WANG Xinlu's Experience in Treating Metabolic Syndrome based on “Blood Turbidity” / 中医杂志

This paper summarized professor WANG Xinlu's experience in treating metabolic syndrome （MS） based on the “blood turbidity” theory. It is believed that blood turbidity is the key pathological factor for the onset of MS. Blood turbidity accumulates internally， and the zang-fu （脏腑） organs become useless， leading to MS. With long-term blood turbidity， phlegm and stasis are cemented， and the condition is worsened， having changed syndromesfrequently. In clinical practice， the basic treatment method is to clear and dissolve blood turbidity， taking self-prepared Modified Huazhuo Xingxue Decoction （加味化浊行血汤） as the basic formula， and flexibly modifying it according to different stages and manifestations of the disease. Simultaneously， supplementary modern medicines are used to treat abnormal indicators of western medicine.

Papillon-Lefèvre syndrome: a case report and mutation analysis of the cathepsin C gene / 中华皮肤科杂志

Objective To analyze mutations in the cathepsin C (CTSC) gene in a patient with Papillon-Lefèvre syndrome (PLS).Methods Clinical data were collected from a patient with PLS.Two milliliters of venous blood samples were obtained from the patient,his parents and 100 unrelated healthy controls separately.DNA was extracted from these blood samples,and PCR was performed to amplify all the 7 exons of the CTSC gene followed by direct DNA sequencing.Results Two heterozygous mutations were observed in the CTSC gene of the patient.One was a novel mutation c.824C ＞ T at position 824 in the exon 6,which resulted in a substitution of ACC (threonine) by ATC (isoleucine) at codon 275 (p.T275I).The other one was the mutation c.1040A ＞ G at position 1040 in the exon 7,causing the substitution of TAT (tyrosine) by TGT (cysteine) at codon 347 (p.Y347C).His father and mother carried the heterozygous mutation c.824C ＞ T and c.1040A ＞ G respectively.Neither of the two mutations was observed in the 100 healthy controls.Conclusions CTSC mutations are responsible for the clinical phenotype of PLS.Identification of the c.824C ＞ T mutation extends the spectrum of mutations in the CTSC gene and provides a basis for genetic diagnosis of PLS.

A case of keratitis-ichthyosis-deafness syndrome complicated by cutaneous squamous cell carcinoma：mutation analysis of the GJB2 gene and literature review / 中华皮肤科杂志

Objective To analyze mutations in the GJB2 gene in a Chinese patient with keratitis-ichthyosis-deafness (KID)syndrome complicated by cutaneous squamous cell carcinoma. Methods Clinical data were collected from a patient with KID syndrome complicated by cutaneous squamous cell carcinoma. Peripheral blood samples were obtained from the patient and her parents, and DNA was extracted from these blood samples. PCR was performed to amplify the exon 2 of the GJB2 gene followed by direct DNA sequencing. Results A mutation (c.148G > A)was identified at position 148 in exon 2 of the GJB2 gene, which caused a codon change from GAC to AAC and resulted in the substitution of aspartate by asparagine at position 50 in the connexin26 (Cx26)protein (p.Asp50Asn). Inaddition,anothermutation(c. 79G > A), which led to the substitution of valine by isoleucine at codon 27 in Cx26 (p.Val27Ile), was found at position 79 in exon 2 of the GJB2 gene. Neither of the two mutations was detected in the patient′s parents. Literature review revealed that 13 cases of KID syndrome complicated by cutaneous squamous cell carcinoma had been reported in abroad, and the mutation c.148G > A was detected in the GJB2 gene in all the 7 cases finally diagnosed by gene sequencing. Conclusion GJB2 gene mutations may be responsible for the clinical phenotype of KID syndrome in this Chinese patient, and the mutation c.148G > A may be related to the development of cutaneous squamous cell carcinoma.

