Factorial design and response surface techniques were used to design and optimize increasing P450 BM-3 expression in E. coli. Operational conditions for maximum production were determined with twelve parameters under consideration: the concentration of FeCl(3), induction at OD(578) (optical density measured at 578 nm), induction time and inoculum concentration. Initially, Plackett-Burman (PB) design was used to evaluate the process variables relevant in relation to P450 BM-3 production. Four statistically significant parameters for response were selected and utilized in order to optimize the process. With the 416C model of hybrid design, response surfaces were generated, and P450 BM-3 production was improved to 57.90x10(-3) U/ml by the best combinations of the physicochemical parameters at optimum levels of 0.12 mg/L FeCl(3), inoculum concentration of 2.10%, induction at OD(578) equal to 1.07, and with 6.05 h of induction.
Bacillus megaterium ; enzymology ; genetics ; Bacterial Proteins ; biosynthesis ; genetics ; Biotechnology ; Cytochrome P-450 Enzyme System ; biosynthesis ; genetics ; Escherichia coli ; enzymology ; genetics ; Fermentation ; Mixed Function Oxygenases ; biosynthesis ; genetics ; NADPH-Ferrihemoprotein Reductase ; Recombinant Proteins ; biosynthesis ; genetics

The study is to report the establishment of a method of screening the antitumor compounds based on the dynamic bio-response profile of cells to make up for the shortages of conventional end-point tests such as tedious operation and low sensitivity. Based on the principle of electric impedance of cells, the real-time cell electronic sensing (RT-CES) system was used to monitor the effect of epirubicin (EPI), cisplatinum (DDP) and carboplatin (CBP) on the growth of HepG2 cells, with the cell index (CI), half maximal inhibitory concentration (IC50) and detachment curve as evaluation indexes. Meanwhile, cell counting kit-8 (CCK-8) and microscopy were applied for verification. The results showed that CI curve could sensitively real-time profile the inhibitory effect of model drugs on HepG2 cells. The IC50 of EPI, DDP and CBP were 0.53 +/- 0.04, 9.79 +/- 0.26 and 597.00 +/- 3.79 microg x mL(-1), respectively. What's more, the significant differences of detachment curves of the three drugs indicated that their functional mechanisms might be different, this is consistent with the literature. The RT-CES system with non-invasive, label-free and real-time characteristics could be used to monitor the bio-response profile of the three drugs to HepG2 cells, allowing to qualitatively and quantitatively distinguish the antitumor activities of the three drugs, and could be a complementary method for the present screening of antitumor compounds.
Antineoplastic Agents ; pharmacology ; Biosensing Techniques ; methods ; Cell Count ; Cell Line, Tumor ; Cisplatin ; pharmacology ; Drug Screening Assays, Antitumor ; Electric Impedance ; Humans

OBJECTIVETo investigate the incidence of testicular appendages, observe their morphology, and analyze their histopathological origins.

METHODSWe observed 67 testes in 54 patients (15 children and 39 adults) undergoing scrotal surgery, investigated the incidence of testicular appendages, and identified their histopathological origins. We used the Chi-square test to compare the findings from the children and adult patients, with P < 0.05 as statistically significant.

RESULTSThe detection rates of the appendix testis, appendix epididymis, paradidymis, vas aberrans superior, and vas aberrans inferior were 80.6% (54/67), 23.9% (16/67), 1.5% (1/67), 3.0% (2/67), and 1.5% (1/67), respectively. The incidence of testicular appendages was higher in children than in adults (93.3% vs 80.8%), but with no statistically significant difference (Chi2 = 1.339, P > 0.05), and that of the appendix testis and epididymis with pedicles was significantly higher in the former than in the latter (82.4% vs 54.7%, chi2 = 4.149, P < 0.05). Pathological examination showed that the appendix testis originated from the paramesonephric duct, while the appendix epididymis, paradidymis, vas aberrans superior, and vas aberrans inferior from the mesonephric duct.

