Ultrastructure Characteristics of Different Chinese Medicine Syndromes of Helicobacter pylori-Correlated Gastric Diseases.

OBJECTIVE: To explore the ultrastructure characteristics of patients with dampness-heat of Pi (Spleen)-Wei (Stomach) syndrome (DHPW) and Pi-qi deficiency syndrome (PQD), both of which are Helicobacter pylori (Hp)-correlated gastric diseases (HPCG), and implicate a helpful hint for the clinical microcosmic syndrome differentiation. METHODS: Fourteen gastric mucosa samples from 6 chronic gastritis (CG) and 6 active peptic ulcer (including 8 DHPW, 4 PQD) as well as 2 healthy volunteers were collected and tested for Hp infection. The ultrastructure of gastric mucosa was observed under the transmission electron microscope (TEM). RESULTS: Among 14 gastric mucosa samples, 8 of them were Hp positive (6 DHPW and 2 PQD), which were all accordance with the results screened by supermicro-pathological method. Under TEM, the normal gastric mucosa, with tidy microvilli and abundant in mucus granules, mitochondria and rough endoplasmic reticulum distributed evenly, and with smooth nucleus membrane. But in those specimens of DHPW with Hp infection, microvilli were presented with burr shape. Especially, those samples from dampness-heat syndrome with predominant heat type (DHSH) patients were more obvious, with microvilli damaged, mitochondria concentrated and distributed in disorder, secretory tubule extended. In dampness-heat syndrome with predominant dampness type (DHSD) patients, mucus granules aggregated obviously, mitochondria swelled and blurred, and rough endoplasmic reticulum crowded. For 2 samples of DHPW without Hp infection, their microvilli were intact, with mitochondria increased and gathered but well-distributed, and secretory tubule extended mildly. In 2 PQD patients with Hp positive, the specimens of microvilli were sparse, and their mucus granules and mitochondria were decreased, with fractured crests and vacuole, secretory tubules extension to nucleus membrane, and rough endoplasmic reticulum extension in a pool-like way, and nucleus condensed. The 2 samples from PQD patients without Hp infection were characterized with intact microvilli, decreased mitochondria, fractured crest and extended rough endoplasmic reticulum in a pool-like way. CONCLUSION: It's obviously different in ultrastructure of DHPW and PQD patients under TEM, which may give a helpful hint for the microcosmic syndrome differentiation of HPCG.

[Expressions of MMP-7 and TIMP-1 in Chronic Stomach Disease Patients with Different Syndromes of Chinese Medicine].

Objective To observe the expressions of matrix metalloproteinase-7 (MMP-7) and tissue inhibitor of metalloproteinase-1 (TIMP-1 ) in chronic stomach disease patients with different syn- dromes of Chinese medicine (CM) , and their relationships with Helicobacter pylori ( Hp ) infection. Meth- ods Totally 117 chronic stomach disease patients were recruited, and 11 healthy volunteers were also recruited. Chronic stomach disease patients were assigned to Pi-Wei dampness-heat syndrome (PWDHS, 57 cases) , disharmony of Gan and Wei syndrome (DGWS, 30 cases) , and Pi qi deficiency syndrome (PQDS, 30 cases) by syndrome typing. Healthy volunteers were recruited as the healthy con- trol group. Hp infection was detected using methylene blue dyeing and rapid urease test (RUT). The degree of inflammation was observed by conventional HE staining. The protein expressions of MMP-7 and TIMP-1 were detected qualitatively and positioningly using immunohistochemical method. Results Patients with PWDHS and patients with DGWS had equivalent Hp infection rate and degree. They showed a slightly increasing tendency than patients with PQDS, but with no statistical difference (P >0. 05). Com- pared with PWDHS and PQDS groups, more severe inflammation of mucosa occurred in patients with DG- WS (P <0. 05). More severe inflammation of mucosa occurred in patients with PWDHS than in those with PQDS, but with no statistical difference (P >0. 05). The severity of gastric mucosal inflammatory activity was sequenced from high to low as PWDHS, DGWS, PQDS, all with statistical difference (P <0. 05). Compared with Hp negative patients, the gastric mucosal inflammatory activity was more severe in Hp positive patients with PWDHS, DGWS, PQDS. The gastric mucosal inflammatory activity was more se- vere in Hp positive patients with PWDHS and PQDS (P <0. 05). Compared with the healthy control group, the expression level of TIMP-1 in gastric mucosa increased in patients with PWDHS, DGWS, PQDS (P <0. 05, P <0. 01) ; the expression level of MMP-7 increased in Hp negative patients with PWDHS (P < 0. 05). Compared with Hp negative patients with PWDHS, the expression level of MMP-7 decreased in Hp positive patients with PWDHS (P <0. 05). Compared with the PQDS group, the expression level of TIMP-1 decreased in the PWDHS group (P <0. 01). The severity of gastric mucosal inflammation was negatively correlated with the expression level of MMP-7, and positively correlated with the expression level of TIMP- 1 (P <0. 01). Hp infection degree was not obviously correlated with the expression level of MMP-7 in gastric mucosa (P >0. 05) , but positively correlated with the expression level of TIMP-1 in gastric mucosa (P <0. 05). Of them, the expression level of MMP-7 in gastric mucosa was positively correlated with the expression level of TIMP-1 in gastric mucosa (P <0. 01). Conclusions Comparatively lower expression of MMP-7 in gastric mucosal inflammation and imbalanced expression of TIMP-1 might be two of the pathogeneses of chronic stomach disease. Their various expressions in different CM syndromes might have certain expositions for microscopic research on "different syndromes of the same disease". Emotional fluctuation might also be one of important factors for chronic stomach disease.

