内疫病机概念的提出与治则探讨

吴有性的"杂气论"和中医的"天人合一"哲学观点是提出内疫病机概念的文献依据;现代医学中与肿瘤发病相关基因的发现、与自身免疫病相关的自身抗体的发现,以及这两类疾病的发病机制是提出内疫病机概念的临床依据。内疫病机认为,人体自身有杂气,健康人体杂气受正气制约,正气一旦亏虚,则可能对杂气失于控制,杂气变逆,甚而作疫,导致疾病。针对内疫病机提出促邪归正治疗原则,即扶正与攻毒并施,促使内生的疫气重新归顺正气,为正气制约,与诱导疗法暗合。

从内疫谈抗中性粒细胞胞浆抗体相关性血管炎的发病与治疗

目的:从内疫探讨抗中性粒细胞胞浆抗体(ANCA)相关性血管炎(AASV)的发病与治疗。方法:从人体杂气与内疫、人体杂气与AASV发病以及人体杂气与AASV治疗的相关性出发,对ANCA相关性血管炎的发病与治疗进行阐述。结果:正气亏虚是导致内疫病机的直接病因,可将AASV的发生归咎于人体自身杂气的失制。治疗采用扶正与攻毒联合用药,攻补并施,形成促使变逆之气向正气转化的合力,实现扶正与攻毒两种治法在针对病机上的矛盾统一。结论:通过对AASV的内疫病机讨论,为继承及使用杂气病因学说治疗AASV提供了可能,以期为AASV的中医治疗提供新的思路。

从内生伏毒谈内疫病机与治法

伏毒学说认为:内生伏毒是导致诸多内科疑难病的重要病因;内疫病机是根据杂气病因学提出的假说,这两个理论均源于中医温病学。针对内生伏毒的辨证原则和治法方药可以用于内疫病机导致疾病,以实现促邪归正治疗宗旨。通过对内疫病机的认识可以加深对伏毒学说的理解,有利于掌握伏毒的辨证原则和治法方药,而伏毒学说则为内疫病机提供了从理论假说走向临床实践的桥梁。

从杂气病因学谈恶性肿瘤的发病与治疗

杂气病因学说是温病学阐释疫邪发病的重要基础理论。笔者认为，人体同自然界一样具有杂气，并藉此提出“内疫”的概念。通过对恶性肿瘤的现代医学发病机制分析，发现其符合内疫病机特点，现就此阐述如下，与同道探讨，以期为恶性肿瘤的中医治疗提供新的思路。

BIO-INSPIRED SELF-ADAPTIVE MANUFACTURING SYSTEM CONTROL ARCHITECTURE

Future manufacturing systems need to cope with frequent changes and disturbances, therefore their control architectures require constant adaptability, agility, stability, self-organization, intelligence, and robustness. Bio-inspired manufacturing system can well satisfy these requirements. For this purpose, by referencing the biological organization structure and the mechanism, a bio-inspired manufacturing cell is presented from a novel view, and then a bio-inspired self-adaptive manufacturing model is established based on the ultra-short feedback mechanism of the neuro-endocrine system. A hio-inspired self-adaptive manufacturing system coordinated model is also established based on the neuro-endocrine-immunity system （NEIS）. Finally, an example based on pheromone communication mechanism indicates that the robustness of the whole manufacturing system is improved by bio-inspired technologies.

Clinico-pathologic significance of neuroendocrine cells in gastric carcer tissue

AIMS To elucidate the biological and clinicopathological signifi- cance of neuroendocrine(NE)cells in gastric carcinoma(GC). METHODS One hundred and eighty-nine cases of various histo- logical types of GC were observed using light microscopy, histochemistry,immunohistochemistry and electron microscopy. Of them 127 patients were followed up. RESULTS Chromogranin A(CgA)positive GC was demonstrat- ed in 85 cases(45.0%).The typos of NE cells in GC were probed using 9 kinds of hormone antibodies 49 cases(67.2%) contained more than one hormone.NE cells were found more often in poorly differentiated GC than in well differentiated ones(P<0. 01).Expression of some kinds of hormone Was relative to the dif- ferentiation and histological types of GC.BOM,CT(P<0.01) GAST and 5-HT(P<0.05)were more expressed in poorly differ- entiated cases than in well defferentiated ones.Nineteen cases of the matastatic foci in regional lymph nodes were found to have CgA positive cancer cells.The presence of HCG in matastatic lymph nodes was more often observed than that of other hormone (P<0.01).The survival rate of patients with NE positive GC was 38.9% and negative GC 52.7%.Five of 7 patients(71. 4%)with somatostatin positive GC still survived at follow up of 33-66 months,but 4 patients with HCG positive GC died in 12-29 months. CONCLUSIONS The NE cells occur more frequently in poorly differentiated GC.Certain hormones appear to be related to the metastasis and prognosis.

