Yishen Juanbi Method for Treating Rheumatism began from Mr. ZHANG Ci-gong to the formation and development of Mr. ZHU Liang-chun, and by the evidence-based source, innovation and perfection of the third and fourth generations of successors, has formed the unity of the principle, rule, method, medicine, and the unique diagnosis and treatment of rheumatism of integration of the internal treatment and external treatment. All of these provide original insights for rheumatology of TCM theory and the application of insect medicine, and provide valuable treatment technology for China's young rheumatology disciplines.

Objective:To construct the Helicobacter pylori infected C57BL/6 mice model to observe the activation of NOD1/NF-κB signaling pathways in the gastric tissues,and study its roles in inflammatory response during Hp infection.Methods:6-8 week-old C57BL/6 mice were randomly divided into two groups,the Hp infection group and the control group,and mice were given by gavage every 48 h for five times with Hp or PBS,respectively.All the animals were sacrificed at different time point and the gastric tissue were stained with hematoxylin-eosin( HE);The mRNA expression of NOD1 and RIP2 in gastric tissues were examined by RT-PCR;Levels of IFN-βand IP-10 in mice serum were assessed by ELISA;Nuclear translocation of p65 in gastric tissue was detected by Western blot.Results:Hp infection elicits an inflammatory cell response,glands in gastric tissue were reduced or atrophic,as compared with that in the control group.The levels of IP-10 and IFN-βincreased in the model group, and peaked at 16 weeks after Hp infection.Hp infection increased the mRNA expression of NOD1 and the p65 content in nuclear between 24-120 h(P<0.05),and the highest level at 48 h,subsequently the expression levels were began to decrease.The mRNA expression level of RIP2 was up-regulated after Hp was administrated, peaked at 48 h and declined after 72 h.However, the expression levels would rise again at 120 h.Conclusion: Hp infection can activate the NOD1/NF-κB signaling pathways and induce the production of IFN-βand IP-10 in gastrics of mice.

Objective To review the experience of colon interposition in the treatment of benign esophageal stricture.Methods 58 patients who had undergone colon interposition for esophageal replacements were studied retrospectively,including 53 patients with corrosive burn esophageal strictures,3 traumatic esophageal strictures and 2 congenital esophageal strictures.The interposition colon for all patients went through substemum paths.Results There was no postoperative death in the duration of hospital stay.14 cases developed postoperative complication including 2 total colon necrosis,7 anastomotic leak,2 anastomotic stricture and 3 recurrent laryngeal nerve injury.52 patients were followed-up(ranged 1 to 16 years),40 cases were extremely satisfied(1 grade),9 very satisfied(2 grade),2 satisfied(3 grade)and 1 unsatisfied(4 grade).Conclusion Colon interposition is an ideal procedure for esophageal benign stricture.

Objective To explore the appropriat.e treatment process for elderly patients with intertrochanteric fracture in primary hospitals.Methods According with the evaluation system and the characteristics of the grassroots hospitals and elderly patients with intertrochanteric fractures,the treatment standards for risk assessment was established,and the appropriate treatment process for intertrochanteric fracture in elderly patients in primary hospitals was initially formed and tried to promote applications in three primary hospital from December 2010 to January 2012,and its feasibility and effectiveness were tested.Results 66 elderly patients with intertrochanteric fracture were treated in three primary hospital,and 38 cases were hospitalized in primary hospital,including 24 cases of osseous union in 27cases treated with expectant treatment,11 cases of surgical treatment,and the cure rate was 89.5％.Sent on the operation rate of 90.3％ cases,54.8％ of cases occurred within 2 days.The incident of complications in early stage was 25.8％.Conclusion The diagnosis and treatment process and assessment criteria of treatment risk for elderly patients with intertrochanteric fracture in the primary hospital which is initial established is simple,practical,practicable,and has good effect and certain clinical value.