Benign familial chronic pemphigus in a family: a clinical survey and mutation analysis of ATP2C1 gene / 中华皮肤科杂志

Objective To report a Chinese pedigree with benign familial chronic pemphigus (BFCP),and to screen mutations of ATP2C 1 gene in this family.Methods A 39-year-old male patient with BFCP andhis family members underwent a clinical investigation.Blood samples were collected from all the members in this family and from 50 unrelated healthy controls.Genomic DNA was extracted from the blood samples,and PCR was performed to amplify all the 28 exons and flanking sequences of the ATP2C1 gene followed by DNA direct sequencing.The resulted DNA sequences were compared with the reported sequences of APT2C1 gene in Genbank (Number:NM_014382.2 and NC_000003.9).Results There were 24 family members in the four-generation pedigree,with 8 members affected by BFCP.A single-nucleotide substitution,c(1696C→T),in exon 17 of the ATP2C1 gene was identified in all of the members with BFCP,but not in unaffected third-or second-generation members or unrelated healthy controls.This substitution was also found in 1 out of 4 family members of fourth-generation.Conclusions The nonsense mutation c(1696C→T) in the ATP2C1 gene,is likely to be responsible for BFCP in this Chinese four-generation pedigree.The underage family member of fourth-generation who carried the mutation c(1696C→T) but had no clinical symptoms of BFCP,should be closely followed.

Heat shock protein 110 improves the immunological effect of an altered peptide ligand of human papilloma virus type 16 E711-20 peptide / 中华皮肤科杂志

ObjectiveTo investigate the adjuvant effect of heatshock protein 110(HSP110) on the immune responses induced by an altered peptide ligand of human papilloma virus type 16 E711-20 peptide (HPV16E711-20).Methods The complex of HSP110 and an altered peptide ligand of HPV16E711-20 was constructed in vitro.Fifteen 6-week-old C57BL/6 female mice were randonly and equally divided into 3 groups,including complex group,ligand group,and phosphate buffered solution (PBS) group,to receive intraperitoneal immunization with the complex (100 μg),peptide (10 μg),and PBS (100 μl) respectively.Immunization was carried out with an interval of 2 weeks for 2 times.Two weeks after the last immunization,the mice were sacrificed followed by the isolation of splenocytes.MTT assay was performed to evaluate the proliferation activity of splenocytes,intracellular staining for interferon(INF)-γ to detect cytotoxic T lymphocytes (CTLs),standard chromium-51 (51Cr) release assay to estimate the lethal effect of specific CTLs on target cells.Statistical analysis was performed by using t test with SPSS 10.0 software.Differences were considered as statistically significant when the P value was less than 0.05.ResultsA significant increase was observed in the proliferation index ( 1.87 ± 0.122 vs.0.32 ± 0.071,t =4.01,P ＜ 0.01 ) of,and percentage of CD8+IFN-γ+ T lymphocytes(3.9％ vs.0.4％,t =3.88,P ＜ 0.01 ) among splenocytes from the complex group compared with the ligand group.At the effector-to-target ratio of 100 ∶ 1,50 ∶ 1,25∶ 1 and 12.5 ∶ 1,the death rate of target cells was 54.7％,72.2％,61.5％ and 39.8％ respectively after incubation with CTLs from the compleximmunized mice,higher than that from the ligand-immunized mice (35.2％,49.3％,28.1％,17.4％,respectively).ConclusionHSP110 could enhance the immunological effect of the altered peptide ligand of HPV16E711-20,and can serve as an immunological adjuvant.

Determination of human papillomavirus types in giant condyloma acuminatum / 中华皮肤科杂志

ObjectiveTo determine the genotypes of human papillomavirus(HPV) in patients with giant condyloma acuminatum.MethodsSixty-seven outpatients with giant condyloma acuminatum collected from January 2007 to January 2010 were included in this study.Lesional specimens were obtained from these patients.The genotypes of HPV were determined by flow-through hybridization and gene chip assay.ResultsOf the 67 cases of giant condyloma acuminatum,63 (94.02％) were positive for HPV DNA.Among the HPV DNA-positive specimens,84 (60.87％) harbored low risk types of HPV,54 (39.13％) high risk types of HPV.Type 6 and 11were the predominant low risk HPV types,while type 16 and 18 were the major high risk HPV types.

