Porphyromonas gingivalis is a specific causative agent of human chronic periodontitis. This anaerobe produce collagenase PrtC encoded by prtC. To constructed the prokaryotic expression system for the prtC gene from P.gingivalis so as to be used to explore antigenicity and immunoreactivity of prtC gene product as well as the relationship between the local level of PrtC and the periodontitis damage, the entire length of prtC genes fragment from the ATCC-33277 and 47A-1 strains of P.gingivalis was amplified by PCR. After T-A cloning and sequencing, the prokaryotic expression system for prtC was constructed by using pET32a plasmid and E.coli BL21DE3 strain. The expression of the target recombinant PrtC protein was induce with different concentrations of IPTG. Western blot assay was used to detect the antigenicity and immunoreactivity of PrtC protein, and ELISA assay was used to detect the PrtC level in the subgingival plaque samples of patients with chronic periodontitis. The experimental results showed that the nucleotide sequences of the prtC genes from ATCC-33277 and 47A-1 strains of P.gingivalis were entirely identical, and the sequence similarities of nucleotides and amino acids were 98.46% and 99.07% respectively. Under the induction of different concentration of IPTG, the output of recombinant expressed product PrtC may reach up to 50% of the total bacterial proteins. It was also proved that the recombinant PrtC could bind with antibody against the whole cell of P.gingivalis, and could induce the production of specific antibodies in rabbit. In 91.39% of the subgingival plaque samples, the PrtC could be detectable, in which the level of positive rates of detection was higher in severe cases of chronic periodontitis than that in the mild cases. So far, a prokaryotic expression system for the prtC gene from P.gingivalis with high expression efficiency was successfully constructed in the present study, and the expressed product PrtC possesses well antigenicity and immunoreactivity, suggesting the possibility to be used as the candidate antigen for developing the serological kit and P.gingivalis vaccine.

OBJECTIVE:To emphasis the importance to unceasingly upgrading of drug quality standard for drug quality and to arouse the attention of the related enterprises, medical institutions and governmental department to this matter. METHODS: The correlation between quality standard and drug quality was expounded systematically by citing examples and reviewing the pertinent literature, and the current status of drug quality standard in China was analyzed briefly. RESULT & CONCLUSION: Great importance should be attached to the evaluation of drug quality standard to improve drug quality standard level step by step and to ensure efficacy and safety of drugs.

OBJECTIVE:To study the effects of modern decoction method vs. traditional decoction method on decoction rate and decoction quality. METHODS: The advantages and disadvantages were compared between the two decoction methods through analysis on the whole decoction process and the storage of the decoction before oral administration taking major components or active components as parameters. RESULTS: As compare with traditional decoction method, the modern decoction method had more advantages for in which the operation is standard, the contents of active components were high, and the quality control can be performed from many ways. CONCLUSION: The modern decoction method deserves to be popularized.

Objective To construct a recombinant adenovirus vector encoding for indoleamine 2,3-dioxygenase(IDO)and chimetric albumin promoter,evaluate the mRNA and protein expression levels in Hepa 1-6cell.Methods Full-length mouse derived IDO cDNA was subeloned into pAdTraek-ALB shuttle Plasmid.The product was linearized to homologous recombination with AdEasy-l vector in BJ5183 bacteria.The positive clone was identified by restriction endonuclease digestion and further confirmed by sequencing.The recombined adenoviruses DNA were transfected into AD-293 cells for packaging and amplification of Ad-ALB/IDO.The expression of IDO was monitored by RT-PCR and EGFP fluorescence in infected cells.The recombinant viruses with Hepa 1-6 cells were cultured and the mRNA and protein expression levels monitored bv RT-PCR and Western blot, respectively. Results Construction of recombinant andenoviruses containing IDO and albumin promoter was confirmed by restriction endonuclease digestion and sequencing.The expression of IDO was identified by RT-PCR in transfected AD-293 cell.The virus titer was 2.9×10~6 pfu/ml.The IDO mRNA and protein expression levels were detectable after transfected Hepa 1-6 cells by RT-PCR and Western blot. Conclusion A recombinant adenovirus Ad-ALB/IDO was susceessfullyconstructed.

