Objective To construct the eukaryotic expression plasmid containing glyceraldehydes-3-phosphate dehydrogenase (GAPDH) and cysteine protease inhibitor ( CPI ) gene from periodic Brugia malayi (Bm) and to lay foundation for studying multivalent vaccines. Methods Total RNA was extracted from periodic Bin. The BmGAPDH and BmCPI genes were amplified by RT-PCR. The PCR product was cloned and then subeloned into eukaryotic recombinant plasmid vector pcDNA3.1 (+). pcDNA3.1 (+)/BmGAPDH/BmCPI was constructed. The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification, and were transformed into HeLa cell subsequently. The transient expression of BmGAPDH and BmCPI were examined by RT-PCR. The expressed protein was identified by sodium dodeeylsulphate-polyacrylamide gel electrophoresis(SDS-PAGE). Results Two specific bands of around 877 bp of BmGAPDH and 621 bp of BmCPI were amplified, consistent with the expected value. The same bands were obtained by double restriction enzyme digestion of recombinant plasmids or PCR using recombinant plasmid as template. BmGAPDH and BmCPI mRNA were highly expressed in transfeeted HeLa cell. The relative molecular mass (Mr) of the recombinant protein was about 54 × 103. Conclusion The recombinant eukaryotic expression plasmid pcDNA3.1 (+)/BmGAPDH/BmCPI has been constructed successfully and the protein is expressed correctly in mammalian cell.

Objective To clone and sequence the cysteine protease inhibitor gene of periodic Brugia malayi(BmCPI) and predict B-cell epitopes in amino acide sequence of BmCPI in order to provide basis for further study the expression of BmCPI and its function. Methods Total RNA was extracted from periodic Brugia malayi.A couple of specific primers were designed on the basis of known sequences of cysteine protease inhibitor gene from BmCPI. The desired gene was amplified by PCR technique from cDNA. The PCR products were purified and cloned into plasmid pGEM-T by T-A cloning method, transformed into Escherichia coli(E, coli) strain DH5α. The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification. Five parameters and methods were used to predict B-cell epitopes in amino acide sequence of BmCPI. Results For RT-PCR, a specific band of around 621 bp was amplified. The same band was obtained by double restriction of recombinant plasmids or PCR using recombinant plasmid as template. The result of DNA sequencing showed that BmCPI shares 99% nucleotide sequence identity with that of published sequence. It showed that B-cell epitopes were probably at or adjacent to 23 - 32, 50 - 79 and 117 - 126 in its amino acide sequence. Conclusions pGEM-BmCPI is successfully constructed and sequenced, anticipated objective is reached and conditions is provided for further study of BmCPI expression and its function.

OBJECTIVETo study the relationships of hypoxia-induced factor (HIF) with tumor angiogenesis and early liver metastasis in colonic cancer.

METHODSThirty three cases of colon cancer undergoing radical surgery were divided into two groups according to liver metastasis or not within half a year after operation. Expressions of HIF and vascular endothelial growth factor (VEGF) were examined using immunohistochemical method and tumor microvessel density (MVD) was measured in colonic cancer specimens.

RESULTSFifteen cases developed early liver metastasis, while 18 did not. The positive rates of HIF and VEGF, and MVD were 86.7%, 66.7%, (57.9+/- 12.7)% respectively in the group with early liver metastasis, significantly higher than 44.4% (P< 0.05), 27.8% (P< 0.05) and (22.3+/- 10.2)% (P< 0.01) respectively in the group without early liver metastasis respectively.

CONCLUSIONHIF can promote tumor angiogenesis in colonic cancer, and is closely related with early liver metastasis.

