[Exploration of the construction of semantic framework of meridians and acupoints based on top-level ontology].

Based on the top-level ontology and the existing ontology methodology, the related concepts of meridians and acupoints were discriminated, defined and classified; the relationship of core concepts were established, e.g. meridians, acupoints and zangfu. It was attempted to build an ontological semantic framework of meridians and acupoints. Through the investigation on the classification mode of the top-level ontology, it is proposed that the meridians and acupoints, as the unique concepts of traditional Chinese medicine, exist in the form of "emptiness" and belong to "immaterial entity". Meridians refer to the three-dimensional channels in the human body, and acupoints are divided into ontological acupoints and body surface ones. Ontological acupoints are regarded as a three-dimensional structure within the human body, whereas, body surface ones are the optimal sites for acupuncture needle insertion on the body surface, meaning, the zero-dimensional point on the body surface. The main relationships between meridians and acupoints include is-a, exterior-interior, located-in, correspondent-to, mapping, etc. The exploration of the semantic framework of meridians and acupoints is conductive to understanding the connotation of meridians, acupoints and their relationship.

The optimum parameters and neuroimaging mechanism of repetitive transcranial magnetic stimulation to post-stroke cognitive impairment, a protocol of an orthogonally-designed randomized controlled trial.

OBJECTIVE: Repetitive Transcranial Magnetic Stimulation (rTMS) has been used in cognition impairment due to various neuropsychiatric disorders. However, its optimum parameters and the neuroimaging mechanism are still of uncertainty. In order to simulate a study setting as close to real world as possible, the present study introduces a new orthogonally-designed protocol, consisting of the rTMS intervention with four key parameters (stimulating site, frequency, intensity and pulse number) and three different levels in each one, and aims to investigate the optimum parameters and the brain activity and connectivity in default mode network (DMN), dorsal attention network (DAN), central executive network (CEN) following rTMS intervention to post-stroke cognition impairment (PSCI). METHODS: A single-center, orthogonally-designed, triple-blind randomized controlled trial will be conducted and forty-five PSCI patients will be recruited and randomly assigned to one of nine active rTMS groups based on four rTMS paraments: stimulating site, frequency, intensity and pulse number. Neuropsychological, activities of daily living, quality of life and functional magnetic resonance imaging (fMRI) evaluations were be performed pre-, post- and 3 months after rTMS. DISCUSSION: This study evaluates the optimum parameters of rTMS for patients with post-stroke cognition impairment and explores the alteration of neural function in DMN, DAN, CEN brain network. These results would facilitate the standardized application of rTMS in cognition impairment rehabilitation.

Miniature Fourier transform spectrometer based on a fiber-tip interferometer.

The miniaturization of spectrometers have attracted much attention owning to the demand for portable or in situ spectral analysis in a wide variety of fields, but it is a great challenge to push them into practical applications due to high cost, complicated configuration, and sensitivity to external disturbance. We report on a miniature Fourier transform (FT) spectrometer based on fiber-tip Fizeau interferometer. Hand pulling or any other types of force can be used to drive optical path difference (OPD) scan. Interferences are monitored as a function of time by two photodetectors, one is used to detect the whole interferogram while the other to measure single-wavelength interferogram. In this design, the instantaneous interference intensity as well as OPD can be obtained in an accurate way so that the exact spatial interferogram of the incident spectrum can be worked out. Consequently, the incident spectrum can be retrieved by FT method. A resolution of 7.69 cm-1 in the wavelength range of 1400 nm ∼ 1700 nm is achieved. Experimental results show that the performance of our device is comparable to the commercial benchtop spectrometer. Our device is independent of the complicated fabrication procedures, easy of usage, and cost effective. We envision that the proposed design will inspire a new concept for constructing simple and cheap spectrometers that is well suited for practical applications.

A miniature fiber-optic microphone based on plano-concave micro-interferometer.