[Relation between dysbacteriosis of Helicobacter pylori and Lactobacillus acidophilus and the pathogenesis of chronic gastropathy patients of Pi-Wei damp-heat syndrome].

Pi-Wei theory is an important component of the basic theory of Chinese medicine. The pathogenesis of Pi-Wei damp-heat syndrome is an important content of Pi-Wei theory. The tongue coating is one of the most important signs reflecting Pi-Wei damp-heat syndrome. From the perspective of microecology and pathogenesis, the microbial disequilibrium caused by quantity changes of Helicobacter pylori (Hp) and Lactobacillus acidophilus (LA) and their interaction in the gastric mucosa and the tongue coating might have certain correlation with "mutual struggle between the evil and the vital qi, the disequilibrium between yin and yang". The pathogenesis features of chronic gastritis patients of Pi-Wei damp-heat syndrome was initially proposed in this article.

[Relationship between Pi-Wei damp-heat syndrome with expressions of nuclear factor-kappaB mRNA and heat shock protein 70 mRNA in patients with chronic gastritis].

OBJECTIVE: To explore the relationship between the genesis of Pi-Wei damp-heat syndrome (PDS) and the Helicobacter pylori (HP) infection in patients with chronic gastritis (CG) by observing the levels of nuclear factor kappaB (NF-kappaB) mRNA and heat shock protein 70 (HSP70) expressions. METHODS: Gastric mucous membrane tissues collected through gastroscopy from gastric antrum of 51 CG patients, 36 of PDS type (CG-PDS) and 15 of Pi-qi deficiency syndrome (CG-non-PDS) type, and 8 healthy persons (as control) were examined to diagnose the inflammation level with HE stain and to detect the existence of HP infection by rapid urease test and methylthioninium chloride stain. The mRNA expressions of NF-kappaB and HSP70 in the tissues were determined quantitatively with FQ-PCR as well. RESULTS: Patients of CG-PDS showed higher level of HSP70 mRNA expression than in the control; and level of NF-kappaB mRNA expression was higher than that in the control and CG-non-PDS (all P<0.05); but both expressions were insignificantly different in CG-PDS patients with positive or negative HP infection (P>0.05). Positive correlation of the two expressions was shown in CG-PDS with negative HP infection. CONCLUSION: NF-kappaB and HSP70 may partially embody, in some extent, the power of vital qi and evil qi in the organism; the over-expressed NF-kappaB and HSP70 may indicate the severe fighting between evil qi and vital qi, and both would be influenced to a certain degree in the fighting process.

Relationship between gene expressions of aquaporin 3 and 4 and various degrees of spleen-stomach dampness-heat syndrome in chronic superficial gastritis.

OBJECTIVE: To explore the relationship between gene expressions of aquaporin (AQP) 3 and 4 and various degrees of spleen-stomach dampness-heat syndrome in chronic superficial gastritis (CSG). METHODS: Twenty-four CSG patients were divided into three groups according to the degrees of spleen-stomach dampness-heat syndrome: mild, moderate and severe groups. There were 8 patients in each group, and another 8 healthy persons were selected as normal control. Samples of mucosa of upper stomach in the included patients and normal persons were collected by gastroscopy, and then were stored in liquid nitrogen. The gene expressions of AQP3 and AQP4 in gastric tissue were examined by fluorescence quantitative polymerase chain reaction. RESULTS: The gene expressions of AQP3 and AQP4 in the moderate and severe groups were higher than that in the mild group and normal group (P<0.05, P<0.01). The gene expression of AQP3 in the severe group was higher than that in the moderate group (P<0.05). CONCLUSION: The gene expressions of AQP3 and AQP4 in gastric mucosa are different in patients with various degrees of spleen-stomach dampness-heat syndrome of CSG. There is a relationship between expressions of AQP3 and AQP4 genes and the degrees of spleen-stomach dampness-heat syndrome, and the gene expressions will increase with the aggravation of the dampness-heat syndrome.