Immunogenicity of the immunodominant cytotoxic T lymphocyte epitope E749-57 in HPV16 oncoprotein E7 chaperoned by HSP110 / 中华皮肤科杂志

Objective To investigate the immunogenicity of immunodominant cytotoxic T lymphocyte epitope E749-57 of human papilloma virus (HPV) 16 oncoprotein E7 chaperoned by heat shock protein (HSP)110. Methods Mouse HSP110 gene was cloned into prokaryotic expression vector pQE-80L for the expression of HSP110 protein, which was purified using Ni-NTA column. SDS-PAGE and Western-blot were conducted to confirm the purified mHSP110 protein, which was subsequently incubated with E749-57 peptide under heat shock condition, and high-performance liquid chromatography (HPLC) was used to evaluate the binding efficiency of the recombinant protein and E749-57 peptide. Twenty mice were divided into 4 groups to be immunized with mHSP110 protein, E749-57 peptide, mHSP110-E749-57 complex and phosphate buffered saline (PBS),respectively. Two weeks after the last immunization, spleen cells were collected from the immunized mice and divided into 2 parts: one were stimulated by E749-57 peptide followed by the detection of CD8+ INF-γ+ T cells with flow cytometry; the other one were subjected to MTT analysis for the estimation of cell proliferation. The mHSP110-E749-57 complex was also used to immunize TC-1 tumor bearing mice to observe its anti-tumor effect.Results The full-length 2577 bp-sized mHSP110 gene was amplified from mouse liver cDNA and cloned into pQE-80L vector. Direct sequencing confirmed the correctness of the cloning. SDS-PAGE and Western-blot demonstrated the successful purification of mHSP110. HPLC assay showed that the purified mHSP110 protein could bind with E749-57 to form a relatively stable protein complex. The percentage of IFN-γ+ CD8+ T cells in and proliferation index of spleen cells from the complex-immunized mice were statistically higher than those from the other 3 groups of mice. Moreover, the complex could obviously inhibit the growth of TC-1 tumor in mice. Conclusion The mHSP110-E749-57 complex could enhance the generation of specific cytotoxic T lymphocytes and exert anti-tumor effects in mice.

Tiaohe Ganpi Hexin Decoction in treatment of irritable bowel syndrome with diarrhea: a randomized controlled trial / 中西医结合学报

Background: The quality of life has been greatly influenced and the cost of medical expenses is very high in patients with irritable bowel syndrome (IBS). The etiology and pathogenesis of IBS are still unclear, and the prevention and treatment of this disease still lack of effective methods. Objective: To explore and analyze the effects of Tiaohe Ganpi Hexin Decoction (TGHD), a compound traditional Chinese herbal medicine for regulating the liver and spleen, on IBS patients with diarrhea. Design, setting, participants and interventions: All 40 IBS patients came from the First Hospital of Jinan University, Guangzhou Red Cross Hospital, and the First Affiliated Hospital of Guangzhou University of Chinese Medicine, and were randomly divided into two groups. Patients in the treatment group (n=20) were given TGHD, while those in the control group (n=20) were prescribed oral pinaverium with a four-week treatment period. Main outcome measures: Traditional Chinese medicine (TCM) syndrome score, total obviously effective rate, disappearance rate of symptoms, and clinical symptom score in the two groups were evaluated before and after four-week treatment. Results: After the treatment, TCM syndrome scores in both groups were decreased (P<0.01), and the TCM syndrome score in the treatment group was significantly lower than that in the control group (P<0.01). There was a significant difference in the total obviously effective rate between the two groups (P<0.01), and the total obviously effective rates in the treatment and control groups were 85%(17/20) and 45%(9/20) respectively. The disappearance rates of abdominal pain, abdominal distention, poor stool output, stool frequency, stool character and mucous stool in the treatment group were higher than those in the control group (P<0.05). The symptom scores of abdominal pain, abdominal distention, poor stool output, stool frequency, stool character and mucous stool in the treatment group were lower than those in the control group (P<0.05). Conclusion: TGHD can significantly improve the clinical symptoms in IBS patients with diarrhea.