The sensitive detection of sound waves is essential for a variety of applications. In this work, we propose a miniature diaphragm-free fiber-optic microphone based on a plano-concave optical micro-interferometer. A solid plano-concave micro-interferometer is formed at the end of a cleaved fiber by depositing a tiny volume of liquefied glass. Sound wave induced periodic variation of pressure can significantly modify the refractive index of the plano-concave glass due to the elasto-optic effect, and then, the phase difference between two interferometric beams will be remarkably changed accordingly. The interferometer finally converts the fluctuation of the phase difference into the change in the output optical power. Consequently, the sound wave can be demodulated by detecting the output power of the microphone. The experimental results show that the proposed microphone has the ability to detect sound waves in the whole audible range and almost omnidirectional. The noise-limited minimum detectable sound pressure is around 12 µPa/Hz. In addition, the human voice detection test shows that the performance of our microphone is competitive with the most advanced commercial device. The structure is stable without any movable mechanical parts, and the size is as small as 0.25 mm, which makes the proposed microphone an attractive alternative to the conventional one for sound wave detection.

[The ancient literature source and flow of the point to point penetration acupuncture method].

It has been the consensus of the academic community that the doctor DOU Han-qing in Jin-Yuan Dynasty was the pioneer of point to point penetration acupuncture method.The contents of several texts in Yuan-Ming-Qing Dynasties were combed on the basis of Dou Taishi Zhenjing(《》).The paradigm,the acupoints,basic elements and the characteristics of the main treatment of diseases were discussed,and the relationship between the point to point penetration acupuncture method with the penetration acupuncture method has been tried to distinguish from the perspective of technical philosophy. It was found that the specifications and quantity of the 28 "penetration and to" cases in Dou Taishi Zhenjing(《》)were sufficiently exemplary for future generations, and were the source documents of Yuan-Ming-Qing's texts,which basically followed its specifications and content and developed.For example,the Yulong Poetries(《》)in the books of Bianque Shenying Zhenjiu Yulong Jing(《》),Zhenjiu Dacheng(《》),Zhenfang Liuji(《》).The 37 cases in Zhenfang Liuji(《》) were the most in several texts.The 31 cases in Xunjing Kaoxue Bian(《》)had the characteristics of DOU Han-qing, YANG Ji-zhou and LING Yun, who developed the opposite direction penetration method with two or more needles on DOU Han-qing's one needle.There were five basic elements in the point to point penetration acupuncture method:the needle pri-cking inch(depth), the needle pricking angle, the needle pricking direction, the starting acupoint, the end acupoint.This method in Dou Taishi Zhenjing(《》)mostly treated local diseases and symptoms,some acupoints below the elbows and knees also treated far-end diseases.The point to point penetration acupuncture method has a specific intention structure, which cannot be confused with the penetration acupuncture method.

Cerebellar Theta Burst Stimulation on Walking Function in Stroke Patients: A Randomized Clinical Trial.

Objectives: The objective of this study was to explore the efficacy of cerebellar intermittent theta burst stimulation (iTBS) on the walking function of stroke patients. Methods: Stroke patients with walking dysfunction aged 25-80 years who had suffered their first unilateral stroke were included. A total of 36 patients [mean (SD) age, 53 (7.93) years; 10 women (28%)] were enrolled in the study. All participants received the same conventional physical therapy, including transfer, balance, and ambulation training, during admission for 50 min per day during 2 weeks (10 sessions). Every session was preceded by 3 min procedure of cerebellar iTBS applyed over the contralesional cerebellum in the intervention group or by a similar sham iTBS in control group. The groups were formed randomly and the baseline characteristics showed no significant difference. The primary outcome measure was Fugl-Meyer Assessment-Lower Extremity scores. Secondary outcomes included walking performance and corticospinal excitability. Measures were performed before the intervention beginning (T0), after the first (T1) and the second (T2) weeks. Results: The Fugl-Meyer Assessment for lower extremity scores slightly improved with time in both groups with no significant difference between the groups and over the time. The walking performance significantly improved with time and between group. Two-way mixed measures ANOVA showed that there was significant interaction between time and group in comfortable walking time (F 2,68 = 6.5242, P = 0.0080, η2 partial = 0.276, ε = 0.641), between-group comparisons revealed significant differences at T1 (P = 0.0072) and T2 (P = 0.0133). The statistical analysis of maximum walking time showed that there was significant interaction between time and groups (F 2,68 = 5.4354, P = 0.0115, η2 partial = 0.198, ε = 0.734). Compared with T0, the differences of maximum walking time between the two groups at T1 (P = 0.0227) and T2 (P = 0.0127) were statistically significant. However, both the Timed up and go test and functional ambulation category scale did not yield significant differences between groups (P > 0.05). Conclusion: Our results revealed that applying iTBS over the contralesional cerebellum paired with physical therapy could improve walking performance in patients after stroke, implying that cerebellar iTBS intervention may be a noninvasive strategy to promote walking function in these patients. This study was registered at ChiCTR, number ChiCTR1900026450.