Bioinformatics research on chronic superficial gastritis of Pi-deficiency syndrome by gene arrays.

OBJECTIVE: To determine the bioinformatical characteristics of differential gene expression in patients with chronic superficial gastritis (CSG) with the Pi-deficiency syndrome (PDS) and those of the non-Pi-deficiency syndrome (non-PDS), i.e. patients of CSG with Pi-Wei dampnese-heat syndrome and healthy persons. METHODS: With the BRB-Array Tools software package, original data collection and bioinformatic: analysis of gene arrays were conducted in 6 CSG patients of PDS (CSG-PDS), 6 CSG patients of non-PDS (CSG-nPDS), and 6 healthy volunteers (Normal). RESULTS: Compared with non-PDS, the gene expressions: in PDS with regards to protein synthesis, energy metabolism, immune reaction and ionic transport tended to be down-regulated, while those concerning secretion, cytoskeleton and ubiquitinization were up-regulated dominantly. CONCLUSIONS: The two kinds of samples, CSG-PDS/Normal and CSG-PDS/CSG-nPDS, have their respective gene expression profiles with different characteristics. Gene expression profile has certain referential significance in syndrome classification.

[The virtuous effect of heat shock protein 70 and evil effect of nuclear factor-kappa B in patients with chronic gastric disease of PI-Wie damp-heat syndrome type].

It has been proved in recent studies that the chronic gastric disease (CGD) of Pi-Wei damp-heat syndrome type (CGD-PWDH) is closely related with heat shock protein 70 (HSP70) and nuclear factor-kappa B (NF-kappaB). HSP70 can protect the auto-stability of cells and elevate the immune function in organism against tumor or multiple exogenous pathogens. Increasing of NF-kappaB expression presents in case of Helicobacter pylori (Hp) stimulation, it could induce inflammatory reaction, while inflammation factors could act inversely to enhance the expression of NF-kappaB, thus to cause and expand the damage of gastric mucosa. In addition, HSP shows blocking effect on the activation and expression of NF-kappaB. So, the author considered that in patients of Hp associated CGD-PEDH, HSP 70 exhibits the effect as that of "vital energy" and NF-kappaB play a role as the "evil qi" in Chinese medicine, the expressions of the two may embody the vital-evil combating manner of Pi-Wei damp-heat syndrome.

[Thoughts on pathogenesis of Helicobacter pylori related gastrosis with splenogastric hygropyrexia syndrome].

Splenogastric hygropyrexia syndrome, a common syndome in clinical practice, responds well to the therapy of traditional Chinese medicine (TCM). In view of the complexity of TCM syndrome formation, proceeding from different aspects such as Helicobacter pylori (Hp) infection, expression of inflammatory factors, mucosal protection and change in micro-ecosystem, the authors proposed that Hp infection in gastric mucosa, nuclear factor-kappa B activation, over-expression of heat shock protein 70, and "excessive evil, dyssplenism and confliction between healthy energy and evils" due to disequilibrium between Hp and lactobacillus acidophilus in gastric mucosa and coated tongue may be the important links in the formation of Hp related gastrosis with splenogastric hygropyrexia syndrome.

[Study on correlation between aquaporin 3, 4 gene expression in gastric mucosa and severity of Pi-Wei damp-heat syndrome in patients with chronic superficial gastritis].

OBJECTIVE: To explore the relationship between aquaporin 3,4 (AQP3, AQP4) gene expression in gastric mucosa and severity of Pi-Wei damp-heat syndrome (PWDHS) in patients with chronic superficial gastritis (CSG). METHODS: Gastric mucosa taken from the upper part of gastric corpus was collected under gastroscope and preserved in liquid nitrogen. The gene expression of AQP3 and AQP4 was determined quantitatively by fluorescent PCR. RESULTS: The gene expression of AQP3 and AQP4 in patients with PWDHS of moderate and severe degree was higher than that in those of mild degree and in healthy persons respectively (P <0.05 and P <0.01); and the gene expression of AQP3 in patients with PWDHS of severe degree was higher than that in those of moderate degree (P<0.05). CONCLUSION: The gene expression of AQP3 and AQP4 in gastric mucosa was correlative with the severity of PWDHS in patients with chronic superficial gastritis, the severer the syndrome, the higher the gene expression.

