-emitting flashlight with a central peak of 470 nm, approximately 50% of the offsprings were nu/nu EGFP+. SU3-RFP, U87-RFP, and C6-CM-DiI almost 100% expressed red fluorescence under the fluorescence microscope. Under fluorescence microscopic view, RFP+ cells were observed growing wherever they arrived at, locating in the brain parenchyma, ventricles, and para-vascular region. The interactions between the transplanted tumor cells and host adjacent cells could be classified into three types: (1) interweaving; (2) mergence; and (3) fusion. Interweaving was observed in the early stage of tumor remodeling, in which both transplantable tumor cells and host cells were observed scattered in the tumor invading and spreading area without organic connections. Mergence was defined as mutual interactions between tumor cells and host stroma during tumorigenesis. Direct cell fusion between transplantable tumor cells and host cells could be observed occasionally. CONCLUSIONS: This study showed that self-established EGFP athymic nude mice offered the possibility of visualizing tumorigenesis of human xenograft tumor, and the dual-color xenograft glioma model was of considerable utility in studying the process of tumor remodeling. Based on this platform, mutual interactions between glioma cells and host tissues could be observed directly to further elucidate the development of tumor microenvironment.

Glioma stem cells are more aggressive in recurrent tumors with malignant progression than in the primary tumor, and both can be maintained long-term in vitro.

BACKGROUND: Despite the advances made during decades of research, the mechanisms by which glioma is initiated and established remain elusive. The discovery of glioma stem cells (GSCs) may help to elucidate the processes of gliomagenesis with respect to their phenotype, differentiation and tumorigenic capacity during initiation and progression. Research on GSCs is still in its infancy, so no definitive conclusions about their role can yet be drawn. To understand the biology of GSCs fully, it is highly desirable to establish permanent and biologically stable GSC lines. METHODS: In the current study, GSCs were isolated from surgical specimens of primary and recurrent glioma in a patient whose malignancy had progressed during the previous six months. The GSCs were cryopreserved and resuscitated periodically during long-term maintenance to establish glioma stem/progenitor cell (GSPC) lines, which were characterized by immunofluorescence, flow cytometry and transmission electronic microscopy. The primary and recurrent GSPC lines were also compared in terms of in vivo tumorigenicity and invasiveness. Molecular genetic differences between the two lines were identified by array-based comparative genomic hybridization and further validated by real-time PCR. RESULTS: Two GSPC lines, SU-1 (primary) and SU-2 (recurrent), were maintained in vitro for more than 44 months and 38 months respectively. Generally, the potentials for proliferation, self-renewal and multi-differentiation remained relatively stable even after a prolonged series of alternating episodes of cryopreservation and resuscitation. Intracranial transplantation of SU-1 cells produced relatively less invasive tumor mass in athymic nude mice, while SU-2 cells led to much more diffuse and aggressive lesions strikingly recapitulated their original tumors. Neither SU-1 nor SU-2 cells reached the terminal differentiation stage under conditions that would induce terminal differentiation in neural stem cells. The differentiation of most of the tumor cells seemed to be blocked at the progenitor cell phase: most of them expressed nestin but only a few co-expressed differentiation markers. Transmission electron microscopy showed that GSCs were at a primitive stage of differentiation with low autophagic activity. Array-based comparative genomic hybridization revealed genetic alterations common to both SU-1 and SU-2, including amplification of the oncogene EGFR and deletion of the tumor suppressor PTEN, while some genetic alterations such as amplification of MTA1 (metastasis associated gene 1) only occurred in SU-2. CONCLUSION: The GSPC lines SU-1 and SU-2 faithfully retained the characteristics of their original tumors and provide a reliable resource for investigating the mechanisms of formation and recurrence of human gliomas with progressive malignancy. Such investigations may eventually have major impacts on the understanding and treatment of gliomas.

[Ultrastructural analysis of glioma stem cells-progenitors].

OBJECTIVE: It is well known that glioma stem cells-progenitors (GSCP) proliferate indefinitely and hardly differentiate in vitro, however, the reasons remain unknown. The aim of this study was to explore the ultrastructural basis of GSCP. METHODS: GSCP, kept by our laboratory, were collected, embedded, and cut into ultrathin sections and observed under the transmission electron microscope. RESULTS: A single GSCP usually had relatively well developed mitochondria, Golgi apparatuses, ribosomes, and undeveloped rough endoplasmic reticulum, but seldom lysosomes and no typical autophagosomes were found, and the nuclear-cytoplasmic ratio was high. The nuclei frequently contained huge amounts of euchromatin and a small quantity of heterochromatin, and in most nuclei there were only one nucleolus, however, two or more nucleoli were also common. Typical apoptotic cells could hardly be found in tumor-spheres, and between neighboring cells in tumor-spheres there were incompletely developed desmosomes or intermediate junction. CONCLUSION: The ultrastructural features of glioma stem cells-progenitors showed that BTSCP were very primitive and the lack of autophagy and the underdevelopment of some other cellular organelles are probably the reasons for the differential inhibition of GSCPs.

[Treatment of 60 cases of allergic rhinitis mainly with point-through-point method].

OBJECTIVE: To search for an effective therapy for allergic rhinitis. METHODS: One hundred and twenty cases of allergic rhinitis were randomly divided into a treatment group (n=60) treated mainly with penetration needling, Yintang (EX-HN 3)-through-Bigen (root of nose), Sibai (ST 2)-through-Bigen, Yingxiang (LI 20)-through-Bigen, and a control group (n=60) treated with oral Biyankang, 4 tablets each time, 3 times daily. Their therapeutic effects were compared. RESULTS: The total effective rate was 85.0% in the treatment group and 60.0% in the control group, with a very significant difference between the two groups (P<0.01). CONCLUSION: The penetration needling as main therapy has a definite therapeutic effect on allergic rhinitis, which is significantly better than that of oral Biyankang.