CONCLUSIONTesticular appendages consist of five embryonic remnants, including appendix testis, appendix epididymis, paradidymis, vas aberrans superior, and vas aber- rans inferior. The appendix testis originates from the paramesonephric duct, and the other four from the mesonephric duct. The clinical implication of these testicular appendages is their tendency to torsion.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Epididymis ; pathology ; Humans ; Infant ; Male ; Middle Aged ; Testis ; pathology ; Young Adult

Objective To assess the accuracy of puncture guided by intelligent positioning (IP) system using magnetic navigation.Methods Five prepared targeted models at three certain depth (<50 mm, samll depth;50-100 mm,medium depth;>100 mm,large depth) underwent puncture guided by intelligent positioning system using IP and conventional ultrasound (US),respectively.Puncture errors,the number of attempt and spent time were recorded and compared .Results For the targets at small,medium and large depth,the errors of IP was (1.88 ±1.18),(1.56 ±0.56) and (3.99 ±1.10) cm,and the errors of conventional US was (4.52 ±2.23),(4.49 ±1.73) and (3.93 ±2.19) cm respectively.The errors of IP were significantly less than those of conventional US at small(t=-2.345,P=0.047) and medium(t=-3.608,P=0.007) depth,but there was no statistically significant difference at large depth (t=0.058,P=0.955). In the IP group,there were statistically significant differences for puncture errors between the small and large depth,as well as between medium and large depth ( F =8.923,P =0.010).There was no statistically significant difference for the errors of IP between the small and medium depth (t=-1.927,P=0.501).For the targets at small,medium and large depth,each puncture was performed in single attempt when guided by IP and in 2,1 and 2 attempt when guided by conventional US .At small and large depth,the numbers of attempt of IP were significantly less than those of conventional US (U=-2.372,P=0.018;U=-2.39, P=0.032).Whereas at medium depth,there was no significant difference (U=-1.000,P=0.690).For the targets at small,medium and large depth,each puncture spent (21.20 ±2.39)s, (27.00 ±4.00)s and (31.80 ±3.83)s when guided by IP,and(45.20 ±9.68),(26.80 ±4.21) and (54.60 ±13.48)s when guided by conventional US.The spent time of IP was less than that with conventional US for small and large depth targets(t =-5.383, P =0.001;t =-3.637, P =0.007).Whereas no statistically significant difference was found for the medium depth target (t=0.077,P=0.916).Conclusion In comparison with conventional US,IP system guided puncture is more accurate and the number of attempt and spent time is less .

OBJECTIVETo observe the alteration of cardiac myocyte nuclear inositol 1,3,4,5-tetrakisphosphate receptor (IP(4)R) binding properties in rat subjected to myocardial ischemic reperfusion in order to further make it clear whether this change is involved in the molecule mechanism of cell apoptosis of rat with myocardial ischemic reperfusion.

METHODSExtracting of cardiac myocyte nucleus was accomplished by saccharose density gradient centrifugation method, the binding properties of nuclear IP(4)R in different conditions were detected by radioligand binding assay. Apoptosis index of myocardial cell was determined by using TUNEL assay.

RESULTS(1) Myocardial cell apoptosis index in rat heart underwent 30 min regional ischemia and 3 h reperfusion increased distinctly compared with that in control group (P < 0.01). (2) There were two IP(4) binding sites located to the nuclear envelope. (3) In ischemic reperfusion injury (IRI) group, Bmax from high affinity binding site of nuclear IP(4)R significantly increased compared with that in sham-operated group, whereas Bmax from low affinity binding site didn't change. Kd values of both sites were all significantly decreased by 63% and 55%, respectively. (4) Phosphorylation of nuclear IP(4)R by PKC increased markedly its binding ability both in IRI and control group (P < 0.05), which was more apparent in IRI group. (5) In sham-operated group, the binding ability of nuclear IP(4)R increased with increasing free calcium concentrations in cytoplasm, and the binding properties of IP(4)R in IRI group were also increased in the condition of calcium overloading.