[Preliminary study on intestinal flora in diarrhea type irritable bowel syndrome with pi-wei dampness-heat syndrome].

OBJECTIVE: To observe the changes of intestinal flora in diarrhea type irritable bowel syndrome with Pi-wei dampness-heat syndrome (IBS-PDS). METHODS: The seven kinds of common intestinal bacteria in feces, including enteri bacillus, enterococci, saccharomycete, bifid bacteria, lactobacillus, bacteroides and peptococcus were studied in 21 patients suffered from IBS-PDS, and compared with those in 22 patients with IBS with deficiency of Pi syndrome (DPS) and 25 healthy subjects as control. RESULTS: As compared with the healthy subjects, the levels of enteri bacillus and enterococci were significantly increased (P<0.01), the levels of bifid bacteria, Lactobacillus and Peptococcus were significantly decreased (P < 0.01), and saccharomycete and Bacteroides were insignificantly different in patients with PDS. As compared with patients with DPS, the levels of enteri bacillus, enterococci, bifid bacteria, Lactobacillus, Peptococcus and Bacteroidaceae were significantly increased except the level of saccharomycete. CONCLUSION: There may be alteration of intestinal flora in patients with IBS-PDS.

Expression of bcl-2 oncogene in gastric precancerous lesions and its correlation with syndromes in traditional Chinese medicine.

AIM: To observe the protein and mRNA expression of bcl-2 oncogene in gastric precancerous lesions (GPL) and to analyze its correlation with syndromes in traditional Chinese medicine (TCM). METHODS: Sixty-seven patients with GPL confirmed by gastroscopy and pathology were studied, including 39 cases of moderate gastric mucosal dysplasia, 19 cases of severe gastric mucosa dysplasia, 9 cases of incomplete colon metaplasia. In syndrome differentiation of TCM, 17 cases belonged to the syndrome of qi and yin deficiency of the spleen and stomach complicated by qi stagnation, 21 cases belonged to the syndrome of qi and yin deficiency of the spleen and stomach complicated by stomach heat, 29 cases belonged to the syndrome of qi and yin deficiency of the spleen and stomach complicated by blood stasis. Protein and mRNA expression of bcl-2 oncogene were detected by labeled streptavidin biotin (LSAB) immunohistochemistry and in situ hybridization respectively. RESULTS: Abnormal expression of protein and mRNA on bcl-2 oncogene was found in GPL, which increased gradually with the course of lesions. In moderate and severe gastric mucosal dysplasia and incomplete colon metaplasia, there was no difference in the expression of bcl-2 oncogene (P>0.05). In different accompanying syndromes, the expression of protein and mRNA on bcl-2 oncogene increased gradually in the following order: deficiency of both qi and yin of the spleen and stomach accompanying qi stagnation-->stomach heat-->blood stasis. In GPL, compared with accompanying blood stasis, there was an obvious difference in the expression of bcl-2 oncogene between the syndrome of qi and yin deficiency of the spleen and stomach and accompanying stomach heat, so did accompanying qi stagnation (the level of protein: chi(2) = 8.45, P<0.05; the level of mRNA: chi(2) = 7.35, P<0.05). CONCLUSION: Apoptosis-associated bcl-2 oncogene is abnormally expressed in GPL, which correlates with different accompanying syndromes in TCM.

[Effect of Chinese herbs on expression of aquaporin 3,4 gene in gastric mucosa of patients with Pi-Wei Damp-Heat syndrome].

OBJECTIVE: To explore the relationship between Pi-Wei Damp-Heat Syndrome (PWDHS) with expression of aquaporin (AQP) 3,4 gene in gastric mucosa and the effects of Qingre Huashi Recipe (QHR) on the expression. METHODS: Sixty-eight patients with chronic superficial gastritis were differentiated into Pi-Wei Damp-Heat Syndrome group (PWDHS, n = 53, 19 cases with predominant Dampness, 14 cases with predominant Heat, 20 cases with Dampness equal to Heat) and Pi deficiency Syndrome group (PDS, n = 15). The PWDHS were treated with QHR. The expression of AQP 3,4 gene in the two groups were determined by fluorescence quantitative polymerase chain reaction (FQ-PCR). RESULTS: Expression of AQP 3 gene in PWDHS was higher than that in PDS and the healthy group, but the difference showed no statistical significance. Expression of AQP 4 gene in PWDHS was obvious higher than that in PDS and the healthy group (P <0.05 or P <0.01), but the difference of AQP 4 gene expression between PDS and the healthy group was insignificant. Comparison among various sub-types of PWDHS showed that the AQP 4 gene expression in the predominant dampness > dampness equal to heat> predominant heat. AQP 3,4 gene expression in PWDHS was significantly decreased after QHR treatment, especially in the cases with predominant dampness syndrome (P <0.01), approaching that in the healthy group and PDS. CONCLUSION: Abnormal expression of AQP 3,4 gene may be one of the possible mechanisms of PWDHS pathogenesis, Chinese herbs could influence AQP 3,4 gene expression to play a key role in treatment.