Adult ; Aged ; Colonic Neoplasms ; metabolism ; pathology ; Female ; Follow-Up Studies ; Humans ; Hypoxia-Inducible Factor 1 ; metabolism ; Liver Neoplasms ; blood supply ; secondary ; Male ; Microvessels ; Middle Aged ; Neovascularization, Pathologic ; Vascular Endothelial Growth Factor A ; metabolism

The chitosan thermosensitive hydrogel is liquid at room temperature but gels rapidly when heated to body temperature. This hydrogel are wildly used for cell encapsulation, drug delivery or tissue-engineered scaffolds. The system can sustain the release of macromolecules over a period of several hours to a few days. However, with low-molecular-weight compounds, the release is generally completed within 24 h. To prepare a functional chitosan thermosensitive hydrogel for slow release both broad-spectrum antibiotic chlorhexidine and growth factor recombined human bone morphogenetic protein-2 (rhBMP-2), The beta-cyclodextrin was used to prepare an inclusion complex with chlorhexidine, and then the latter was incorporated into the chitosan thermosensitive hydrogel system. Simultaneously, rhBMP-2 was added into the hydrogel system. By HAAKE viscosity measuring instrument, we contrasted the viscoelastic properties of system with or without objective factors. And the in vitro release kinetics of chlorhexidine and rhBMP-2 was investigated by HPLC (high performance liquid chromatography) and ELISA (enzyme-linked immunosorbent assay) respectively. The results showed that the addition of chlorhexidine/beta-cyclodextrin inclusion complex to the thermosensitive solution did not change the gelling behavior of the thermosensitive system. Further, the in vitro release profiles demonstrated that the release rate of chlorhexidine and rhBMP-2 from hydrogel became slower, controlled delivery over at least 1 month. By first preparing chlorhexidine/beta-cyclodextrin inclusion complex, and then mixing the IC and rhBMP-2 into the chitosan thermosensitive hydrogel, a functional chitosan thermosensitive hydrogel system with ability of slow release both rhBMP-2 and chlorhexidine is successfully made.
Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; administration & dosage ; Chitosan ; chemistry ; Chlorhexidine ; administration & dosage ; Delayed-Action Preparations ; chemical synthesis ; Drug Carriers ; chemistry ; Drug Combinations ; Hydrogels ; chemistry ; Recombinant Proteins ; administration & dosage ; Temperature ; Transforming Growth Factor beta ; administration & dosage

OBJECTIVETo investigate the clinicopathological features, diagnosis, treatment and prognosis of primary clear cell carcinoma of the liver (PCCCL).

METHODSThe clinicopathological data of 24 cases with pathologically proven PCCCL in the First Affiliated Hospital of Guangxi Medical University from May 1996 to December 2003 were collected and analyzed.

RESULTSThere were 21 males and 3 females in this group, with an average age of 46 years (range: 30 approximately 78 years). HBV infection was detected in 83.3%, and AFP expression was found in 75.0% of them. Of the 24 cases, 28 tumors were found with an average size of (6.64 +/- 5.54) cm. Liver cirrhosis was found in 75.0% of the patients. Macroscopic and microscopic tumor thrombi were found in 20.8% and 29.2%, respectively. Lymph node metastasis was found in 4.2% of the patents. The 1-, 3-, and 5-year overall survival rates of the 24 cases were 75.0%, 41.7% and 27.8%, respectively, with a median survival time of 29 months.

CONCLUSIONThe clinical characteristics of primary clear cell carcinoma of the liver are similar to that of common hepatocellular carcinoma. It is difficult to be diagnosed preoperatively and final diagnosis depends on pathological examination. Surgical resection is an effective way to achieve favorable treatment outcome and even long-term survival.

Adenocarcinoma, Clear Cell ; blood ; pathology ; surgery ; virology ; Adult ; Aged ; Female ; Follow-Up Studies ; Hepatectomy ; methods ; Hepatitis B ; Humans ; Liver Neoplasms ; blood ; pathology ; surgery ; virology ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Survival Rate ; alpha-Fetoproteins ; analysis

OBJECTIVETo study the optimal surgical resection length for esophageal carcinoma.