Qiongyugao for the bone marrow inhibition of mice with pulmonary adenocarcinoma / 中国组织工程研究

BACKGROUND: Bone marrow inhibition is a predominant side effect caused by the drugs for anti-tumor.OBJECTIVE: To observe the intervention effect of traditional Chinese medicine qiongyugao on bone marrow inhibition induced by chemotherapy in experimental mice with pulmonary adenocarcinoma.DESIGN: Randomized controlled trial.SETTING: Medical College of Jinan University.MATERIALS: The experiment was completed in the Central Laboratory of Medical College of Jinan University from March to October in 2001. Fortyeight healthy, aged 6 to 8 weeks C57/BL mice were involved.METHODS: The mouse model with pulmonary adenocareinoma was made with the method of inoculability of cancer cell, and then the models were group: On the following day of inoculation, 0.2 mL normal saline was used by gavage, and normal saline was injected intraperitoneally at the dosage of amine 20 mg was diluted into 0.2 g/L with normal saline, and performed intraperitoneal injection at the dosage of 5 μL/g, once per day, and 0.2 mL platinum diamine was used as in chemotherapy group and 0.2 mL qiongyugao was administered by gavage at the same time once per day. Peripheral erythrocyte, leukocyte and blood platelet, and karyota in bone marrow counting were detected on the 21 days after administration.MAIN OUTCOME MEASURES: Peripheral erythrocyte, leukocyte and blood platelet, and karyota in bone marrow counting of the mice in each group.RESULTS: Eleven mice failing to have tumor in tumor assessment were excluded from the sample, including 3 in the control group, 4 in the chemotherapy and 4 in the combination group. During the experiment,there were 2 mice dying in each group of the control group and chemotherapy group. Totally 11 mice in the control group, 10 in the chemotherapy group and 11 in the combination group entered the result analysis. Erythrocyte, leukocyte and blood platelets in the combination group and control group were all significantly higher than those in the chemotherapy group [erythrocyte: (8.54 ±0.81 ), (8.65 ±0.77), (4.56 ±1.00) ×1012 L -1;leukocyte:(9.04 ±0.60), (9.14 ±0.71), (3.31 ±0.96) ×109 L -1;blood platelets:(949.09±111.31 ), (955.54±87.13), (399.30±131.36)× 109 L-1 ,P ＜ 0.01]. The number of karyota in bone marrow in the chemotherapy group was significantly lower than those in the combination group and control group [(5.30±1.12),(10.51±1.15),(14.36±1.02)]×106,P ＜ 0.01); while the number of karyota in the control group was higher than that in the combination group (P ＜ 0.05).CONCLUSION: Qiongyugao can enhance the counting of erythrocyte,leucocyte, blood platelets of peripheral blood and karyota in bone marrow after chemotherapy in mice with pulmonary carcinoma, improve the inhibition of bone marrow induced by chemotherapy, but can not reverse it completely.

Study on mechanism of action of promoting blood circulation and dispersing phlegm method in improving insulin resistance of type 2 diabetes rats / 中华中医药杂志

0.05).Compared with control group,the GK activity of liver cell,the expression of PEPCK and the expression of GLUT4 in model group decreased signifi cantly(P

Effect of Guizhi Decoction on cardiac muscular collagen reconstruction in spontaneous diabetes rats / 中华中医药杂志