Objective To investigate the effectiveness and technical points of anatomical liver resection by trans-Glisson Sheath methylene blue staining in treatment of hepatolithiasis of right posterior lobe.Methods The clinical data of 12 cases of hepatolithiasis of right posterior lobe treated with anatomical liver resection by Glisson sheath methylene blue staining were retrospectively analyzed.Result 6 of 12 patients had undergone more than 2 previous biliary surgeries.All patients underwent contrast-enhanced CT scan and portography,hepatolithiasis of segment Ⅵ in 4 cases,right posterior lobe in 8 cases,accompanied by left lateral lobe bile duct stones in 2 cases,the right caudate lobe bile duct stones in 1 case.Methylene blue was injected into the portal vein,the methylene blue interface of segment Ⅵ or right posterior lobe displays well.Methylene blue interface was larger than the ischemia interface,which is in accordance with the anatomy.Along the methylene blue interface,hepatic resection was performed including right posterior lobe resection (n =9),segment Ⅵ resection (n =3),and combined with left lateral lobe resection (n =2) and the right caudate lobe resection (n =1).There was no postoperative mortality.Incision infection occurred in 5 cases,4 had right pleural effusion and 2 had a biliary fistula that were treated conservatively.With a mean follow-up period of 3.2 years,all patients are symptoms free and stone free.Conclusions Anatomical liver resection by methylene blue staining is a safe and effective treatment for hepatolithiasis of right posterior lobe.

Objective To investigate the clinicopathologic features and prognostic analysis of intrahepatic mucinous cholangiocarcinoma (IMCC) and intrahepatic cholangiocarcinoma (ICC).Methods The retrospective cohort study was adopted.The clinicopathological data of 20 patients with IMCC and 51 patients with ICC who were admitted to the Fujian Provincial Hospital from March 2003 to March 2014 were collected.The observation indicators included (1) clinicopathologic features:gender,age,clinical symptoms (fever,jaundice,abdominal pain and peritoneal effusion),preoperative laboratory examination [alanine transaminase (ALT),aspartate transaminase (AST),direct bilirubin (DBil),carcinoembryonic antigen (CEA),CA19-9 and alphafetoprotein (AFP)],operation methods (radical resection,palliative resection) and pathological features (tumor location,lymph node metastasis,vascular invasion and pathological stage).(2) The follow-up of outpatient examination and telephone interview was performed to detect the survival of patients up to March 2015.Measurement data with normal distribution were represented as x-± s and comparison between groups was analyzed using t test.Count data were analyzed using the chi-square test.The survival curve was drawn by Kaplan-Meier method,and the survival rate was analyzed using the Log-rank test.Result The numbers of patients with fever,jaundice,elevated level of AST and DBil,lymph node metastasis,vascular invasion and numbers of patients with stage 0,Ⅰ,Ⅱ,Ⅲ,Ⅳ were 13,11,10,11,3,1 and4,6,2,6,2 with IMCC,12,10,9,2,22,36 and 0,14,5,3,29 with ICC,respectively,showing statistically significant differences in the above indicators (x2=10.830,8.639,7.672,25.059,8.036,24.765,26.601,P ＜ 0.05).All the patients were followed up for a median time of 14 months (range,1-118 months).The survival time and 1-,3-,5-year survival rates were (55 ± 8)months,94.4％,44.0％,16.7％ in 20 patients with IMCC and (30 ±6) months,36.5％,12.5％,4.0％ in 51 patients with ICC,respectively,showing a statistically significant difference (x2 =8.126,P ＜ 0.05).Conclusion The patients with MICC are more easily complicated with fever,jaundice and liver dysfunction,while they have less lymph node metastasis and vascular invasion,earlier pathological stage and better prognosis compared with patients with ICC.

Familial adenomatous polyposis is characterized by the multiple and adenomatous polyps in the colorectum combined with polyps in the stomach and duodenum, while it is rarely seen in the common bile duct (CBD).In July 2013, 1patient with FAP combined with adenomas in the CBD was admitted to the Fujian Provincial Hospital.The patient underwent laparoscopic CBD exploration and resection of masses due to acute pancreatitis 11 months ago, and was confirmed as with adenoma in the distal CBD by postoperative pathological examination.Multiple polyps were found in the stomach, duodenum,CBD, colorectum after admission to hospital, biopsy confirmed that polyps were tubular adenoma.The patient received pancreaticduodenectomy and was diagnosed as with duodenum-CBD tubular adenoma in postoperative pathological examination.The patient was followed up by telephone interview and outpatient examination and had a full recovery, in addition to the increasing of stools frequency and occasioned hematochezia, and then was treated by total colectomy at postoperative month 7.The tubular adenoma was confirmed by postoperative pathological examination.