CONCLUSIONThe increasing of binding properties of nuclear IP(4)R from ischemic reperfusion heart may be one of important mechanism involved in myocardial cell apoptosis, furthermore resulting in myocardial IRI.

Animals ; Apoptosis ; Cell Nucleus ; metabolism ; Male ; Myocardial Reperfusion ; Myocardial Reperfusion Injury ; metabolism ; pathology ; Myocytes, Cardiac ; metabolism ; Rats ; Rats, Wistar ; Receptors, Cytoplasmic and Nuclear ; metabolism

OBJECTIVETo explore the effects of integrative medicine (IM) rehabilitation protocolon motor function, activity of daily living, and quality of life (QOL) in hemiplegia patients after stroke.

METHODSTotally 120 patients with post-stroke hemiplegia were allocated to four groups using sealed envalope drawing, i.e., the rehabilitation group, the Chinese medical treatment group, the acupuncture group, and the comprehensive rehabilitation group, 30 cases in each group. Based on routine rehabilitative training, patients in the Chinese medical treatment group, the acupuncture group, and the compre-hensive rehabilitation group received standardized treatment based on syndrome typing, Shi's Consciousness-Restoring Resuscitation acupuncture, Chinese herbs + acupuncture comprehensive rehabilitatino protocol, respectively. The treatmet cycle consisted of 4 weeks with 24-week follow-ups. Fugl-Meyer motor assessment (FMA), Modified Barthel Index (MBI), and Stroke-Specific Quality of Life Scale(SS-QQL), and safety assessment were taken as main effect indices before treatment, at week 4 of treatment, at week 12 and 24 of follow-ups, respectively.

RESULTSThere was no statistical difference in FMA score, MBI score, SS-QOL score among the four groups before treatment (P > 0.05). These scores were significantly improved in the four groups at week 4 of treatment, week 12 and 24 of follow-ups, respectively (P < 0.05). Besides, FMA score and SS-QOL score were significantly improved in the comprehensive rehabilitation group at each corresponding time point, as compared with other treatment groups (P < 0.05).

CONCLUSIONSThe comprehensive protocol could significantly improve motor function, activity of daily living in hemiplegia patients after stroke, and further improve their QOL. Its effect was better than other single treatment.

Activities of Daily Living ; Acupuncture Therapy ; Hemiplegia ; rehabilitation ; Humans ; Integrative Medicine ; methods ; Medicine, Chinese Traditional ; Motor Skills ; Quality of Life ; Stroke Rehabilitation ; Treatment Outcome

OBJECTIVETo explore the quality of life among people living with HIV/AIDS (PLWHA) and analyze the influencing factors in Kunming.

METHODSA cross-sectional survey was conducted from March to April in year 2011, and 247 PLWHA were selected by convenience sampling from Yunnan CDC, Yunnan and Kunming infectious disease hospitals. General questionnaires, the simplified Chinese edition of medical outcomes study-HIV health survey (MOS-HIV, including 11 dimensions) and Social Support Scale were used. t test and multivariable linear regression model were used to analyze the results.