Objective:This study was to study effect of keeping ying and weiqi in balance on cardiac muscular collagen Ⅰ,collagen Ⅲ,angiotensinⅡ in spontaneous diabetes rats,and provide new thoughts for prevention and treatment for diabetic cardiomyopathy.Methods:40 spontaneous diabetes rats were divided into four groups at random:spontaneous diabetes control group,diformin group,Guizhi Decoction group and Huanglian Jiedu Decoction group.Except control group,each group was given intragastric administration for 12weeks.Blood glucose,collagenⅠ,collagen Ⅲ,collagenⅠ/collagen Ⅲ,angiotensin Ⅱ were determined.Euzymelinked immunosorbent assay and radio-immuniy method were used.Results:Blood glucose in Huanglian Jiedu Decoction group and diformin group decreased;Compared respectively with diformin group and control group,collagenⅠdecreased obviously and collagen Ⅲ increased signifi cantly in Guizhi Decoction group,and the collagenⅠ/collagen Ⅲ decreased.Compared with diformin group and control group,cardiac muscular homogenate AT-Ⅱ contents decreased in both of Guizhi Decoction group and Huanglian Jiedu Decoction group.Conclusion:Guizhi Decoction can decrease the content of collagen Ⅰ,AT-Ⅱ and collagenⅠ/collagen Ⅲin cardiac muscular,and increase content of collagen Ⅲ,then prevent and treat cardiac muscular collagen reconstruction and myocardial fibrosis.Huanglian Jiedu Decoction can decrease the content of AT-Ⅱ,and decrease collagenⅠ/ collagen Ⅲ,collagenⅠ slightly,and can improve cardiac muscular collagen reconstruction.Keeping ying and weiqi in balance is a new thought for prevention and treatment for diabetic cardiomyopathy.

Clinical Significance of Nucleosomes Released from Peripheral Blood Mononuclear Cells of Patients with Systemic Lupus Erythematosus / 中华皮肤科杂志

Objective To assess the levels of nucleosomes released from peripheral blood mononu-clear cells(PBMC)of patients with systemic lupus erythematosus(SLE),and their relationship with auto-antibodies as well as disease activity.Methods Levels of both nucleosomes released from PBMC and vari-ous auto-antibodies were detected by ELISA in sera from SLE patients.The disease severity was evaluated using SLEDAI(systemic lupus erythematosus disease activity index)system.Results Levels of nucleosomes released from PBMC were significantly higher in patients with active SLE than those of patients with inactive disease and normal controls(39.39?25.70,13.44?8.82,and11.73?7.87IU/mL,respectively).There was a significant positive correlation between nucleosome levels and SLEDAI scores,serum ds-DNA auto-an-tibody levels,and low C3levels.Conclusion Nucleosomes released from apoptotic PBMC of patients with SLE is closely correlated to disease activity,which implies that nucleosomes may play an important role in the pathogenesis of SLE.

Isolation, purification and identification of nucleosomes from chicken erythrocytes / 第三军医大学学报

Objective To establish a simple and practical method for isolating and purifying nucleosomes. Methods Nuclei were isolated from chicken erythrocytes, and then digested with staphylococcal nuclease. After centrifugation, the supernatant of digestion was separated and centrifugated on sucrose gradients. Results Nucleosomes with good stability were isolated properly by gradient centrifugation. Conclusion This method for the isolation and purification of nucleosomes is simple and effective, which might contribute to the further researches of the roles of nucleosomes in the pathogenesis of systemic lupus erythematosus.

Antigen-specific Cellular Immune Response Induced by Dendritic Cells Activated by Human Papilloma Virus 16 E7 Peptide / 中华皮肤科杂志

Objective To investigate specific cellular immune response induced by dendritic cells(DCs) activated by human papilloma virus (HPV) 16 E7 peptide. Methods DCs separated from peripheral blood mononuclear cells were induced with granulocyte/macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4), the DCs were then activated by HPV 16 E7 peptide. After that the surface antigens on the DCs were detected by flow cytometer. Homogenic mixed lymphocyte reaction (HMLR) with DCs was performed. LDH release assay was also used to detect the activity of cytotoxic T lymphocytes(CTL)to Caski tumor cells, which was induced by DCs. Results The surface antigens on the activated DCs were highly expressed, such as CD1a(58.4 ? 6.7)%, CD80(70.6 ? 3.4)% and HLA-DR(74.8 ? 4.2)%. HMLR showed that the activated DCs stimulated T cell proliferation significantly. Antigen-specific CTL induced by the activated DCs specifically killed Caski tumer cells, but had no effect on SiHa and AN3CA cells. Conclusions DCs loaded with HPV 16 E7 peptide can induce highly effective and specific cellular immune response.