Alterations of intestinal immune function and regulatory effects of L-arginine in experimental severe acute pancreatitis rats

AIM： To discuss the changes of intestinal mucosal immune function in rats with experimental severe acute pancreatitis （SAP） and the regulatory effect of L-arginine. METHODS： Male adult Wistar rats were randomly divided into pancreatitis group, sham-operation group, and L-arginine treatment group. Animals were killed at 24, 48, and 72 h after SAP models were developed and specimens were harvested. Endotoxin concentration in portal vein was determined by limulus endotoxin analysis kit. CD3＋, CD4＋, CD8＋ T lymphoo/tes in intestinal mucosal lamina propria were examined by immunohistochemistry. Secretory immunoglobulin A （SIgA） in cecum feces was examined by radioimmunoassay. RESULTS： Compared to the control group, plasma endotoxin concentration in the portal vein increased, percentage of CD3＋ and CD4＋ T lymphocyte subsets in the end of intestinal mucosal lamina propria reduced significantly, CD4＋/CD8＋ ratio decreased, and SIgA concentrations in cecum feces reduced at 24, 48, and 72 h after SAP developed. Compared to SAP group, the L-arginine treatment group had a lower level of plasma endotoxin concentration in the portal vein, a higher CD3＋ and CD4＋ T lymphocyte percentage in the end of intestinal mucosal lamina propria, an increased ratio of CD4＋/CD8＋ and a higher SIgA concentration in cecum feces. CONCLUSION： Intestinal immune suppression occurs in the early stage of SAP rats, which may be the main reason for bacterial and endotoxin translocation. L-arginine can improve the intestinal immunity and reduce bacterial and endotoxin translocation in SAP rats.

Intraductal neoplasm of the intrahepatic bile duct: Clinicopathological study of 24 cases

AIM: To investigate the clinicopathological features of intraductal neoplasm of the intrahepatic bile duct (INihB). METHODS: Clinicopathological features of 24 cases of INihB, which were previously diagnosed as biliary papillomatosis or intraductal growth of intrahepatic biliary neoplasm, were reviewed. Mucin immunohistochemistry was performed for mucin (MUC)1, MUC2, MUC5AC and MUC6. Ki-67, P53 and β-catenin immunoreactivity were also examined. We categorized each tumor as adenoma (low grade), borderline (intermediate grade), and malignant (carcinoma in situ , high grade including tumors with microinvasion). RESULTS: Among 24 cases of INihB, we identified 24 tumors. Twenty of 24 tumors (83%) were composed of a papillary structure; the same feature observed in intraductal papillary neoplasm of the bile duct (IPNB). In contrast, the remaining four tumors (17%) showed both tubular and papillary structures. In three of the four tumors (75%), macroscopic mucin secretion was limited but microscopic intracellular mucin was evident. Histologically, 16 tumors (67%) were malignant, three (12%) were borderline, and five (21%) were adenoma. Microinvasion was found in four cases (17%). Immunohistochemical analysis revealed that MUC1 was not expressed in the borderline/adenoma group but was expressed only in malignant lesions (P = 0.0095). Ki-67 labeling index (LI) was significantly higher in the malignant group than in the borderline/adenoma group (22.2 ± 15.5 vs 7.5 ± 6.3, P < 0.01). In the 16 malignant cases, expression of MUC5AC showed borderline significant association with high Ki-67 LI (P = 0.0622). Nuclear expression of β-catenin was observed in two (8%) of the 24 tumors, and these two tumors also showed MUC1 expression. P53 was negative in all tumors. CONCLUSION: Some cases of INihB have a tubular structure, and are subcategorized as IPNB with tubular structure. MUC1 expression in INihB correlates positively with degree of malignancy.

