AIM: To explore the effects of tumor necrosis factor-? (TNF-?) on inducible nitric oxide synthase (iNOS)-nitric oxide (NO) system activity in arginine vasopressin (AVP)-induced rat cardiac fibroblasts (CFs). METHODS: CFs were isolated by trypsin digestion method. Nitrate reductase method, spectrophotometry and reverse transcription-polymerase chain reaction (RT-PCR) were used to detect NO contents, NOS activity and iNOS mRNA expression, respectively. RESULTS: AVP significantly increased iNOS mRNA expression, NOS activity and NO contents in CFs. Combined with AVP, TNF-? enhanced the effects of AVP on iNOS-NO system activity in a concentration-dependent manner. However, if the concentration of TNF-? was too high, the iNOS-NO system activity did not increase accordingly, but slightly decreased instead. CONCLUSION: TNF-? stimulates iNOS-NO system activity in coordination with AVP in CFs. The enhancement of NO contents inhibits ventricular remodeling induced by AVP and TNF-?. [

Objective To explore the effect of arginine vasopressin(AVP) on proliferation and nitric oxide(NO) synthesis in rat cardiac fibroblasts(CFs). Methods CFs were isolated by trypsin digestion method. The effect of AVP on proliferation and NO synthesis were measured by MTT technique and nitric acid reductase method respectively. Results The MTT OD values of CFs increased in a concentration-dependent manner with the rising of the concentration of AVP and in a time-dependent manner with the prolongation of the culture time. For example, the MTT OD values of 10 -6 mol/L AVP group were higher than those of the control, and the MTT OD values of CFs cultured for 36h were significantly higher than those cultured for 6h when intervened with 10 -7 mol/L AVP. The content of NO of CFs displayed the same change as the MTT OD values. The contents of NO in 10 -7 mol/L and 10 -6 mol/L AVP groups were statistically higher than those of the contral group, 10 -9 mol/L AVP group and 10 -8 mol/L AVP group. The NO contents of CFs cultured for 24h and 36h were significantly higher than those cultured for 6h and 12h. Conclusion AVP can stimulate the proliferation of CFs and the synthesis of NO in neonatal rat. The enhancement of NO contents would inhibit CFs proliferation and myocardial fibrosis induced by AVP.

Objective:To compare the efficacy and adverse reactions between intralesional betamethasone injection combined with botulinum toxin type A and topical hyaluronic acid and simple intralesional betamethasone injection and topical hyaluronic acid in treatment of keloid.Methods:A total of 58 patients with keloid were divided into two groups: intralesional betamethasone injection combined with botulinum toxin type A and topical hyaluronic acid group (combined group, n=28) and intralesional betamethasone injection combined with topical hyaluronic acid group (control group, n=30).In both two groups, the intralesional betamethasone injection was given once every 4 weeks for 3 times totally, and topical hyaluronic acid was applied every day.In combined group, an injection of botulinum toxin type A was given around the lesion after the first intralesional injection of betamethasone.The parameters for therapeutic efficacy and adverse reactions of the patients in two groups were recorded, and the clinical pictures were taken before and after each treatment, and they were analyzed and compared.Results:After 3 treatments, compared with control group, the lesion appearance in combined group was improved.In control group, the VAS value was reduced at 1 month, but was increased gradually at 2 and 3 months. In combined group, the VAS value was reduced gradually in 3 months. There was a significant difference in VAS value at 2 and 3 months between two groups (P<0.05).In combined group, the thickness of keloids were reduced gradually in 3 months, while the reduction was not obvious in control group (P<0.05).The recurrence of pain, itching and lesion height were observed 2 weeks after each treatment in control group, but there was no recurrence in combined group.The incidence rate of adverse reactions was 26.7% in control group, which had no significant difference compared with combined group(25.0%) (P>0.05).Conclusion:Compared with control group, the combined group shows better efficiency in the treatment of keloid and there is no increase of adverse reactions.The therapy is worthy of clinical application.