Cerebral Small Vessel Disease Load Predicts Functional Outcome and Stroke Recurrence After Intracerebral Hemorrhage: A Median Follow-Up of 5 Years.

Background: Uncertainty exists over the long-term prognostic significance of cerebral small vessel disease (CSVD) in primary intracerebral hemorrhage (ICH). Methods: We performed a longitudinal analysis of CSVD and clinical outcomes in consecutive patients with primary ICH who had MRI. Baseline CSVD load (including white matter hyperintensities [WMH], cerebral microbleeds [CMBs], lacunes, and enlarged perivascular spaces [EPVS]) was evaluated. The cumulative CSVD score was calculated by combining the presence of each CSVD marker (range 0-4). We followed participants for poor functional outcome [modified Rankin scale [mRS] ≥ 4], stroke recurrence, and time-varying survival during a median follow-up of 4.9 [interquartile range [IQR] 3.1-6.0] years. Parsimonious and fuller multivariable logistic regression analysis and Cox-regression analysis were performed to estimate the association of CSVD markers, individually and collectively, with each outcome. Results: A total of 153 patients were included in the analyses. CMBs ≥ 10 [adjusted OR [adOR] 3.252, 95% CI 1.181-8.956, p = 0.023] and periventricular WMH (PWMH) (adOR 2.053, 95% CI 1.220-3.456, p = 0.007) were significantly associated with poor functional outcome. PWMH (adOR 2.908, 95% CI 1.230-6.878, p = 0.015) and lobar CMB severity (adOR 1.811, 95% CI 1.039-3.157, p = 0.036) were associated with stroke recurrence. The cumulative CSVD score was associated with poor functional outcome (adOR 1.460, 95% CI 1.017-2.096) and stroke recurrence (adOR 2.258, 95% CI 1.080-4.723). Death occurred in 36.1% (13/36) of patients with CMBs ≥ 10 compared with 18.8% (22/117) in those with CMB < 10 (adjusted HR 2.669, 95% CI 1.248-5.707, p = 0.011). In addition, the cumulative CSVD score ≥ 2 was associated with a decreased survival rate (adjusted HR 3.140, 95% CI 1.066-9.250, p = 0.038). Conclusions: Severe PWMH, CMB, or cumulative CSVD burden exert important influences on the long-term outcome of ICH.

[Effects of Trunk Control Training on Dynamic Sitting Balance and Trunk Function in Hemiplegia Patients after Acute Stroke].

OBJECTIVE: To investigate the effect of trunk control training on trunk function, balance, mobility and functional independence in hemiplegia patients after acute stroke, and to compare the therapeutic effects with conventional stroke rehabilitation. METHODS: The study was designed as a randomized controlled trial. The subjects and evaluators were blinded and the therapist was unblinded. Subjects were randomly assigned to either the experimental group or the control group using a computer-generated random number table, the experimental group ( n=15) received30 min of high-intensity trunk control training plus 15 min of low-intensity conventional stroke rehabilitation (45 min, once1 d for 5 d). The control group ( n=15) received only 45 min low-intensity conventional stroke rehabilitation, once 1 d for 5 d. Trunk function was the primary outcome, which was evaluated by the Trunk Impairment Scale (TIS). The secondary outcome included balance, mobility and functional independence, which were assessed by the Brunel Balance Assessment (BBA), the Modified Rivermead Mobility Index (MRMI) and the modified Barthel Index (MBI). Subjects were assessed before and after the intervention. RESULTS: After 5 d of treatment, the scores of TIS general table, TIS static sitting balance sub table, TIS coordination sub table, BBA, MRMI and MBI were increased, but there was no statistically significant difference between the two groups (all P>0.05). The trunk control training group improved dynamic sitting balance more significantly, the score of TIS dynamic sitting balance sub table increased more than that in the control group, with a statistically significant difference ( P<0.05). CONCLUSION: The results of this study indicate that trunk control training has improved trunk function, balance, mobility, and activities of daily living in hemiplegia patients after acute stroke, and trunk control training improves dynamic sitting balance more effectively than conventional stroke rehabilitation.