RESULTSFor the subjects investigated, the age was (39.8 +/- 11.9) years old, the median (quartile) value of symptoms related to HIV infection was 1(2). As to the scores of quality of life measured by MOS-HIV, physical summary score was 47.4 +/- 11.2, mental summary score was 43.6 +/- 9.7; for the scores of 11 dimensions of the MOS-HIV,that were general health (42.9 +/- 19.9), physical function (79.4 +/- 24.9), role function (59.8 +/- 48.2), social function (67.0 +/- 33.6), cognitive function (71.0 +/- 25.4), pain (81.3 +/- 26.2), mental health (62.0 +/- 22.3), vitality (49.3 +/- 23.8), health distress (74.4 +/- 21.0), quality of life (51.8 +/- 21.1), health transition (49.0 +/- 29.8). The total score of social support was 28.6 +/- 7.6, of which the score of subjective social support was 17.2 +/- 6.3, the score of the objective social support was 5.9 +/- 2.2; the score of the utilization of social support was 5.5 +/- 1.9. Multivariable linear regression analysis showed that the more the symptoms related to HIV infection, the lower the physical summary scores (standardized coefficients b' = -0.22), the general health (b' = - 0.31), the physical function (b' = -0.16), the role function (b' = -0.23), the pain (b' = -0.21), the mental health (b' = -0.22), the vitality (b' = -0.22) and the health distress scores (b' = - 0.24) (all P values < 0.05); the older the age, the lower the physical summary scores (b' = - 0.16), the mental summary scores (b' = - 0.16), the physical function (b' = -0.26), the vitality (b' = -0.26) and the quality of life scores (b' = -0.17) (all P values < 0.05); the higher the score of the subjective social support,the higher the physical summary scores (b' = 0.26), the mental summary scores (b' = 0.22), the general health (b' = 0.27), the social function (b' = 0.26), the mental health (b' = 0.15) and the quality of life scores (b' = 0.22) (all P values < 0.05); the higher the score of the utilization of social support, the higher the physical function (b' = 0.16) and the health transition scores (b' = 0.31) (all P values < 0.05).

CONCLUSIONPLWHA in Kunming have relatively lower scores of quality of life. A large number of symptoms during infection, older age and lower score of subjective social support were the hazard factors of quality of life in PLWHA.

Acquired Immunodeficiency Syndrome ; epidemiology ; Adult ; China ; epidemiology ; Cross-Sectional Studies ; Factor Analysis, Statistical ; Female ; HIV Infections ; epidemiology ; Humans ; Male ; Middle Aged ; Quality of Life ; Social Support ; Surveys and Questionnaires

OBJECTIVETo investigate oxidative DNA damage in pharmacy technicians preparing antineoplastic drugs at the PIVAS (Pharmacy Intravenous Admixture Service) in two Chinese hospitals.

METHODSUrinary 8-OHdG served as a biomarker. 5-Fluorouracil (5-FU) concentrations in air, masks and gloves were determined. The spill exposure of each PIVAS technician to antineoplastic drugs was investigated. Eighty subjects were divided into exposed group I, II, and control group I, II.

RESULTS5-FU concentration ratios for gloves and masks in exposed group I were significantly higher than those in exposed group II (P<0.05 or P<0.01). The average urinary 8-OHdG concentrations in exposed group I, control group I, exposed group II, and control group II were 14.69±0.93, 10.68±1.07, 10.57±0.55, and 11.96±0.73 ng/mg Cr, respectively. Urinary 8-OHdG concentration in exposed group I was significantly higher than that in control group I or that in exposed group II (P<0.01). There was a significant correlation between urinary 8-OHdG concentrations and spill frequencies per technician (P<0.01).

CONCLUSIONThere was detectable oxidative DNA damage in PIVAS technicians exposed to antineoplastic drugs. This oxidative DNA damage may be associated with their spill exposure experience and contamination of their personal protective equipment.

Adult ; Air ; analysis ; Antineoplastic Agents ; analysis ; toxicity ; Case-Control Studies ; China ; DNA Damage ; Deoxyguanosine ; analogs & derivatives ; urine ; Female ; Fluorouracil ; analysis ; toxicity ; Gloves, Protective ; Health Personnel ; statistics & numerical data ; Hospitals ; statistics & numerical data ; Humans ; Male ; Masks ; Occupational Exposure ; statistics & numerical data ; Oxidative Stress ; Young Adult

OBJECTIVETo study the morphologic and immunohistochemical features of gastrointestinal B-cell lymphomas.

METHODSOne hundred and ninety-four cases of gastrointestinal B-cell lymphoma were retrieved from the archival file. The clinical features and pathologic findings were reviewed. Immunohistochemical study for B-cell markers, T-cell markers, bcl-6, CD10, bcl-10, cyclin D1, TdT, MUM1 and Ki-67 was carried out.