Immunohistochemical study on distribution of endocrine cells in gastrointestinal tract of flower fish (Pseudophoxinus antalyae)

AIM： To detect distribution and relative frequency of endocrine cells in gastrointestinal tract of flower fish （Pseudophoxinus antalyae）. METHODS： The intestinal tract of flower fish was divided into four portions from proximal to distal; the enlarged area after oesophagus and anterior, middle and posterior intestine. Immunohistochemical method using the peroxidase anti-peroxidase complex was employed. All antisera between four portions of flower fish were compared using ANOVA. RESULTS： Eleven types of gut endocrine cells were determined; they were immunoreactive for calcitonin gene related peptide, substance P, vasoactive intestinal peptide, bombesin, somatostatin-14, secretin, TrkA, TrkB, TrkC, neurotensin, neuropeptide Y, which were found in almost all portions of the gastrointestinal tract. CONCLUSION： The regional frequency of immunoreactive distribution and relative cells in the flower fish, Pseudophoxinus antalyae, are essentially similar to those of other fish.

The inhibition of lung cancer cell growth by intracellular immunization with LC-1 ScFv

A monoclonal antibody, LC-1, recognizing lung cancer associated common antigens was obtained in authors’ laboratory. Its single chain Fv fragment (ScFv) named LC-1 ScFv was constructed based on recombinant phage displayed techniques. For expression on cell membrane, LC-1 ScFv was cloned into pDisplay vector, which directed the cloned gene to express as cell membrane bound protein. The resulting plasmid was sequenced and then introduced by the lipofectin method into a lung adenocarcinoma cell line SPC-A-1. G418 resistant cells were obtained by G418 selection. After transfection, LC-1 ScFv expression was observed by Western blot analysis and the expression of cognate antigens was down-regulated as shown in ELISA assay. SPC-A-1-pDisplay-ScFv cells grew in vitro at lower speed than the control intact cells and the cells transfected with vacant vector. Flow cytometry analysis detected a substantial increase in G1 phase and decrease in S phase in population of SPC-A-1-pDisplay-ScFv cells compared to SPC-A -1 and SPC-A1-pDisplay cells. Semi-quantitative RT-PCR analysis showed that c-myc expression was down-regulated in SPC-A-1-pDisplay-ScFv cells. It seems that the antigens recognized by LC-1 may be in some way involved in a growth stimulating pathway and the antibody blocking of the function of the antigens shut down the pathway and thus down-regulate the expression of c-myc and growth of the cells.

Innate immunity in inflammatory bowel disease

The human intestinal tract is home to an enormous bacterial flora. The host defense against microorganisms can be divided into innate and adaptive immunity. The former is the most immediate line of response to immunologic challenges presented by bacteria, viruses, and fungi. The mucosal immune system has evolved to balance the need to respond to pathogens while co-existing with commensal bacteria and food antigens. In inflammatory bowel disease （IBD）, this hyporesponsiveness or tolerance breaks down and inflammation supervenes driven by the intestinal microbial flora. Bacteria contain compounds and are recognized by a variety of receptors, including Toll-like receptors （TLRs） and NODs （a family of intracellular bacterial sensors） and are potent stimuli of innate immune responses. Several mutations in these receptors have been associated with development of IBD.

Diagnosis of autoimmune gastritis by high resolution magnification endoscopy

Endoscopic visualisation of gastric atrophy is usually not feasible with conven.tional endoscopy. Magnifying endoscopy is helpful to analyze the subepithelial microvascular architecture as well as the mucosal surface microstructure without tissue biopsy. Using this technique we were able to describe the normal gastric microvasculature pattern and we also identified characteristic patterns in two cases of autoimmune atrophic gastritis.