[Collection methods, stylistic layout and value of extraordinary points in Doutaishi Zhenjing (Master Dou's Canon of Acupuncture)].

The literature source, collection methods and stylistic layout, value and influence of 32 extraordinary points in Doutaishi Zhenjing (Master Dou's Canon of Acupuncture) were collected. The sources of the literature are extensive. The collection methods include 5 aspects, named inheritance, integration and repair; name rectification, identification and differentiation; new point explication on the base of extraordinary points; well arrangement of points and mutual use of three methods for stylistic layout; as well as integrative illustration with text and picture, and determined location. In reference to the holistic layout of name, location, techniques of acupuncture and moxibustion and indications of extraordinary points in Doutaishi Zhenjing (Master Dou's Canon of Acupuncture), it is the first time to make a standardized and unified collection of extraordinary points, which is valuable in the historical literature, the theoretic research and clinical practice of extraordinary points. It brings a great impact on the special articles or books after the Yuan dynasty and even the establishment of national standards at present.

[Evolution of the concept of extra ordinary point and standardization of related terms].

Through an analysis of the historical development of the concept of extra ordinary point, it is found that the terms of "Qi point" "extra ordinary point" "Loujing point" "extra meridian point" and "Bie point" used in the ancient medical books in the Yuan, Ming, and Qing Dynasties actually expressed two different concepts of "extra ordinary points with the magical effects" and "extra points other than the meridian points". The standardized term for the former concept should be "extra ordinary point" (Qi point for short), and the standardized term for the latter concept should be "extra meridian point", which is also called "Loujing point" and "Bie point". Since these two different concepts are not clarified in modern acupuncture textbooks, especially the misreading of "Jing Wai", no consensus has been reached on the definition of "extra ordinary point" and "extra meridian point", and the standardized terms have not been decided, which changed from "extra ordinary point" to "Qi point" and "extra point" and recently returned to "extra ordinary point". These confusions in textbooks have a direct impact on the determination of the related acupuncture terms used in national and international standards.

[Relationship between DOU Han-qing of the Jin and Yuan Dynasty and Taoist].

In order to explore the culture factors of the construction of acupuncture theory of DOU Han-qing, the medical master of the Jin and Yuan Dynasty, the authors studied the relationship between DOU Han-qing and Taoist by collecting DOU 's life material. With manual retrieval, some ancient historical literature was obtained. Additionally, Yijing Huiyuan: Zhenjiu Yuanshu: Dou Taishi Michuan Mihua Zhenjing Qiongyao Baojian, written by WU Jia-yan in the Ming Dynasty, collected in Japan, was analyzed. It was found that DOU Han-qing's acupuncture and moxibustion was influenced by Suwen Wangpian: Cifalun, the Taoism medicine of the early stage of the Song Dynasty, as well as by Jiaojing Baxue, passed on by SONG Zi-hua, the hermit. DOU Han-qing had been in contact with FAN Zhi-ying of the Taoist, but there was no clear record relevant with medicine. Dou Taishi Michuan Mihua Zhenjing Qiongyao Baojian may be the compilation by Taoism medical master, on the base DOU 's acupuncture and moxibustion. There is a kind of mutual influence and mutual promotion relationship between DOU Han-qing and Taoist.

