OBJECTIVETo analyze the proteomic components of the sera from the patients with hepatocellular carcinomas (HCC), in search of the diagnostic markers of HCC.

METHODSImmobilized pH gradient two-dimensional polyacrylamide gel electrophoresis (2DE), silver staining, PDQuest 2DE analysis software, peptide mass fingerprint based on matrix-assisted laser desorption/ionization time of flying mass spectrometry (MALDI-TOF-MS) and SWISS-PROT database searching, were used to separate and identify the proteome of the sera from the patients with HCC.

RESULTSThe 2DE images were analysed by PDQuest 2DE software. The average matching rate was 70.2%. In IEF direction, the average deviation was (1.02 +/- 0.22) mm, in SDS-PAGE direction, the deviation was (0.97 +/- 0.14) mm. The twenty-three different protein spots were incised from silver staining gel and digested in-gel by TPCK trypsin. 15 peptide mass fingerprints (PMF) maps were obtained by MOLDI-TOF-MS. The typical peptide masses were searched in the SWISS-PROT database using PeptIdent software.

CONCLUSIONSGood reproducibility could be obtained by applying immobilized pH gradient two-dimensional polyacrylamide gel electrophoresis (2DE) to separate and identify the proteome in serum. There is still the problem of efficiently removing a higher level of proteins and lipids from the serum. Identification by MOLDI-TOF-MS peptide mass fingerprint provides useful information for screening diagnostic markers of human HCC.

Blood Protein Electrophoresis ; Blood Proteins ; analysis ; Carcinoma, Hepatocellular ; blood ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Liver Neoplasms ; blood ; Proteome ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Objective To investigate the risk factors for malignant transformation of oral leukoplakia. Methods A total of 409 cases with oral leukoplakia was retrospectively analyzed. Single factor test was first performed to examine the associations between oral leukoplakia's histopathological classification and each of risk factors including sex, age, systemic diseases, course of disease, clinical classification,site, size, numbers of lesion, alcohol and tobacco consumption, and symptom. Then the association of these selected factors with oral leukoplakia's histopathological classification was evaluated using multiple logistic regression analysis. Results Fifty-two cases of all 409 patients with oral leukoplakia (including 9 severe dysplasia) developed oral cancer. The ratio of malignant transformation was 12.7%. Sex, age, clinical type, site and symptom were chosen as risk factors incorporated into the multiple logistic regression models.The risk of mild-moderate dysplasia in female was 2. 40 times as high as that in male. The risk of mild-moderate dysplasia of speckled leukoplakia was 2. 81 times as high as that of homogeneous leukoplakia. The risk of mild-moderate dysplasia of dangerous site was 1. 98 times as high as that non-dangerous site. The risk of mild-moderate dysplasia with symptom was 1. 84 times as high as that without symptom. The risk of severe dysplasia and oral cancer in female was 3. 11 times as high as that in male. The risk of severe dysplasia and oral cancer of speckled (4. 50 times), ulcerative (5. 63 times), verrucous leukoplakia (4.09 times) were much higher than that of homogeneous leukoplakia. The risk of severe dysplasia and oral cancer in dangerous site was 2. 79 times as high as in non-dangerous site. The risk of severe dysplasia and oral cancer in leukoplakia with symptom was 4. 38 times as high as without symptom. Conclusions The malignant transformation of oral leukoplakia is correlated to sex, clinical type, site and symptom.

OBJECTIVEThe pulmonary toxicity of water extract of Siegesbeckia pubescens has been studied an mice following subchronic oral administration route.

METHODMice were randomly grouped and administered with the water extract of S. pubescens at dosages of 3.0, 1.0, 0.3 g x kg(-1) and saline respectively. 10 mice of each group were sacrificed on the day of 7 th, 14 th, 21 th, and 2 weeks after stopping administration, histological changes of the lung were examined.

RESULTThe water extract of S. pubescens at dosage of 3.0 g x kg(-1) increased the lung index on the of day 14 th and 21 th, significant histopathological damages were observed. The histopathological changes were disappeared after stopping administration for 2 weeks.

CONCLUSIONThe water extract of S. pubescens has a pulmonary toxic effect on mice, and the toxic effect is reversible.