AIM: To study the basic mechanism of transcriptional regulation, NKX3.1 gene promoter was cloned and its promoter activities in prostate cancer cell lines and other cancer cell lines were tested. METHODS: 1.04 kb - promoter fragment of NKX3. 1 gene was obtained by PCR and cloned into pGL3 - basic and pEGFP - 1 that are promoter - less reporter vectors to examine its promoter activity driving the reporter gene transcription. The promoter activity was determined by dual -luciferase reporter assay and the expression of GFP reporter observed under fluorescence micro scope. RESULTS: The sequence of the cloned 1.04 kb promoter proved to be correct by DNA sequencing. The dual - lu ciferase reporter assay (M1/M2) showed that the promoter activity in LNCaP cell transfected with pGL3 - 1.04 kb promoter was about 1.5 - fold higher than that of pGL3 - control transfection and 50 - fold higher than that of pGL3 - basic transfec tion. To investigate the 1.04 kb - promoter activity in different tumor cell lines, the constructed pGL3 - 1.04 kb promoter and pEGFP - 1.04 kb promoter were transfected into several cell lines, respectively. The results showed that the activity of 1.04 kb promoterin LNCaP was highest among the tested cell lines. Multiple consensus sequence elements have been iden tified within the 1.04 kb fragment using TRANSFAC database. Further experiments will be done to determine their founc tions. CONCLUSION: Cloned 1.04 kb fragment upstream of NKX3.1 gene presented a strong promoter activity and its activity was highest in LNCaP cell among the tested tumor cell lines.

AIM: To observe the effect of exogenous androgen responsive element decoy on the promoter of prostate specific antigen (PSA) and the growth of LNCaP cells for searching the possibility of gene therapy for prostate cancer. METHODS: Firstly, pGL3 - PSA luciferase expression vector containing 640bp - promoter fragment of PSA gene was constructed. Then, a 23 -mer phosphorothioated ARE decoy based on the deduced ARE sequence at the promoter region of PSA gene was synthesized. pGL3 - PSA and ARE decoy DNA were cotransfected into PC3 - M cell by lipofectamineTM 2000. Through detecting the activity of luciferase, the effect of ARE decoy on the promoter of PSA was studied. Then the ARE decoy DNA was transfected into LNCaP cells. The effect of decoy DNA on the proliferation of LNCaP cells was examined by using MTT assay. The effects of apoptosis were detected by phase contrast microscopy, DNA agrose gel electrophoresis and flow cytometry. Meanwhile, the nuclear extract was prepared from LNCaP cells and DNA - protein interactions were examined by electrophoretic mobility shift assay (EMSA). RESULTS: The reporter assay showed that the aetivity of luciferase was significantly reduced in the ARE decoy - transfected cells, bnt not in the cells transfected with the control decoy. EMSA demonstrated specific binding of the ARE decoy to androgen receptor. The growth of LNCaP was remarkably inhibited and apoptotic morphological changes as well as DNA fragmentation were observed in the ARE decoy- transfected cells. The rate of apoptosis was 22.4% detected by FCM. CONCLUSION: The ARE decoy is capable of inhibiting the promoter of PSA gene and inducing the apoptosis in prostate cancer cells. It may become a potential therapeutic tool for prostate cancers.

Objective To investigate the clinical characteristics,the diagnosis and therapy of synchronous multiple primary esophageal carcinoma.Methods Thirty-two cases of synchronous multiple primary esophageal carcinoma were collected from January 1980 to December 2010 and their clinic data were retrospectively analysised.Results In the whole group,there were totally 66 lesions in 30 cases of double primary lesion and 2 cases of three primary lesion.The length of the lesions were 1 cm to 6.5 cm,and there were 22 lesions in cervical esophagus,10 in upper thoracic esophagus,19 in midthoracic esophagus and 15 in lower thoracic esophagus.Within the 66 lesions,65 lesions were squamous cell carcinoma and 1 was adenocarcinoma.The mucosa between the lesions were normal,with a distance of 4 cm to 9.5 cm,average 7.1 cm.Thirty-two patients with synchronous multiple primary esophageal carcinoma received surgery.Among 32 cases,26 of them were given definite diagnosis before operation.Four of them had exploratory operation,one patient underwent palliative resection,27 patients underwent radical resection.Two cases of the pathologic results of esophageal stump showed carcinoma after operation.The complications occurred in 8 patients.Twenty-eight cases were followed up after operation,the 1,3 and 5-survival rate were 76.9％,43.3 and 14.8％,respectively.Conclusion General pre-operation examination can significant helpful for the definite diagnosis of synchronous multiple primary esophageal carcinoma and surgical treatment is better choice for this disease.