METHODSSpecimens of seventy patients with esophageal squamous cell carcinoma resected and collected in our hospital were made into pathologic giant sections. Direct intramural infiltration, multicentric carcinogenic lesion and leaping metastasis were observed in the large slice by microscope. The actual length during the operation was calculated by the ratio of shrinkage.

RESULTSDirect intramural infiltration was found in 51 (72.9%) patients, 39 proximal and 36 distal to the tumor. The mean length of direct intramural infiltration was 0.9 +/- 0.8 cm (4.0 cm maximum) proximally and 0.5 +/- 0.3 cm (2.0 cm maximum) distally. Multicentric carcinogenic lesion was found in 11 (15.7%) patients, 5 proximally, 8 distally and 2 on both sides. Proximal to the tumor, the mean distance between the multicentric carcinogenic lesion and the main lesion plus the length of the multiple carcinogenic lesion was 3.2 +/- 1.5 cm (4.7 cm maximum). Distal to the tumor, it was 3.6 +/- 2.4 cm (9.1 cm maximum). Leaping metastasis was found in 9 (12.9%) patients, 7 proximally and 4 distally. The mean distance between the leaping metastasis and the main lesion plus the length of the leaping metastatic lesion was 1.9 +/- 0.6 cm (2.9 cm maximum) proximally and 1.4 +/- 1.0 cm (2.7 cm in maximum) distally.

CONCLUSIONThe optimal surgical resection length for esophageal carcinoma should be at least 5 cm proximal to the tumor and total length on the distal side.

Esophageal Neoplasms ; pathology ; surgery ; Female ; Humans ; Male ; Neoplasm Invasiveness

According to the 2020 world cancer data report， the prevalence of prostate cancer ranks second in male malignant tumors and the mortality fifth. In China， due to the poor living and eating habits， the prevalence of prostate cancer is still rising， and advanced prostate cancer can adapt to and tolerate androgen castration related drugs through a variety of mechanisms， which often indicates poor therapeutic outcomes in the late stage. Houttuyniae Herba， a medicinal plant of family Saururaceae， has been officially identified both as food and medicine by the National Health Commission. The existing studies have shown that Houttuyniae Herba inhibits the proliferation， migration， and invasion of prostate cancer and induces its apoptosis and cell cycle arrest via multiple targets， links， and pathways. These anti-tumor activities can also be observed in breast cancer， leukemia， gastric cancer， colorectal cancer， and other tumor cells. Such activities are mainly related to the inhibited expression of cyclin and cyclin-dependent kinase， regulation of apoptosis-related proteins， activation of apoptosis-related pathways， reduced secretion of matrix metalloproteinase， inhibition of activator of transcription 3 （STAT3）/Zinc finger protein （Snail） / transcription factor twist signaling pathway， and antagonization of growth factor receptor， androgen， inflammation， and oxidative stress in promoting the occurrence and development of prostate cancer. In addition， Houttuyniae Herba is able to alleviate diabetes， regulate blood glucose-lowering pathways， and boost immunity by inhibiting the protein kinase B （Akt）/mammalian target of rapamycin （mTOR） and mitogen-activated protein kinase （MAPK） signaling pathways， thus suppressing the occurrence and progression of prostatic cancer. In conclusion，Houttuyniae Herba can be used as a potential drug against prostate cancer.

OBJECTIVETo explore the potential risk factors of intrahepatic cholangiocarcinoma (ICC) in China.

METHODA case-control study including 317 patients with pathologically confirmed ICC and 634 healthy individuals was conducted. The cases and controls were matched in age, sex and inhabitancy. Data were statistically analyzed by Chi-square test and conditional logistic regression.