Administration, Oral ; Animals ; Asteraceae ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; toxicity ; Female ; Lung ; drug effects ; Male ; Mice ; Random Allocation

BCMA is one of the transmembrane receptors belonging to BAFF and APRIL. In order to identify the feasibility of sBCMA as decoy receptor and obtain active sBCMA for its structural and functional research, full length of hBCMA was amplified with total RNA from Raji cell line by RT-PCR, and the cDNA encoding the extracelluar soluble domain of hBCMA was inserted into pET43.1a(+) vector. The recombinant vector pET43.1a(+)-sBCMA was transformed into E. coli Origami B(DE3) pLyS which is helpful for disulfide bond construction of expression proteins. After IPTG induction, the recombinant protein was expressed as soluble fusion protein, sBCMA-NusA-His6, and identified by western blotting. Then the target protein was purified by Ni(+)-chelating Sepharose Fast Flow. The binding activity between recombinant sBCMA and BAFF was detected by ELISA. Also, Recombinant sBCMA inhibited proliferation of mouse B cell stimulating by rhsBAFF. It was proved that recombinant sBCMA has good bioactivity and the method to express those proteins rich in disulfide bond is feasible and effectual.
B-Cell Activating Factor ; chemistry ; B-Cell Maturation Antigen ; biosynthesis ; genetics ; Cloning, Molecular ; DNA, Complementary ; genetics ; Disulfides ; chemistry ; Escherichia coli ; genetics ; metabolism ; Humans ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Solubility ; Tumor Necrosis Factor Ligand Superfamily Member 13 ; chemistry

BACKGROUNDZengshengping (ZSP) tablets had inhibitory effects on oral precancerous lesions by reducing the incidence of oral cancer. However, the severe liver toxicity caused by systemic administration of ZSP limits the long-term use of this anti-cancer drug. The purpose of this study was to evaluate the tumor inhibitory effects due to the topical application of extracts from ZSP, a Chinese herbal drug, on 7, 12-dimethlbenz(a)anthracene (DMBA) induced oral tumors in hamsters. The study also investigated the anti-cancer mechanisms of the ZSP extracts on oral carcinogenesis.

METHODSDMBA (0.5%) was applied topically to the buccal pouches of Syrian golden hamsters (6 - 8 weeks old) three times per week for six weeks in order to induce the development of oral tumors. Different fractions of ZSP were either applied topically to the oral tumor lesions or fed orally at varying dosages to animals with oral tumors for 18 weeks. Tumor volume was measured by histopathological examination. Tumor cell proliferation was evaluated by counting BrdU labeled cells and by Western blotting for mitogen-activated protein kinase (MAPK) protein levels. The protein levels of apoptosis marker Caspase-3 and regulator Bcl-2 protein were also measured by Western blotting.

RESULTSTopical application of DMBA to the left pouch of hamsters induced oral tumor formation. Animals treated with DMBA showed a loss in body weight while animals treated with ZSP maintained normal body weights. Both the ZSP n-butanol fraction and water fraction significantly reduced tumor volume by 32.6% (P < 0.01) and 22.9% (P < 0.01) respectively. Topical application of ZSP also markedly decreased the BrdU-positive cell numbers in oral tumor lesions and reduced the expression level of MAPK. In addition, ZSP promoted tumor cell apoptosis by increasing Caspase-3 expression but decreasing Bcl-2 protein production.

CONCLUSIONThe n-butanol and water fractions of ZSP are effective at inhibiting tumor cell proliferation and stimulating apoptosis in oral cancer suggesting that these fractions have chemopreventive effects on DMBA induced oral carcinogenesis.

9,10-Dimethyl-1,2-benzanthracene ; toxicity ; Animals ; Antineoplastic Agents ; therapeutic use ; Cell Transformation, Neoplastic ; drug effects ; Cricetinae ; Drugs, Chinese Herbal ; therapeutic use ; Male ; Mesocricetus ; Mouth Neoplasms ; chemically induced ; drug therapy ; prevention & control