[Objective, approach and symbolic achievement of the standardization on extraordinary points].

Through sorting out the development of meridian points and the standardization of extraordinary points in dynasties, two objectives are discovered in the standardization of extraordinary points starting from the Tang Dynasty. The points with extraordinary effect are selected points and transferred to meridian points and the commonly-used key points are selected and developed to be standardized acupoints. Hence, 3 approaches are summarized. Approach No.1: a large number of "meridian-outside" points that are the potential supplementation but not included yet in meridian points are supplemented to meridian points. Approach No.2: based on the specified requirement, the key points are selected from the acupoints with determined locations and names, which is developed to be the standard of acupoints. Approach No.3: the acupoints with extraordinary effect are selected from the "meridian-outside" points and these points are collected for the formulation of the standard of extraordinary points. The latter two approaches provide for the development direction of teaching and standardization in the future.

X4-Tropic Latent HIV-1 Is Enriched in Peripheral Follicular Helper T Cells and Is Correlated with Disease Progression.

Follicular helper T (TFH) cells have been shown to support productive human immunodeficiency virus type 1 (HIV-1) replication and to serve as a key component of the latent viral reservoir. However, the viral characteristics of this latent reservoir and the clinical relevance of this reservoir remain unclear. In this study, we assessed the tropic composition of latent viruses from peripheral TFH (pTFH), non-TFH memory, and naive CD4+ T cells from individuals with HIV-1 infections on suppressive combined antiretroviral therapy (cART). X4-tropic latent HIV-1 was preferentially enriched in pTFH cells compared to levels in the other two subsets. Interestingly, the ratio of X4-tropic latent HIV-1 in pTFH cells not only was robustly and inversely correlated with blood CD4+ T cell counts across patients but also was prognostic of CD4+ T cell recovery in individuals on long-term cART. Moreover, patients with higher X4-tropic latent HIV-1 ratios in pTFH cells showed greater risks of opportunistic coinfections. These findings reveal the characteristics of latent HIV-1 in TFH cells and suggest that the ratio of X4-tropic latent HIV-1 in pTFH cells is a valuable indicator for disease progression and cART efficacy.IMPORTANCE TFH cells have been shown to harbor a significant amount of latent HIV-1; however, the viral characteristics of this reservoir and its clinical relevance remain largely unknown. In this study, we demonstrate that X4-tropic latent HIV-1 is preferentially enriched in pTFH cells, which also accurately reflects the viral tropism shift. The ratio of X4-tropic proviruses in pTFH cells but not in other memory CD4+ T cell subsets is inversely and closely correlated with blood CD4+ T cell counts and CD4+ T cell recovery rates with cART. Our data suggest that the ratio of X4-tropic provirus in peripheral TFH cells can be easily measured and reflects disease progression and treatment outcomes during cART.

[Overview of studies on DOU Han-qing's academic thoughts about acupuncture-moxibustion].

DOU Han-qing (1196-1280 A．D.) is a famous acupuncturist of the Jin and Yuan dynasties in ancient China. His academic thoughts about acupuncture and moxibustion have a higher clinical practical and theoretical value and received extensive attention in China. In the present paper, we make systematic analysis and summary about the current researches on Master DOU's academic thoughts after retrieval and screening the related research literature (published from the foundation of the People's Republic of China till now) by using electronic databases. The results showed that our understanding about Master DOU's academic thoughts are mainly based on Zhenjing Zhinan (《》，A Guide to the Classic of Acupuncture), especially the Biaoyou Fu (《》，Lyrics of Recondite Principles) and the eight influential acupoints of meridians, but less attention has been paid to books DOU Taishi Zhenjing (《》，Master DOU's Acupuncture Classics), Zhenjiu Jicheng (《》，Compendium of Acupuncture-moxibustion) and Panshijin Zhici Michuan (《》，Panshijin Esoteric Techniques on Perpendicular Insertion of Acupuncture Needles). The systematicness and depth of research on Master DOU's academic theory need being improved. We propose that we should make a comprehensive study about Master DOU's books and his life stories under the academic development history of overall background of Chinese acupuncture and moxibustion, so as to get a clear, complete and objective understanding on DOU's academic thoughts.