Expressions of claudin-4 and claudin-1 in endometrial cancer and their significance

Objective:To observe the expressions of claudin-4 and claudin-1 in endometrial cancer and explore their correlations with clinicopathological parameters of endometrial cancer.Methods:Immunohistochemical methods(SP) were used to detect the expressions of claudin-4 and claudin-1 in 52 tissue samples of endometrial cancer,24 of atypical hyperplasia,20 of pericancerous endometrium,and 19 of endometrium at proliferative phase.And then the expressions were analyzed statistically to find out the correlations with clinicopathological parameters of endometrial cancer.Results:Positive rate of claudin-4 was 36.8%,70.8% and 90.4% in endometrium at proliferative phase,atypical hyperplasia and endometrial cancer,respectively,with significantly differences between them(P<0.05),and it was statistically different between pericancer endometrium and endometrial cancer(P<0.05).Positive rate of claudin-1 was 89.5%,66.7% and 63.5%,respectively showing a descending tendency and significantly differences between endometrium at proliferative phase and endometrial caner(P<0.05),and it was also statistically significantly different between pericancer endometrium and endometrial cancer(P<0.05).The high expression rate of claudin-4 was related to invasion depth,but not to histological grading,pathological staging or lymph node metastasis of endometrial cancer,and the low expression of claudin-1 in endometrial cancer was not associated with histological grading,pathological staging,invasion depth or lymph node metastasis.Conclusion:The expression levels of claudin-4 and claudin-1 are correlated with onset and development of endometrial cancer.

PROTECTION AGAINST LEPTOSPIROSIS BY IMMU NIZATION WITH PLASMID DNA ENCODING 33 kDa ENDOFLAGELLIN OF L.INTERROGANS SEROVAR LAI

To evaluate how the efficacy of DNA inocutation affects the ability to raise protective immunity against Leptospira. [WT5”BX]Methods.[WT5”BZ] A pair of oligonucleotide primers were designed to amplify the endoflagellar gene of L. interrogans sensu stricto serovar lai. An approximately 840bp fragment was generated with PCR and inserted into VR1012, a plasmid DNA expression vector, after the fragment and VR1012 were digested respectively with EcoRV and Sal I. A recombinant plasmid designated as VR1012+flaB2 was obtained. The vector, VR1012 consits of a pUC18 backbone with the cytomegalovirus(CMV) IE1 enhancer, promoter, and intron A, transcription regulatory elements and the BGH polyadenylation sequences driving the expressing of leptospiral endoflagellar gene of L. interrogans sensu stricto serovar lai. Plasmid encoding leptospiral endoflagellin gene was injected into quadriceps of NZW rabbits. [WT5”BX]Results.[WT5”BZ]This resulted in the generation of specific leptospiral antibody with high ELISA titer (1:32768) in the rabbits. Immuno/protection was performed in guinea pigs without adjuvant. The group“VR1012+flaB2” showed higher survival rate(90%,9/10 animals),compared with the group “VR1012 lack flaB2” and the group “normal saline”. [WT5”BX]Conclusion.[WT5”BZ]The technique of DNA vaccine has potential advantages over certain other vaccine preparation technologies. However whether DNA vaccine will be useful for vaccine development remains to be tested.

Esophageal mesenchymal tumors:Endoscopy,pathology and immunohistochemistry

AIM： To study the endoscopic, pathological and immunoo histochemical features of esophageal mesenchymal tumors. METHODS： Twenty-nine patients diagnosed as esophageal rnysenchymal tumors by electronic endoscopy and endoscopic ultrasound （EUS） were observed under light microscopes, and all tissues were stained by the immunohistochemical method. The expression of CD117, CD34, SMA and desmin were measured by staining intensity of cells and positive cell ratios. RESULTS： Endoscopically, esophageal gastrointestinal stromal tumors （GISTs） and leiomyomas （LMs） had similar appearances, showing submucosal protuberant lesions. They all showed low echo images originated from the muscularis propria or muscularis mucosa on EUS. Endoscopy and EUS could not exactly differentiate esophageal GISTs from LMs. Microscopically, there were two kinds of cells： spindle cell type and epitheloid cell type in esophageal GISTs. Leiomyomas and leiomyosarcornas were only of spindle cell type. One malignancy was found in five cases of esophageal GISTs, and one malignancy in 24 cases of leiomyomas and leiomyosarcomas. Using Fisher＇s exact method, the differences of malignant lesion proportion were not significant between esophageal LMs and GISTs, 1/5 vs 1/24 （P 〉 0.05）. All cases of esophageal GISTs were positive for CD117, and 3 cases were also positive for CD34. The 24 cases of leiomyomas and leiomyosarcomas were all negative for CD117 and CD34. The differences of positive rates of CD117 and CD34 were significant between esophageal GISTs and LMs, 5/5 vs 0/24, 3/5 vs 0/24 （P 〈 0.005）. All leiomyomas and leiomyosarcomas were positive for SMA, and desmin. Among 5 cases of esophageal GISTs, 2 cases were SMA positive, and 1 case was desmin positive. The differences in positive rates and expression intensity of SMA and desmin were significant between esophageal LMs and GISTs, 24/24 vs 2/5, 24/24 vs 115 （P 〈 0.005）. CONCLUSION： The most common esophageal mesenchymal tumors are leiomyomas, and esophageal GISTs are less common. Most of esophageal LMs and GISTs are benign. Endoscopy and EUS are the effective methods to diagnose esophageal mesenchymal tumors and they can provide useful information for the treatment of these tumors. However, they cannot exactly differentiate esophageal GISTs from LMs. Pathological, especially immunohistochemical features are useful to differentiate GISTs from leiomyomas.

