According to the sequence of P450 cDNA of Eleutherococcus senticosus, specific primers were designed. Frokaryotic ex pression vector pET30a-P450 was constructed and the prokaryotic expression conditions were optimized. Results showed that the BL21 after being transformed with the recombinant expression vector accumulated the high amount of recombinant protein. SDS-PAGE analysis showed that the recombinant protein was about 53 kDa. The recombinant accumulated the highest amount of recombinant protein af ter IPTG (1 mmol x L(-1)) at 27-37 degrees C for 24 h. Consequently P450 gene of E. senticosus could be expressed successfully by prokaryotic expression vector pET30a-P450. Induction temperature, IPTG concentration, medium type and amount of induction time could all influence the expression of target protein, but the impact strength was different.
Cloning, Molecular ; Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Eleutherococcus ; enzymology ; genetics ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Plant Proteins ; genetics ; metabolism ; Plasmids ; genetics ; metabolism ; Recombinant Proteins ; genetics ; metabolism

OBJECTIVEGas-containing encephalo-meningitis is very rare. There have only been a few cases reported in the literature. The majority of neonatal cases reported in literature died. We report a case of a 5-day-old neonate who developed diffuse pneumocephalus from Enterobacter cloacae septicemia and intracranial infection.

METHODThis neonate was initially treated with penicillin and mezlocillin. He also received phototherapy, intravenous infusion, correction of acidosis and other supportive therapy. Complete blood count, C-Reactive protein, cranial CT scan, blood culture, cerebrospinal fluid culture and biochemistry were tested repeatedly.

RESULTSThis neonate's condition deteriorated after admission. He developed respiratory distress, increased muscle tone and decreased level of consciousness. His WBC and C-reactive protein were elevated, while blood gas, electrolytes, liver enzymes and renal function were within normal range initially. Cranial CT scan was done which demonstrated diffuse pneumocephalus. He was transferred to a higher level hospital for further management at the request of the family. Blood culture done in our hospital subsequently showed growth of Enterobacter cloacae. The infant developed seizures and further deterioration in level of consciousness after transfer. Antibiotics were switched to penicillin and ceftizoxime. Cranial CT scan repeated 2 days after transfer showed hydrocephalus and some resolution of pneumocephalus. Cerebrospinal fluid (CSF) was aspirated from the lateral ventricles two weeks after admission. CSF culture also showed growth of Enterobacter cloacae. Antibiotic was switched to imipenem according to antibiotic sensitivity. His general condition was improved. Blood and CSF cultures were negative 1 month after admission. His head circumference at discharge was 34.6 cm. Repeat cranial CT scan at 4 month of age demonstrated severe hydrocephalus, diffuse leukomalacia and calcification. This infant suffered significant neurodevelopmental deficit. Muscle tone was diffusely increased. Head circumference at 9 month of age was 48.4 cm.

CONCLUSIONThis case suggests the importance of Enterobacter cloacae infection in the newborns. Our analysis of 34 cases of Enterobacter cloacae infection showed that 93.75% - 100% were sensitive to quinolones, 94.12% were sensitive to imipenem, 73.33% were sensitive to gentamicin, 50% were sensitive to piperacillin-tazobactam. Enterobacter cloacae is generally not sensitive to penicillin, first and second generation cephalosporins (0 - 21.4%). Enterobacter cloacae septicemia and intracranial infection in neonates have a high mortality rate and can result in severe neurodevelopmental deficit in survivors.

Enterobacter cloacae ; Enterobacteriaceae Infections ; pathology ; Humans ; Infant, Newborn ; Male ; Meningitis ; microbiology ; Pneumocephalus ; microbiology

OBJECTIVETo investigate the relationship between the extracellular matrix (ECM) and neoplastic progression in hamster with tongue cancer.

METHODSForty-eight specimens of hamster tongue cancer were divided into control group (n=6) and experimental group (n=42). The pathological grade of the specimens was assessed (including 3 stages, namely atypical hyperplasia, carcinoma in situ and early invasive carcinoma). The sections of the tongue were stained with Masson and aldehyde-fuchsin (AF) staining for microscopic observation of the elastic fiber and collagen fiber changes.

RESULTSWithin the connective tissue cores (CTC) of the papillae in the control group was a framework of numerous and fine Gomrori's aldehyde fuchsin-positive elastic fibers. But in the stages of dysplasia and carcinoma in situ, these elastic fibers decreased and further diminished in the CTC in early invasive carcinoma. In dysplasia and carcinoma in situ stages, most of the elastic fibers collapsed with scattered elastic fibers, and the elastic fibers decreased significantly in early invasive carcinoma. The control group showed a significantly greater number of elastic fibers in the experimental group. The collagen fiber was obviously increased and irregularly arranged in dysplasia and carcinoma in situ stage; in early invasive carcinoma, the collagen fibers became thicker with deposition in the lamina propria.

CONCLUSIONAn excessive deposition of collagen fiber and reduction of the elastic fibers is an important factor contributing to the development of tongue carcinoma in hamsters.