AIM: To observe the effect of E2F decoy DNA on proliferation and apoptosis of androgen-independent prostate cancer cell line PC-3M.METHODS: E2F decoy DNA,ARE decoy DNA and control decoy DNA were transfected into PC-3M cells with lipofectamine,respectively.Their effects on cell proliferation were detected by MTT assay.The changes of cell morphology were observed by inverted phase contrast microscope.The cell apoptotic rate was determined by flow cytometry(FCM) analysis and chromosome DNA ladder was detected by DNA gel electrophoresis.The expression of c-Myc mRNA and cyclin D1 mRNA was detected by RT-PCR.The protein levels of c-Myc and cyclin D1 were detected by Western blotting.RESULTS: The growth of PC-3M cells was inhibited after transfection.The transfected PC-3M cells displayed typical apoptotic morphological changes.The apoptotic rate was 26.35% and DNA ladder was observed after transfection.The expression of c-Myc and cyclin D1 were inhibited.CONCLUSION: These results indicate that E2F decoy DNA induces apoptosis of androgen-independent prostate cancer cell lines PC-3M and inhibits cell proliferation via inhibiting expression of c-Myc and cyclin D1.

AIM: To study the basic mechanism of transcriptional regulation,NKX3.1 gene promoter was cloned and its promoter activities in prostate cancer cell lines and other cancer cell lines were tested.METHODS:(1.04 kb)-promoter fragment of NKX3.1 gene was obtained by PCR and cloned into pGL_3-basic and pEGFP-1 that are promoter-less reporter vectors to examine its promoter activity driving the reporter gene transcription.The promoter activity was determined by dual-luciferase reporter assay and the expression of GFP reporter observed under fluorescence microscope.RESULTS: The sequence of the cloned(1.04 kb) promoter proved to be correct by DNA sequencing.The dual-luciferase reporter assay(M1/M2) showed that the promoter activity in LNCaP cell transfected with pGL_3-(1.04 kb) promoter was about 1.5-fold higher than that of pGL_3-control transfection and 50-fold higher than that of pGL_3-basic transfection.To investigate the 1.04 kb-promoter activity in different tumor cell lines,the constructed pGL_3-(1.04 kb) promoter and pEGFP-1.04 kb promoter were transfected into several cell lines,respectively.The results showed that the activity of(1.04 kb) promoter in LNCaP was highest among the tested cell lines.Multiple consensus sequence elements have been identified within the(1.04 kb) fragment using TRANSFAC database.Further experiments will be done to determine their founctions.CONCLUSION: Cloned 1.04 kb fragment upstream of NKX3.1 gene presented a strong promoter activity and its activity was highest in LNCaP cell among the tested tumor cell lines.

To explore the relationship and possible mechanism of reproductive function of bisphenol A (BPA) with polycystic ovary syndrome (PCOS).Serum specimens were collected from 43 PCOS patients and 37 healthy women.And the level of BPA was detected by enzyme-linked immunosorbent assay (ELISA).The serum level of BPA in PCOS were significantly higher than that in the control group.And the inter-group difference was significant (P ＜ 0.05).The patient level of BPA was positively related with obesity and basic luteinizing hormone (LH) (P ＜ 0.05).In the PCOS group,no significant correlation existed between hyperandrogenemia and non-hyperandrogenemia (P ＞ 0.05).Although no difference existed between insulin and non-insulin resistance,the level of BPA in insulin resistance group was lower than in non-insulin resistance group.A higher level of BPA may be associated with obesity and basic LH in PCOS.