RESULTSUnivariate analysis showed significant difference in HBsAg seropositivity, liver cirrhosis, hepatolithiasis, choledocholithiasis and schistosomiasis between ICC patients and healthy controls (P < 0.05). Multivariate analysis confirmed that HBsAg seropositivity, liver cirrhosis, hepatolithiasis and hepatic schistosomiasis were associated with ICC, and their adjusted odds ratio (95% confidence interval) were 10.265 (6.676-15.783), 13.101 (5.265-32.604), 18.242 (3.580-92.958), 18.435 (1.930-176.082), 15.102 (4.607-49.499) and 11.820 (3.522-39.668), respectively. The incidence of hepatic cyst, cholecystolithiasis, hepatic hemangioma, fatty liver, diabetes mellitus, smoking and drinking were not significantly different between ICC patients and controls.

CONCLUSIONSThe HBV infection, liver cirrhosis, hepatolithiasis and hepatic schistosomiasis may be the risk factors for ICC in China.

Adult ; Aged ; Bile Duct Neoplasms ; epidemiology ; etiology ; Bile Ducts, Intrahepatic ; Case-Control Studies ; Cholangiocarcinoma ; epidemiology ; etiology ; Cholelithiasis ; complications ; epidemiology ; Female ; Hepatitis B ; complications ; epidemiology ; Hepatitis B Surface Antigens ; blood ; Hepatitis B e Antigens ; blood ; Humans ; Liver Cirrhosis ; complications ; epidemiology ; Liver Diseases ; complications ; epidemiology ; Logistic Models ; Male ; Middle Aged ; Odds Ratio ; Risk Factors

BACKGROUND AND OBJECTIVEMultinodular hepatocellular carcinoma(HCC) might originate from multicentric occurrence (MO) or intrahepatic metastasis(IM). This study was to find out proteins which play important roles in clonal origin of multinodular hepatocellular carcinoma bt screening the differentially expressed proteins between the MO and IM tissues using comparative proteomic analysis.

METHODSTotal protein extracted was separated by two-dimensional gel electrophoresis. Comparative analyses of the 2-DE protein patterns between the two groups were carried out using computerized imaging techniques. Proteins exhibiting significant alternations were subsequently isolated and identified by mass spectrometry.

RESULTSA total 1025+/-52 and 900+/-98 spots were detected in the protein profile in IM and MO, respectively. Twenty-five protein spots were statistically different at expression levels between the two groups. Twenty of them were identified by MALDI-TOF-MS and bioinformatics.

CONCLUSIONSThe protein profile of MO HCC tissues is different from that in IM HCC tissues. The twenty differentially expressed proteins might play a key role in the carcinogenesis and progression of multinodular HCC. These newly identified proteins might be potential and valuable biomarkers for identifying the multinodular HCC of clonal origin.

Adult ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Electrophoresis, Gel, Two-Dimensional ; Female ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Metastasis ; Neoplasms, Multiple Primary ; metabolism ; pathology ; Protein Array Analysis ; Proteomics ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Objective?To evaluate the application of guidewire-assisted biopsy and balloon dilatation cytology smear through ERCP in diagnosis of patients with hilar cholangiocarcinoma.?Methods?During the ERCP procedure, 52 patients with hilar cholangiocarcinoma were treated with balloon dilatation, and cytology smear from the surface of the balloon. At the same time, biopsy forceps assisted by guidewire inserted into the bile duct to biopsy the lesion and obtain specimens.?Results?Success rate of obtaining histopathological through ERCP guidewire assisted biopsy in biliary bile duct was 100.0%, 31 cases were histological diagnosed by forceps biopsy in 52 cases, positive diagnosis rate was 59.6%; The patient with balloon dilatation in 52 cases give cytology smears obtained diagnosis in 22 cases, the positive rate was 42.3%; cytology smear combined with biopsy, a total of 34 cases were diagnosed, the positive rate was 65.4%. There was no serious complications occurred.?Conclusion?Biopsy assisted by guidewire through ERCP is a safe, simple, easy technique and an effective means of obtaining pathological, meanwhile, the cytology smear after balloon dilatation can be used as a supplement to biopsy, and provides an effective means in pathological diagnosis of hilar cholangiocarcinoma.