Animals ; Carcinoma ; pathology ; Collagen ; metabolism ; Connective Tissue ; pathology ; Cricetinae ; Elastic Tissue ; pathology ; Extracellular Matrix ; pathology ; Neoplasms, Experimental ; pathology ; Tongue Neoplasms ; pathology

OBJECTIVETo investigate the relationship between extracapsular spread (ECS) of cervical metastatic lymph node and the recurrence in patients with oral squamous cell carcinoma (OSCC).

METHODSThe medical records of 74 OSCC patients with histologically confirmed cervical lymph node metastasis were reviewed. They were divided into 2 groups, ECS positive (ECS+) and ECS negative (ECS-). The treatment results were followed up. Statistical analysis, with chi-square test, and multiple logistic regression was conducted.

RESULTSThe overall recurrence rates for pN+/ECS- and pN+/ECS+ patients were 47.6% and 75.0%, respectively, and the cervical recurrence rates for pN+/ECS- and pN+/ECS+ patients were 9.5% and 46.9%, respectively (P < 0.001). Multivariate analysis showed that ECS was one of the independent prognosis factors for cervical recurrence.

CONCLUSIONSExtracapsular spread significantly increased both overall and cervical recurrence rates, and ESC may be a prognosis factor for OSCC patients.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; pathology ; Female ; Follow-Up Studies ; Humans ; Logistic Models ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; pathology ; Male ; Middle Aged ; Mouth Neoplasms ; pathology ; Neck ; pathology ; Neoplasm Recurrence, Local ; Prognosis ; Retrospective Studies

OBJECTIVE: To investigate the relationship between the mutation and abnormal expression of p16 gene in peripheral lung carcinoma and its CT manifestations. METHODS: Immunohistochemistry and PCR-SSCP were used to detect P16 protein expression and p16 gene mutation of 52 cases of peripheral lung cancer. All patients were scanned with spiral CT before the operation and proved by pathology. RESULTS: Of the 52 cases of lung cancer tissues, the negative expression rate of p16 gene protein was 53.8% (28/52), and the deletion or mutation rate of the exon 2 was 23.1% (12/52). There were significant statistical differences of p16 gene defect and its protein loss rates among groups of different clinical stages (P < 0.05), but among groups of different tissue types, different differentiation degree p16 gene defect and its protein loss rates showed no significant statistical difference (P > 0.05). In lung cancer patients with CT appearances of thin spicule, speculated protuberance, pleural indentation, and metastasis of lymph node, p16 gene and its protein loss rates were much higher than those without CT manifestations mentioned above (P < 0.05). However, there were no statistical differences among groups of different tumor sizes, with or without lobulation, with or without cavity, and different contrast enhanced CT values (P > 0.05). CONCLUSION p16 gene mutation and abnormal expression may play an important role in the occurrence and development of lung cancer, and it is relative to CT appearances of lung cancer. p16 gene may be used as a predicting index for clinical diagnosis and prognosis assessment.
Adult ; Aged ; Carcinoma, Squamous Cell ; diagnostic imaging ; genetics ; Female ; Gene Deletion ; Genes, p16 ; Humans ; Lung Neoplasms ; diagnostic imaging ; genetics ; Male ; Middle Aged ; Point Mutation ; Tomography, X-Ray Computed

OBJECTIVESTo summarize the experience of surgical intervention for hepatocellular carcinoma (HCC) with bile duct thrombi (BDT), and to evaluate the influence on prognosis.

METHODSFrom 1994 to 2002, 15 patients with HCC and BDT who underwent surgical intervention were retrospectively analyzed.

RESULTSThe operative procedures included hepatectomy with removal of BDT (n = 7), hepatectomy combined with extrahepatic bile duct resection (n = 4), thrombectomy through choledochotomy (n = 3), and piggy-back orthotopic liver transplantation (n = 1). The 1- and 3-year survival rates were 73.3% and 40%, respectively. Two patients survived over 5 year. The survival rate of patients with portal vein invasion was significantly lower than that of patients without portal vein invasion (P < 0.05).

CONCLUSIONSSurgical intervention was effective for patients with HCC and BDT. Operation after recurrence can prolong the survival time. Liver transplantation is a new operative procedure worthy of study.

Adult ; Aged ; Bile Duct Neoplasms ; surgery ; Carcinoma, Hepatocellular ; complications ; surgery ; Female ; Humans ; Liver Neoplasms ; complications ; surgery ; Liver Transplantation ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Thrombosis ; surgery

BACKGROUNDThis meta-analysis was to determine the association of the cumulative dose of 130/0.4 or 0.42 (hydroxyethyl starch [HES] 130/0.4*) or delta daily fluid balance (i.e., daily fluid balance in HES group over or below control group) with the heterogeneity of risk ratio (RR) for mortality in randomized control trials (RCTs).

METHODSThree databases (PubMed, EMBASE, Cochrane) were searched to identify prospective RCTs reporting mortality in adult patients with sepsis to compare HES130/0.4* with crystalloids or albumin. Meta-analysis was performed using random effects. Sensitivity and meta-regression analyses were used to examine the heterogeneity sources of RR for mortality.