Objective To investigate the value of the expressions of estrogen receptor(ER)and progesterone receptor(PR)in the cast-off cells from the fluid of latex ductal lavage in the diagnosis and treatment of breast diseases.Methods A total of 58 female out-patients were examined by fiberoptic ductoscopy(FDS).All patients were divided into 6 groups by the complaints,physical examination,ultrasound and FDS:normal group(10 persons without symptoms and abnormalities in any examination),galactostasis group(n=19),latex fill up group(n=11),intraductal papilloma group(n=2),mammary ductal ectasia group(n=7)and contralateral examination after mastectomy for breast cancer group(n=9).A total of 168 ducts were successfully examined by FDS,and the expressions of ER and PR in the cast-off cells were detected among all of the samples.Results The positive rates of ER and PR expression in the cast-off cells from the fluid of latex ductal lavage in benign breast disease groups were much higher than those in normal group(P

The points, Neiguan (PC 6)-Gongsun (SP 4), Houxi (SI 3)-Shenmai (BL 62), Lieque (LU 7)-Zhaohai (KI 6), or Waiguan (TE 5)-Zulinqi (GB 41), were selected according to the eight magic turtle techniques and some acupoints were added on the basis of syndrome differentiation. Thirty obesity patients were treated. Marked effectiveness occurred in 6 cases, effectiveness in 21 cases and ineffectiveness in 3 cases, with a total effective rate of 90.0%.

Aim To investigate the effects of cyclosporin A (CsA) on growth and collagen synthesis of cardiac fibroblasts (CFs) induced by arginine vasopressin (AVP). Methods CFs of neonatal Sprague-Dawley rats were isolated by trypsinization and cultured; growth-arrested CFs were stimulated with 1×10-7 mol·L-1 AVP in the presence or absence of CsA (0.05, 0.5 and 5 μmol·L-1). MTT and flow cytometry techniques were adopted to measure cell number and analyze cell cycle respectively. Collagen synthesis was determined by measurement of hydroxyproline content in culture supernatant with colorimetry. Calcineurin activity was estimated by chemiluminescence. Trypan blue staining to test the viability of CFs. Results 0.05, 0.5 and 5 μmol·L-1 CsA inhibited the increase of CFs number induced by 1×10-7 mol·L-1 AVP in a dose-dependent manner, with the inhibitory rates by 12%, 24% and 29%, respectively (P＜0.05). Furthermore, cell cycle analysis showed 0.5 μmol·L-1 CsA decreased the S stage percentage and proliferation index of CFs stimulated by AVP (P＜0.05). In culture medium, the hydroxyproline content induced by AVP decreased by 0.5 and 5 μmol·L-1 CsA (P＜0.05), with the inhibitory rates of 29% and 33%, respectively. CsA completely inhibited the increment of calcineurin activity induced by AVP (P＜0.01), but CsA itself had no effect on the baseline of calcineurin activity and CFs viability. Conclusion CsA inhibits proliferation and collagen synthesis of CFs by virtue of blocking calcineurin signaling pathway and might provide a novel target for prevention and treatment to cardiac fibrosis.

Objective: To evaluate the clinical therapeutic efficacy and safety of red-blue light irradiation combined with Xihuang Capsules in the treatment of moderate to severe acne vulgaris.Methods:Eighty patients with moderate to severe acne vulgaris were randomly divided into Xihuang Capsules group (control group)and red-blue light irradiation combined with Xihuang Capsules group (observation group).The patients in observation group were treated with red-blue light irradiation (20 min biw,4 weeks)combined with Xihuang Capsules (1.5 g bid,4 weeks),while the patients in control group took only Xihuang Capsules orally (1.5 g bid,4 weeks),The changes of skin lesions and adverse reactions were observed and the curative efficacies were compared at 2 or 4 weeks after treatment.Results:After 2-week-treatment,the total effective rate in observation group was 67.5% while the total effective rate in control group was 52.5%,there was no statistically significant difference between two groups (P >0.05).The regression number of inflammatory and non-inflammatory skin lesions in observation group was significantly higher than that in control group (P <0.05).After 4-week-treatment,the total effective rate in observation group was significantly higher than that in control group (85.0% vs 65.0% ),and the difference was statistically significant (P < 0.05);the regression number of inflammatory and non-inflammatory skin lesions in observation group was significantly higher than that in control group (P <0.05).During the treatment,there were no severe adverse reactions in two groups.Mild headache and nausea occurred in observation group after red-blue light irradiation,but the above symptoms disappeared after reducing the irradiation dose.Conclusion:Red-blue light irradiation combined with Xihuang Capsules in the treatment of moderate to severe acne is proved to be a significantly effective and safe method,and it is worth to be promoted in clinic.