1,25-dihydroxyvitamin D_3 regulates LPS-induced cytokine production and reduces mortality in rats

AIM： To study the immunoregulatory effect of 1,25-dihydroxyvitamin-D3 Von dominant Thl response in rats. METHODS： Sixty adult Lewis rats were randomized into three groups. Rats in group 1 （n=25） were treated with 1,25-（OH）2D3 first and then challenged with LPS, rats in group 2 （n=25） were treated with vehicle first and then challenged with LPS. Ten animals in groups 1 and 2 were preserved for mortality observation. The remaining animals were injected （i.p） with endotoxin, 24 h after the last administration of 1,25-（OH）2D3 and vehicle. Rats in group 3 （n=10） were treated with 1,25-（OH）2D3 only. Serum IL-12, IFN-y, IL-2 and IL-4 levels were measured and target gene of 1,25-（OH）2D3 on Th cells was studied after 6 h. Gene abundance was verified by real-time quantitative PCR. RESULTS： No death occurred in rats pretreated with 1,25-（OH）2D3 after LPS injection. Death occurred 9 h after LPS injection in rats pretreated with the vehicle, and the number of deaths was 5 within 24 h, with a mortality rate of 50%. There was no change in the number of deaths within 96 h. Six hours after endotoxin stimulation, serum IL-12 and IFN-y levels decreased significantly in rats pretreated with 1,25-（OH）2D3 as compared with those in rats pretreated with the vehicle. The serum content of these two cytokines was very low in rats not challenged by endotoxin, and there was a significant difference as compared with the previous two groups. CONCLUSION： 1,25-（OH）2D3 attenuates injuryinduced by the lethal dose of 1PS, regulates Thl and Th2 cells at the transcription level, and dominantly responds to cytokine production in rats.

COMMENTS ON THE HOLISTIC ACTION OF ACUPUNCTURE IN VIEW OF ITS INFLUENCE ON NERVE-ENDOCRINE-IMMUNE NETWORK

In the present paper, the author makes some comments on the holistic action of acupuncture from the regulative effect on the nerve-endocrine-immune network, and the close relationship between the nerve-endocrine-immune network and the meridian-collateral system of TCM. The wholism concept of TCM refers to the organism being an integrated entirety, and the regulatory effect of acupuncture on functional activities of the organism relies on the integral connection of the acupoint-meridian-collateral-zangfu-organ system. When stimulated with acupuncture, the human body will brings its potential force into full play in preventing and treating diseases.