RESULTSA total number of 4408 patients from 11 RCTs were included. The pooled RR showed no significant difference for overall mortality in patients with administration of HES130/0.4* compared with treatment of control fluids (RR: 1.02, 95% confidence interval: 0.90-1.17; P = 0.73). Heterogeneity was moderate across recruited trials (I2 = 34%, P = 0.13). But, a significant variation was demonstrated in subgroup with crystalloids as control fluids (I2 = 42%, P < 0.1). Sensitivity analysis revealed that trials with high risk of bias did not significantly impact the pooled estimates for mortality. Meta-regression analysis also did not determine a dose-effect relationship of HES130/0.4* with mortality (P = 0.298), but suggested daily delta fluid balance being likely associated with mortality in septic patients receiving HES130/130/0.4* (P = 0.079).

CONCLUSIONSInappropriate daily positive fluid balance was likely an important source of heterogeneity in these trials reporting HES130/0.4* associated with excess mortality in septic patients.

Humans ; Hydroxyethyl Starch Derivatives ; therapeutic use ; Randomized Controlled Trials as Topic ; Sepsis ; mortality ; therapy

OBJECTIVETo study the changes of amino acids in cerebral spinal fluid (CSF) in children with spastic or athetotic cerebral palsy (CP) by examining CSF levels of glutamic acid (Glu), gamma-aminobutyric acid (GABA) and aspartate (ASP).

METHODSCSF samples were obtained from 13 children with spastic CP, from 14 children with athetotic CP, and from 10 children without central nervous system and infectious diseases (control group). CSF levels of Glu, GABA and ASP were determined by high-performance liquid chromatography.

RESULTSCSF levels of GABA, ASP and Glu in the control group were 13.04+/-2.19, 10.21+/-0.45 and 8.41+/-2.26 micromol/L, respectively. Compared with the control group, CSF GABA levels in the spastic and the athetotic CP groups (8.02+/-2.03 and 10.01+/-2.68 micromol/L respectively) significantly decreased (P<0.01), whereas CSF levels of Glu (20.99+/-8.15 and 28.77+/-17.62 micromol/L respectively) and Asp (13.53+/-3.93 and 14.02+/-2.88 micromol/L respectively) in the spastic and the athetotic CP groups significantly increased (P<0.01). There were statistical differences in the GABA level between the spastic and the athetotic CP groups (P<0.05). In children with spastic CPCSF Glu level was positively correlated to muscle tension.

CONCLUSIONSCSF excitatory amino acid levels increased, while CSF inhibitory amino acid levels decreased in children with CP. There were differences for CSF amino acid levels in different types of CP. The changes of amino acid levels may contribute to the pathogenesis of CP.

Amino Acids ; cerebrospinal fluid ; Cerebral Palsy ; cerebrospinal fluid ; physiopathology ; Child, Preschool ; Chromatography, High Pressure Liquid ; Female ; Humans ; Male ; Muscle Tonus

OBJECTIVETo study the pattern of lymph node spread in papillary thyroid carcinoma (PTC) with clinically negative node (cN0).

METHODSA total of 106 patients with cN0 PTC who underwent total or subtotal thyroidectomy plus unilateral or bilateral lateral neck dissection (LND, level II-V or level I-V) at West China Hospital of Sichuan University between April 2004 and August 2010 were analyzed retrospectively.

RESULTSThe lateral neck lymph node metastasis in cN0 PTC was significantly associated with sex (male, P = 0.007), tumor stage (T3/T4, P = 0.006), tumor size (> 1 cm, P = 0.014) and the number of positive central lymph nodes (≥ 2, P < 0.001), but not with age and multifocal tumor. Level III (47/116, 40.5%) was the most prevalent metastatic site, followed by level IV (41/116, 35.3%), level II (18/116, 15.5%) and level V (2/29, 6.9%). Of the cases with lymph node metastases in level III and IV, 89.8% (79/88) of primary thyroid tumors existed in the lower and middle sites of the thyroid lobes, while in the cases with lymph node metastases in level II, 77.8% (14/18) of primary thyroid tumors in the upper sites of the thyroid lobes, and 83.3% of cases with level II metastases were accompanied with level III metastases. Two cases with level V metastases were accompanied with metastases in levels II, III and IV.

CONCLUSIONSLND should be considered for cN0 PTC in male, with T3/T4 lesions and positive central lymph nodes ≥ 2, and the range of dissection should include level III and IV. Dissection of level II should be considered in cN0 PTC with primary tumor localized in the upper site of the thyroid lobe or with level III metastasis. Dissection of level V should be considered at present of metastases in level II, III, and IV. For cN0 PTC with tumor size < 1 cm, confined to the thyroid and without lymph node metastasis in the central compartment, LND is not recommended.

Adolescent ; Adult ; Aged ; Carcinoma ; pathology ; Carcinoma, Papillary ; Child ; Female ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; pathology ; Male ; Middle Aged ; Neck Dissection ; Neoplasm Staging ; Retrospective Studies ; Thyroid Neoplasms ; pathology ; Young Adult