Objectives The cellular repressor of E1A-activated genes (CREG), a novel gene, was recently found to play a role in inhibiting cell growth and promoting cell differentiation. The purpose of this study was to obtain antibody against CREG protein and to study the expression of CREG protein in human internal thoracic artery cells (HITASY) which express different patterns of differentiation markers after serum withdrawal. Methods The open reading frame of CREG gene sequence was amplified by PCR and cloned into the pGEX-4T-1 vector. Glutathione-S-transferase (GST)-CREG fusion protein was expressed in E. Coli BL21 and purified from inclusion bodies by Sephacryl S-200 chromatography. Rabbits were immunized with the purified GST-CREG protein. Western blot examined with immunohistochemistry staining and the protein expression level was analyzed by Western blot in HITASY cells after serum removal. Results It was confirmed by using endonuclease digesting and DNA sequencing that the PCR product of CREG was correctly inserted into the vector. The GST-CREG protein was purified with gel filtration chromatography. Polyclonal antibody against GST-CREG was obtained from rabbits. CREG protein immunohistochemistry staining displayed a perinuclear distribution in the cytoplasm of HITASY cells. Results from Western blot suggested that comparing with the untreated cells upregulation of CREG polyclonal antibody against CREG was comfirmed. Using this antibody, the changes of CREG protein expression was observed in the process of phenotypic modulation of HITASY cells. These results provide basic understanding on the relationship of CREG gene with the cell phenotypic conversion.

OBJECTIVE To prove the diagnosis value for Legionnaires pneumophial pneumonia using polymerase chain reaction. METHODS L. pneumophial-DNA (LPN-DNA) from 47 spuum and 6 bronchoalveolar lavage fluid samples collected from 53 patients with atypical pneumonia was detected by PCR. RESULTS The positive rate of LPN-DNA in 53 patients with atypical pneumonia was 9.4%, while the positive rate of sputum and bronchoalveolar lavage fluid samples was 6.4%and 33.3%, respectively. CONCLUSIONS LPN-DNA detected by PCR for early diagnosis of atypical pneumonia has favorable clinical application.

Objective To detect whether mice bone marrow mesenchymal stem cells(BMSCs)can contribute to the regeneration of hepatocytes in bone marrow chimeric mice.Methods Female recipient mice(C57BL/6J)underwent whole body gamma-ray irradiation with a dose of 10 Gy to ablate their bone marrow,followed by immediate tail vein injection of BMSCs isolated from male GFP transgenic mice.Animals were killed at different phase points:1 week,1 month,and 3 months.Using fluorescence microscope we directly observed GFP-positive cells in the liver frozen sections,and we also prepared the parafilm sections to detect the GFP-positive cells and the coexpression of GFP and Alb,CK18 by immunohistochemistry and immunofluorescence respectively.Results We found numerous GFP-positive cells in recipient mice liver at 1 week after BMSCs transplantation,some at 1 month and seldom at 3 months.There were some cells coexpressing GFP and Alb,CK18 at all the phase points.Conclusion Allotype BMSCs can differentiate into Alb and CK18 positive hepatocytes in bone marrow chimeric mice,which will become an ideal cell resource for liver tissue project.