Changes in intestinal mucosal immune barrier in rats with endotoxemia

AIM： To investigate the dysfunction of the immunological barrier of the intestinal mucosa during endotoxemia and to elucidate the potential mechanism of this dysfunction. METHODS： Male Wistar rats were randomly distributed into two groups： control group and lipopolysaccharide （LPS） group. Endotoxemia was induced by a single caudal venous injection of LPS. Animals were sacrificed in batches 2, 6, 12 and 24 h after LPS infusion. The number of microfold （M）-cells, dendritic cells （DCs）, CD4＋ T cells, CD8＋ T cells, regulatory T （Tr） cells and IgA＋ B cells in the intestinal mucosa were counted after immunohistochemical staining. Apoptotic lymphocytes were counted after TUNEL staining. The levels of interleukin （IL）-4, interferon （IFN）-γ, and forkhead box P3 （Foxp3） in mucosal homogenates were measured by ELISA. The secretory IgA （sIgA） content in the total protein of one milligram of small intestinal mucus was detected using a radioimmunological assay.RESULTS： This research demonstrated that LPS-induced endotoxemia results in small intestinal mucosa injury. The number of M-cells, DCs, CD8~ T cells, and IgA~ B cells were decreased while Tr cell and apoptotic lymphocyte numbers were increased significantly. The number of CD4＋ T cells increased in the early stages and then slightly decreased by 24 h. The level of IL-4 significantly increased in the early stages and then reversed by the end of the study period. The level of IFN-T increased slightly in the early stages and then decreased markedly by the 24 h time point. Level of Foxp3 increased whereas sIgA level decreased.CONCLUSION： Mucosal immune dysfunction forms part of the intestinal barrier injury during endotoxemia. The increased number and function of Tr cells as well as lymphocyte apoptosis result in mucosal immunode- ficiency.

Endoscopic therapy for gastric stromal tumors originating from the muscularis propria

AIM:To explore endoscopic therapy methods for gastric stromal tumors originating from the muscularis propria.METHODS:For 69 cases diagnosed as gastric stromal tumors originating from the muscularis propria,three types of endoscopic therapy were selected,based on the size of the tumor.These methods included endoscopic ligation and resection(ELR),endoscopic submucosal excavation(ESE) and endoscopic full-thickness resection(EFR).The wound surface and the perforation of the gastric wall were closed with metal clips.Immunohistostaining for CD34,CD117,Dog-1,S-100 and smooth muscle actin(SMA) was performed on the resected tumors.RESULTS:A total of 38 cases in which the tumor size was less than 1.2 cm were treated with ELR;three cases were complicated by perforation,and the perforations were closed with metal clips.Additionally,18 cases in which the tumor size was more than 1.5 cm were treated with ESE,and no perforation occurred.Finally,13 cases in which the tumor size was more than 2.0 cm were treated with EFR;all of the cases were complicated by artificial perforation,and all of the perforations were closed with metal clips.All of the 69 cases recovered with medical treatment,and none required surgical operation.Immunohistostaining demonstrated that among all of the 69 gastric stromal tumors diagnosed by gastroscopy,12 cases were gastric leiomyomas(SMA-positive),and the other 57 cases were gastric stromal tumors.CONCLUSION:Gastric stromal tumors originating from the muscularis propria can be treated successfully with endoscopic techniques,which could replace certain surgical operations and should be considered for further application.

A low prevalence of H pylori and endoscopic findings in HIV-positive Chinese patients with gastrointestinal symptoms

AIM： To compare the prevalence of H pylori infection, peptic ulcer, cytomegalovirus （CNV） infection and Candida esophagitis in human immunodeficiency virus （HIV）- positive and HIV-negative patients, and evaluate the impact of CD4 lymphocyte on H pylori and opportunistic infections. METHODS： A total of 151 patients （122 HIV-positive and 29 HIV-negative） with gastrointestinal symptoms were examined by upper endoscopy and biopsy. Samples were assessed to determine the prevalence of Hpylori infection, CMV, candida esophagitis and histologic chronic gastritis. RESULTS： The prevalence of Hpylori was less common in HIV-positive patients （22.1%） than in HIV-negative controls （44.8%; P 〈 0.05）, and the prevalence of H pylori displayed a direct correlation with CD4 count stratification in HIV-positive patients. In comparison with HIV-negative group, HIV-positive patients had a lower incidence of peptic ulcer （20.7% vs 4.1%; P 〈 0.01）, but a higher prevalence of chronic atrophy gastritis （6.9% vs 24.6%; P 〈 0.05）, Candida esophagitis and CMV infection. Unlike HIV-negative group, H pylori infection had a close relationship to chronic active gastritis （P 〈 0.05）. In HIV-positive patients, chronic active gastritis was not significantly different between those with Hpylori infection and those without. CONCLUSION： The lower prevalence of H pylori infection and peptic ulcer in HIV-positive patients with gastrointestinal symptoms suggests a different mechanism of peptic ulcerogenesis and a different role of H pylori infection in chronic active gastritis and peptic ulcer. The pathogen of chronic active gastritis in HIV-positive patients may be different from the general population that is closely related to Hpylori infection.