Objective:To analyze the feature of anaphylactic shock induced by alprostadil injection. Methods: Totally 1624 pa-tients treated with alprostadil injection in our hospital from January to March 2014 were undergone adverse drug reaction monitoring. Six cases of anaphylactic shock induced by alprostadil injection were statistically analyzed. Results:6 cases of anaphylactic shock induced by alprostadil injection occurred in the department of intervention and most of thd reaction happened in arterial injection. Two lot num-ber of drugs were in high frequency. Patients showed a sharp fall in blood pressure, chest discomfor, palpitation, shortness of breath, paleness, sweating and other symptoms. After anti allergy, blood pressure, oxygen the rapy and other treatment measures vital signs gradully stabilized. Conclusion:Anaphylactic shock induced by alprostadil injection is in correlation with the administration mode of artery injection. The change of the patient' s condition and vital signs should be closely observed when we inject the patient with alpros-tadil injection. We must prevent the occurrence of serious adverse reactions.

Objective To investigate the diagnosis and treatment of mucin-producing intrahepatic biliary tumor (MPIBT).Methods We retrospectively analyzed the clinical,radiologic,surgical and pathologic findings of 16 MPIBT cases from January 2004 to December 2011.Results There were six men and ten women,age ranged from 44 to 69 years (mean 60 years).Clinical presentation included jaundice with abdominal dull pain in 5 patients,acute cholangitis in 4 patients,painless jaundice in 2 patients,upper abdominal dull pain in 3 patients,no obvious symptoms in 1 patient,body weight loss more than 5 kg within 3 months in 5 patients.The most characteristic appearance of MPIBT on magnetic resonance cholangiopancreatography were asymmetry of intrahepatic bile duct dilatation and the dilatation in both extraand intrahepatic bile duct distal to the hepatic mass and not sudden interruption in extrahepatic bile duct.The primary tumor located in the left hepatic bile duct in 15 cases,in the right hepatic bile duct in one.13 MPIBT cases received hemihepatectomy and extrahepatic bile duct resection and Roux-en-Y anastomosis was done in 8 cases,3 received palliative biliary drainage.Pathologically 13 was papillary adenocarcinoma and 3 was papillary adenoma.The 1-,2-,3-year survival rates for the 16 MPIBT patients were 81％,66％,56％,respectively.Conclusions MPIBT had no specific clinical manifestations,MRCP might be an effective means for the diagnositic strategy and assessment of tumor extension before surgery,radical resection was the first choice of treatment,palliative biliary drainage could prolong the survival time.

Objective To investigate kupffer cells (KCs) expressing indoleamine 2,3-dioxygenase(IDO)in the inhibition of allogeneic T-cell proliferation in vitro. Methods Real-time PCR was used to investigate the expression of IDO mRNA and FasL mRNA in KCs pretreated with or without IFNγ. High performance liquid chromatography was used to analyze the catabolism of tryptophan by IDO from KCs. Allogeneic T-cell response was used to confirm the inhibition of KCs in vitro. The proliferation of lymphocytes was detected using [3 H] thymidine incorporation. Cell cycle and lymphocyte apoptosis were evaluated by flow cytometric assay. Results Real-time PCR revealed IDO mRNA and FasL mRNA expression in KCs pretreated with IFN-γ. IDO catabolic effect was confirmed by a decrease in tryptophan and increase in kynurenine concentration. KCs expressing IDO and FasL from BABL/c mice acquire the ability to suppress the proliferation of T-cells from C57BL/6, which could be blocked by the addition of 1-methyl-tryptophan and anti-FasL antibody. The co-cultured T-cells with KCs expressing IDO and FasL could induce allogeneic T-cell apoptosis and exhibited cell-cycle arrest in G1. Conclusion In addition to the Fas/FasL pathway, IDO may also play an important role in KCs to inhibit allogeneic T-cell proliferation in vitro.