[Semen quality and HIV RNA level in HIV/AIDS male patients after treated by highly active antiretroviral therapy].

OBJECTIVE: To evaluate the semen quality of the HIV/AIDS male patients after treated by the highly active antiretroviral therapy (HAART) and their potential of transmitting HIV/AIDS and provide some evidence for this cohort of males who wish for parenthood. METHODS: We collected semen samples from 20 HIV/AIDS male patients who had been treated by HAART for over 6 months and wished for parenthood. We examined sperm concentration, viability and total motility and the percentage of morphologically normal sperm (MNS) using the computer-assisted semen analysis system, measured the HIV-1 RNA loads in the semen by the Cobas Amplicor Monitor test, and counted CD4+ T cells in the peripheral blood by flow cytometry. RESULTS: The patients were aged 25－40 (30.7 ± 5.05) years. After treated by HAART for 6－26 (14.24 ± 12.26) months, the count of blood CD4+ T cells was significantly increased (341－1 058 ï¼»535.76 ± 212.021ï¼½ /µl) in comparison with the baseline (226－965 ï¼»422.38 ± 200.86ï¼½ /µl). Compared with the normal value, the semen volume was increased except in 1 case (≥2 ml) while total sperm motility was decreased in 13 cases (≥40%), and so were sperm concentration in 2 cases (≥15 × 106 / ml), sperm viability in 5 (58%), the percentage of progressively motile sperm in 18 (≥32%), and the percentage of MNS in 6 (≤4%). HIV-1 RNA in the peripheral blood was <20 copies/mL in all the cases and that in the seminal plasma was also <20 copies/ml in 18 cases but >20 copies/mL in the other 2 (ï¼»4.70 × 101ï¼½ and ï¼»2.2 × 102ï¼½ copies/ml, respectively). Of the 4 couples that had sex without protective measures for over 6 months, all the 4 female partners exhibited negative HIV antibodies in regular follow-up examinations and 1 achieved spontaneous pregnancy and healthy birth, with negative HIV-1 RNA in both the mother and the baby. CONCLUSIONS: The HIV RNA level is higher in the semen than in the blood of the HIV/AIDS male patients after HAART, which indicates the potential risk of their semen transmitting HIV/AIDS to their female partners. Their sperm concentration and total sperm motility are lower than the normal value, which suggests a decreased fertility.

Anesthetic agent etiomidate induces apoptosis in N2a brain tumor cell line.

The present study identified the cytotoxic effects of etomidate on the N2a neuroblastoma cell line. Etomidate induced apoptosis in N2a cells in a concentration­dependent manner, which was confirmed by western blotting and flow cytometry. Phase contrast microscopy was used to analyze the effect of etomidate on morphological characteristics. The number of the apoptotic cells was increased and confirmed by DAPI and PI staining, which served as a characteristic hallmark of apoptosis. Additionally, etomidate led to loss of mitochondrial membrane potential and resulted in the generation of reactive oxygen species in N2a cells. The western blot analysis revealed that N2a cells treated with etomidate had a significant modulation of pro­apoptotic proteins, includingpoly ADP­ribose polymerase (PARP), cleaved PARP, caspase­9 and procaspase­3. In conclusion, the present study determined that etomidate induced cytotoxic and apoptotic effects in N2a brain tumor cells in vitro.

[The relationship between acupuncture-moxibustion theory of DOU Hanqing and Neo- Confucianism].

For the construction factors of acupuncture-moxibustion theory of DOU Hanqing, a doctor in Jin-Yuan Dynasty, we studied his life, acupuncture-moxibustion theory and culture background in the History of Medicine and Philosophy of Science & Technology. It was found that the major source of DOU Hanqing' s Confucianism was the CHENG-ZHU Neo-Confucianism. The data show that Neo-Confucianism played an important role in the enrichment of his human nature, spirit, moral cultivation and political talent, making him a scholar in the Jin-Yuan dynasty, also shaping his medical ethics, medical skill and acupuncture-moxibustion theory. The inheritance, development and application of CHENG-ZHU Neo-Confucianism provided the most fundamental thinking, research methods and patterns for DOU Hanqing to interpret the acupuncture classics, summarize his teachers' experience and develop his theoretical system. DOU Hanqing' s acupuncture-moxibustion practice and theory reached a new level, because he took acupuncture-moxibustion as one of the knowledge investigation and attainment of CHENG-ZHU Neo-Confucianism.