RESULTSThe male-to-female ratio was 1.4:1. The age of patients ranged from 8 to 85 years. Amongst the 194 cases studied, 128 (66.0%) were diagnosed as diffuse large B-cell lymphoma, including 16 cases of large cell lymphoma associated with mucosa-associated lymphoid tissue (MALT) lymphoma component. There were also 40 cases (20.6%) of MALT lymphoma, 8 cases (4.1%) of follicular lymphoma, 5 cases of (2.6%) of lymphoplasmacytic lymphoma, 3 cases (1.6%) of mantle cell lymphoma, 1 case of (0.5%) of B-lymphoblastic lymphoma and 9 cases (4.6%) of indefinite type (including 5 biopsy cases). The site of involvement included stomach (100 cases, 51.5%), small intestine (43 cases, 22.2%), ileocecal junction (26 cases, 13.4%), appendix (1 case, 0.5%), colon (21 cases, 10.8%) and rectum (3 cases, 1.6%). Amongst the 163 cases which had undergone surgical resection, 20 cases (12.3%) cases had invasion down to the mucosa, 20 cases (12.3%) down to the superficial muscular layer, 19 cases (11.6%) down to the deep muscular layer and 104 cases (63.8%) with full-thickness involvement. Histologic examination showed lymphoepithelial lesions in 52 cases, residual lymphoid follicles in 29 cases, coagulative necrosis in 66 cases and nodular growth pattern in 30 cases. The lymphoma cells in all cases were immunoreactive for B-cell marker CD20. There was also various degrees of positivity for bcl-6, CD10, bcl-10, cyclin D1, TdT, MUM1 and Ki-67.

CONCLUSIONSGastrointestinal B-cell lymphomas can be subdivided into two main groups: large B-cell lymphomas and small B-cell lymphomas. The latter group often poses diagnostic pitfalls. Accurate pathologic typing requires correlation with histologic and immunohistochemical findings.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD20 ; metabolism ; Child ; DNA-Binding Proteins ; metabolism ; Female ; Gastrointestinal Neoplasms ; metabolism ; pathology ; surgery ; Humans ; Immunohistochemistry ; Lymphoma, B-Cell ; metabolism ; pathology ; surgery ; Lymphoma, B-Cell, Marginal Zone ; metabolism ; pathology ; surgery ; Lymphoma, Follicular ; metabolism ; pathology ; surgery ; Lymphoma, Large B-Cell, Diffuse ; metabolism ; pathology ; surgery ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neprilysin ; metabolism ; Proto-Oncogene Proteins c-bcl-6 ; Young Adult

OBJECTIVETo summarize our 10-year experience with percutaneous fluoroscopically guided removal (PFGR) of metallic foreign body (MFB) in the soft tissue.

METHODSPFGR was performed in 65 patients for removing a total of 368 MFBs from the soft tissues. The MFBs ranged from 0.2 to 0.3 cm in length embedded in the soft tissue for 7 days to 8 years. For superficial MFBs, the MFBs were removed directly with curved forceps under real time fluoroscopy. For deep MFBs, trocar technique was applied using the instruments for percutaneous diskectomy, with the outer cannula inserted toward the foreign body under real-time fluoroscopy followed by MFB removal by grasping forceps.

RESULTSA total of 346 MFBs were successfully removed without any serious complications (success rate 94.0%), including 154 removed directly and 192 MFBs with trocal technique. The time of the procedures and radiation exposure for MFB removal was 30 s to 20 min and 1-6 min, respectively.

CONCLUSIONPGFR of MFBs in the soft tissue under fluoroscope is safe and effective. Direct removal using curved forceps is suitable for MFBs in superficial soft tissues, while trocal techniques needs to be utilized for deep MFBs.

Adolescent ; Adult ; Child ; Female ; Fluoroscopy ; methods ; Foreign Bodies ; diagnostic imaging ; surgery ; Humans ; Lower Extremity ; Male ; Metals ; Radiography, Interventional ; Young Adult