The characteristics of purinoceptors in the membrane of rat trigeminal ganglion (TG) neurons were studied by using whole- cell patch clamp technique. The results showed that most of neurons examined (78.9%, 142/180) were responsive to ATP in a concentration-dependent manner; the others (21.1%, 38/180) were ATP insensitive. Of the ATP-sensitive cells, the majority (95.1%, 135/142) responded to ATP with an inward current, a few (2.1%, 3/142) with an outward current, and the rest (2.8%, 4/142) with biphasic current. Small sized cells (<30 mum) responded to ATP with a rapid desensitizing inward current and were highly sensitive to vanilloid; the medium sized cells (30~40 mum) responded to ATP with slow desensitizing inward current and were not sensitive to vanilloid; while the majority of large sized cells (>40 mum) did not respond to ATP and vanilloid. The waveform of ATP-activated inward currents was related to the cell diameter. The I-V curves for both small and medium sized cells manifested obvious inward rectification. Furthermore, we studied the kinetic features of ATP-activated currents and the effects of P2 purinoceptor agonists and antagonists on I(ATP). The findings suggest that ATP receptor-ion channels are expressed differently among different types of rat TG neurons.
Adenosine Triphosphate ; metabolism ; Animals ; Animals, Newborn ; Neurons ; metabolism ; physiology ; Patch-Clamp Techniques ; Rats ; Rats, Sprague-Dawley ; Receptors, Purinergic P2X ; physiology ; Trigeminal Ganglion ; cytology ; metabolism ; physiology

BACKGROUNDThere are few effective methods for treating injuries to the lower trunk of brachial plexus, and the curative effect is usually poor. The purpose of this study was to provide anatomic references for transferring the brachialis muscle branch of musculocutaneous nerve (BMBMCN) for selective neurotization of finger flexion in brachial plexus lower trunk injury, and to evaluate its clinical curative effects.

METHODSMicroanatomy and measurement were done on 50 limbs from 25 adult human cadavers to observe the origin, branch, type of the BMBMCN and median nerve, as well as their adjacent structures. Internal topographic features of the fascicular groups of the median nerve at the level of the BMBMCN were observed. In addition, the technique of BMBMCN transfer for selective neurotization of finger flexion of the median nerve was designed and tested in 6 fresh adult human cadavers. Acetylcholinesterase (AchE) staining of the BMBMCN and median nerve was done to observe the features of the nerve fibers. This technique was clinically tried to restore digital flexion in 6 cases of adult brachial plexus lower trunk injury. These cases were followed up for 3, 6, 9 and 12 months postoperatively. Recovery of function, grip strength, nerve electrophysiology and muscle power of the affected limbs were observed and measured.

RESULTSThe brachialis muscle was totally innervated by the musculocutaneous nerve (MCN). Based on the Hunter's line, the level of the origin of the BMBMCN was (13.18 +/- 2.77) cm. AchE histochemical staining indicated that the BMBMCN were totally made up of medullated nerve fibers. At the level of the BMBMCN, the median nerve consistently collected into three fascicular groups as shown by microanatomy in combination with AchE stain. The posterior fascicular group was mainly composed of anterior interosseous nerves and branches to the palmaris longus. The technique was tested in six fresh cadavers successfully, except that stoma split occurred in one case. Five of the six cases recovered digital flexion 12 months after operation, and at the same time grip strength, muscle power, and nerve electrophysiology also recovered markedly.

CONCLUSIONSThe technique of transferring the BMBMCN for selective neurotization of finger flexion is anatomically safe and effective, with satisfactory clinical outcomes.

Acetylcholinesterase ; analysis ; Adult ; Brachial Plexus ; anatomy & histology ; injuries ; Brachial Plexus Neuropathies ; surgery ; Clinical Trials as Topic ; Female ; Humans ; Male ; Middle Aged ; Musculocutaneous Nerve ; transplantation ; Nerve Transfer ; methods ; Retrospective Studies

OBJECTIVETo discuss the failure reasons of operation for spinal tuberculosis complicated with paraplegia and methods of the second operation.

METHODSSpinal tuberculosis paraplegic patients (18 males, 14 females) were reviewed retrospectively. They have been treated with failing decompressive surgery from January 2001 to December 2006. Seventeen patients received anterior debridement surgery via transpleural approach while the other 15 patients received posterolateral decompression surgery via costotransverse approach. Twenty-two patients got chemotherapy after the surgery.

RESULTSTwenty-three patients were treated by anterior debridement, decompression and graft placement via transpleural approach (9 received the single-stage posterior instrumentation). Five patients received posterolateral debridement and decompression via extrapleural approach. Two patients, recur focus be eliminated. Two patients were given sinus debridement surgery alone. All patients were given anti-tuberculosis chemotherapy. The paraplegia was recovered completely in 26 patients, and partly in 5 patients.

CONCLUSIONSInadequate treatment results in defeated operative. The proper selection of operative modalities and timing on the basis of systematically anti-tuberculosis chemotherapy remains the best mode of therapy for spinal tuberculosis complicated with paraplegia. And it is also essential to choose a radical debridement surgery to decompress the spinal cord and to reconstruct the stability of spine.