[Study on the origin of Dou Han-qing's academic thought of acupuncture and moxibustion].

The origin of Dou Han-qing's academic thought of acupuncture and moxibustion is studied by analysis on relative contents in the books Weisheng Baojian, Jisheng Bacui and Zhenjiu Sishu Zhenjing Zhinan and the two papers "On reinforcing-reducing manipulation" and "Questions and answers of qi and blood", holding that DOU Han-qing's academic thought of acupuncture and moxibustion is influenced more by Suwen (Plain Questions) and Nanjing (Classic on Medical Problems), not by Lingshu (Miraculous Pivot).

[Textual research and differentiation of Dou Han-qing's works catalogue].

This paper aims at probing into evolving course of DOU Han-qing's works catalogue. On the basis of summarizing and referring to study achievements of our predecessors, through analysis of book lists and relative works and chapters, it is hold that the catalogue which were not attained by ZHULiang-neng possibly are the contents of acupuncture reinforcing and reducing methods; the books printed and published by ZHULiang-neng include the contents of both channels and acupoints; the book, Zhinan, which was attained by DOUGui-fang, includes the content catalogue of needling methods; Fu Zhenjiu Zashuo in Zhenjiu Sisu. Zhenjiu Zhinan also were extracted by DOU Gui-fang from Illustrated Manual of Acupoints of the Bronze Figure, and The Zhenjiu Biji Taiyi Zhi Tuxu and Dongzhi Yezhe Gongshuo should belong to The Fu Zhenjiu Zashuo.

Induction of specific cytolytic T lymphocytes using fusions of hepatocellular carcinoma (HCC) patient-derived dendritic cells and allogeneic HCC cell line.

BACKGROUND/AIMS: To assess the ability of hepatocellular carcinoma (HCC) patient-derived dendritic cells (DCs) fused with allogeneic HCC cell line to activate autologous lymphocytes to generate specific cytotoxic T lymphocytes (CTL) in vitro. METHODOLOGY: DCs were obtained by culturing adherent peripheral blood mononuclear cells (PBMC) from HCC patients in the presence of 100 microg/L recombinant human granulocyte/ macrophage- colony stimulating factor (rhGM-CSF) and 20 microg/L interleukin-4 (rhIL-4) for 1 week in vitro. DCs were fused with allogeneic HCC cell line HepG2 cells using polythyleneglycol (PEG), and the fusion cells were designated as DCs/HepG2. By labeling DCs and HepG2 with green and red fluoresceins, respectively, the cellular fusion was examined under fluorescence microscope. The ability of DCs/HepG2 to stimulate proliferation and differentiation of autologous lymphocytes was assessed by MTT method, and the specific killing efficacy of DCs/HepG2-induced CTL against HepG2 was evaluated. RESULTS: HCC patient-derived DCs expressed a certain level of CD1a, HLA-DR, CD54, CD80 and CD86. Fluorescence microscopic examination demonstrated that co-incubation of DCs and HepG2 in the presence of PEG lead to generation of DCs/HepG2. In the mixed lymphocyte reaction assay, DCs/HepG2 had a significantly greater ability to activate proliferation of autologous lymphocytes, as compared with DCs alone, DCs plus HepG2, HepG2 alone and medium control (P<0.05). The DCs/HepG2-activated CTL showed a potent specific killing efficacy against HepG2 cells. CONCLUSIONS: Fusions of HCC patient-derived DCs and allogeneic HCC cell line could efficiently stimulate autologous lymphocytes to generate tumor-specific CTL in vitro. It might represent a promising approach of immunotherapy for HCC.