Nucleosome Induced Systemic Lupus Erythematosus-like Disease in BALB/c Mice / 中华皮肤科杂志

Objective To investigate the role of nucleosome in the pathogenesis of systemic lupus erythematosus (SLE). Methods BALB/c mice were immunized with nucleosome, and then serum dsDNA and ANA autoantibodies were detected by ELISA. Kidney specimens were observed by immunofluorescence and histological examination. Results High titers of IgG dsDNA and ANA autoantibodies in sera of BALB/c mice were observed at the 14th day after immunization with nucleosome. Nephritis and immune complex deposition in renal glomeruli were observed at the 35th day. Conclusion Nucleosome could induce SLE-like disease in BALB/c mice, and may play a critical role in the pathogenesis of SLE.

Screening and Identification of Predicted Epitopes of HLA-A2-restricted Cytotoxic T Lymphocytes Derived from the HPV16 E7 Antigen / 中华皮肤科杂志

Objective To screen and identify the predicted epitopes of synthesized predicted HLA-A2-restricted cytotoxic T lymphocytes (CTLs) derived from HPV16 E7 antigen. Methods The predicted epitopes of HLA-A2-restricted CTLs derived from HPV16 E7 antigen were synthesized and purified with Standard Fmoc assays, and the standard 51Cr release assay was used to determine their activities to induce specific CTL. Results Two epitopes of HLA-A2-restricted CTLs, namely E711-19 (YMLDLQPET) and E749-57 (RAHYNIVTF) derived from HPV16 E7 antigen were identified. Conclusion E711-19 (YMLDLQPET) and E749-57 (RAHYNIVTF) have antigenicity, and may be the candidates for development of peptide vaccine in the treatment of HPV infections.

Molecular Modification and Identification of HPV16E749-57 / 中华皮肤科杂志

Objective To introduce the amino acid substitution for HP V16E749-57(HLA-A2-restricted CTL epitope) and to identify the novel epitopes.Me thods Quantitative method was used to evaluate the affinity of the substituted peptides.To determine the peptide candidates to be synthesized and identified,the molecular models of the HLA-A2-peptide complex and CTL epitope candidates b ound to the HLA-A2 molecule were established by computer molecular modeling.Pep tides were synthesized and purified with standard Fmoc assay,lactate dehydrogen ase (LDH) release assay was used to determine their abilities of inducing the ge neration of specific CTLs.Results Modified peptides met the requirements of H LA-A2-restricted CTL epitopes.Peptide RLHYNIVTF had the abilitiy of inducing th e generation of specific CTLs.Conclusions Compared with HPV17E749-57 the mod ified peptide RLHYNIVTF has a higher antigenicity and affinity to HLA-A2.So,pe ptide RLHYNIVTF may be used as one of the HLA-A2-restricted candidate epitopes,instead of HPV17E749-57,for peptide vaccine in the treatment of HPV infection.

Effects of Keningfang decoction on heat shock protein 70 expression in lung of mice with influenza virus pneumonia / 中国病理生理杂志

AIM: To study the preventive effects of Keningfang decoction on the experimental influenza virus pneumonia in mice and its mechanism. METHODS: Fifty NIH mice were divided into five groups randomly (ten mice in each group), normal control group, model group, virazole treatment group, Keningfang I treatment group, Keningfang II treatment group. The FM 1 virus strain that kept in frozen condition were revived and cultured in chick embryo. The mice in every group that were lightly anesthetized by aether, and infected by dropping FM 1 15 LD 50 into the nose, except for the normal control group, by equal volume distilled water. Mice were treated with drugs or distilled water two days before the model was made (0 3 mL, 2 times a day). The mice were treated with drug for six days, then was killed, the lungs were collected, and kept in -70 ℃. HSP70 was measured in the lung tissue by Western blot. Pathologic changes of the mice lungs were observed under microscope. RESULTS: Compared with the normal control group, HSP70 in the other groups were increased significantly (P