Adolescent ; Adult ; Aged ; Child ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Paraplegia ; complications ; Reoperation ; Retrospective Studies ; Treatment Failure ; Tuberculosis, Spinal ; complications ; drug therapy ; surgery

OBJECTIVETo evaluate the association between p53 codon 72 polymorphism (R72P) and the risk of colorectal liver metastases.

METHODSThe p53 R72P genotype was identified by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method in 78 consecutive colorectal cancer patients with liver metastases and 214 age- and sex-matched cases with nonmetastatic colorectal cancer.

RESULTSThe R allele of the p53 R72P polymorphism was more frequently found in metastatic cases than in nonmetastatic cases (P=0.075). Carriers of the 72R allele had a 2.25-fold (95% CI (confidence interval)=1.05 to approximately 4.83) increased risk of liver metastases. On the stratification analysis, 72R-carrying genotype conferred a 3.46-fold (95% CI=1.02 to approximately 11.72) and a 1.05-fold (95% CI=0.36 to approximately 3.08) increased risk of liver metastases for p53 overexpression-positive and negative colorectal cancers, respectively.

CONCLUSIONThese results demonstrate for the first time that the 72R allele of the p53 polymorphism has an increased risk for liver metastases in colorectal cancers positive for p53 overexpression.

Adenocarcinoma ; genetics ; metabolism ; pathology ; secondary ; Case-Control Studies ; Colorectal Neoplasms ; genetics ; metabolism ; pathology ; DNA, Neoplasm ; blood ; genetics ; Female ; Genes, p53 ; Genetic Predisposition to Disease ; Genotype ; Humans ; Liver Neoplasms ; genetics ; metabolism ; secondary ; Logistic Models ; Male ; Middle Aged ; Multivariate Analysis ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics

BACKGROUNDLupus hepatitis is yet to be characterized based on its clinical features and is often difficult to differentially diagnose from other liver diseases. We aimed to elucidate clinical, histopathological and immunopathological features of lupus hepatitis and to evaluate primarily the effectiveness of liver immunopathological manifestations on differential diagnosis of lupus hepatitis from other liver diseases.

METHODSA retrospective study was performed to analyze clinical features of lupus hepatitis in 47 patients out of 504 inpatients with systemic lupus erythematosus (SLE) in First Affiliated Hospital of Sun Yat-sen University, China from May 2006 to July 2009, and to evaluate the association between lupus hepatitis and SLE activity. Additionally, liver histopathological changes by hematoxylin and eosin (HE) staining and immunopathological changes by direct immunofluorescence test in 10 lupus hepatitis cases were analyzed and compared to those in 16 patients with other liver diseases in a prospective study.

RESULTSOf 504 SLE patients, 47 patients (9.3%) were diagnosed to have lupus hepatitis. The prevalence of lupus hepatitis in patients with active SLE was higher than that in those with inactive SLE (11.8% vs. 3.2%, P < 0.05). The incidence of hematological abnormalities in patients with lupus hepatitis was higher than that in those without lupus hepatitis (40.4% vs. 21.7%, P < 0.05), such as leucocytes count (2.92×10(9)/L vs. 5.48×10(9)/L), platelets count (151×10(9)/L vs. 190×10(9)/L), serum C3 and C4 (0.34 g/L vs. 0.53 g/L; 0.06 g/L vs. 0.09 g/L) (P < 0.05); 45 of 47 (95.7%) lupus hepatitis patients showed 1 upper limit of normal (ULN) < serum ALT level < 5 ULN. The liver histopathological features in patients with lupus hepatitis were miscellaneous and non-specific, similar to those in other liver diseases, but liver immunopathological features showed positive intense deposits of complement 1q in 7/10 patients with lupus hepatitis and negative complement 1q deposits in all patients with other liver diseases (Fisher's exact test, P = 0.011).

CONCLUSIONSLupus hepatitis was not infrequent in active SLE patients which would be one of the indices indicating SLE activity. Positive intense deposit of complement 1q in liver may be a characteristic immunopathological feature of lupus hepatitis, which provides a new way to differentially diagnose lupus hepatitis from other liver diseases.

Adolescent ; Adult ; Aged ; Child ; Cohort Studies ; Complement C1q ; analysis ; Female ; Hepatitis, Autoimmune ; etiology ; immunology ; pathology ; Humans ; Liver ; pathology ; Lupus Erythematosus, Systemic ; complications ; Male ; Middle Aged ; Retrospective Studies