He Jie Tang in the treatment of chronic hepatitis B patients.

AIM: To explore the effect of He Jie Tang (decoction for medication) on serum levels of T lymphocyte subsets, NK cell activity and cytokines in chronic hepatitis B patients. METHODS: Eighty-five patients with chronic hepatitis B were divided randomly into two groups. Fifty patients in group I were treated with He Jie Tang (HJT) and 35 patients in group II were treated with combined medication. The levels of T-lymphocyte subsets (CD(3)(+), CD(4)(+), CD(8)(+)), NK cell activity, cytokines (TNF-alpha, IL-8, sIL-2R) were observed before and after the treatment. Another 20 normal persons served as group 3. RESULTS: The level of CD(4)(+) cells and NK cell activity were lower, whereas the level of CD(8)(+) cells in patients was higher than that in normal persons (t = 2.685, 3.172, and 2.754 respectively; P<0.01). The levels of TNF-alpha, IL-8, and sIL-2R in chronic hepatitis B patients were higher than those in normal persons (t = 3.526, 3.170, and 2.876 respectively; P<0.01). After 6 months of treatment, ALT, AST, and TB levels in the two groups were obviously decreased (t = 3.421, 3.106, and 2.857 respectively; P<0.01). The level of CD(4)(+) cells and NK cell activity were increased whereas the level of CD(8)(+) cells decreased (t = 2.179, 2.423, and 2.677 respectively; P<0.05) in group I. The levels of TNF-alpha, IL-8, and sIL-2R in group I were decreased significantly after the treatment (t = 2.611, 2.275, and 2.480 respectively; P<0.05) but had no significant difference in group II after the treatment (t = 1.906, 1.833, and 2.029 respectively; P>0.05). The total effective rate had no significant difference between the two groups (c2 = 2.882, P>0.05) but the markedly effective rate was significantly different between the two groups (c2 = 5.340, P<0.05). CONCLUSION: HJT is effective in treating chronic hepatitis B. HJT seems to exert its effect by improving the cellular immune function and decreasing inflammatory cytokines in chronic hepatitis B patients. The function of HJT in protecting liver function in the process of eliminating virus needs to be further studied.

Effect of Hejie decoction on T cell immune state of chronic hepatitis B patients.

AIM: To explore the effect of Hejie decoction (HJD) (mediation decoction) on T cellular immune state of chronic hepatitis B patients. METHODS: Sixty-five patients with chronic hepatitis B were randomly divided into 2 groups. Forty patients in the treatment group were treated by HJD, and 25 patients in the control group were treated by routine Western medicine. The TCRVbeta7 gene expression, T lymphocyte subsets (CD3+, CD4+, CD8+, CD4+/CD8+) levels were observed before and after treatment. RESULTS: The level of CD4+ cells was lower whereas the level of CD8+ cells was higher in patients than in the normal group. There was no significant difference between the levels of CD3+ cells in patients and normal persons. After 6 months of treatment, ALT, AST, TB levels of the 2 groups were obviously decreased, and the level of CD4+ cells was increased whereas the level of CD8+ cells was decreased in the treatment group. However, the level of CD4+ cells and CD8+ cells had no significant difference in the control group. TCRVbeta7 expressions were detected in 6 patients of the treatment group, whose HBV-DNA and HBeAg turned negative and ALT became normal. HBeAg in another 3 patients turned negative while HBV-DNA did not, and TCRVbeta7 expressions were not detectable. TCRVbeta7 expression could not be detected in the control group, HBV-DNA of the control group did not turn negative. HBeAg in 1 patient turned negative while HBV-DNA did not, and TCRVbeta7 expressions were not detectable. The total effective rate was not significantly different between the 2 groups and the markedly effective rate was significantly different (P<0.01). CONCLUSION: HJD is effective for treating chronic hepatitis B, and its effect seems to relate with the improvement of the TCRVbeta7 expression of chronic hepatitis B patients, thus activating T cells and eliminating HBV. T cellular immune function plays an important role in HBV infection and virus elimination.