Objective The paper analyzes the operating patterns of the outpatient process and identifies the links needing reengineering so as to provide scientific basis for the optimal allocation of outpatient resources and process reengineering and prove the rationality and feasibility of using the queuing theory. Methods Using the methods of the queuing theory, the service time and the patients' arrival time at such service links as registration, billing and accounting, internal medicine, and gynecology in the outpatient department of a certain hospital were measured and the operational indexes at various links such as service intensity, average queuing length, average queuing time, average stay, probability of the service desks being idle, and the probability of the patients having to wait were calculated so as to estimate the rational number of service desks and the optimal value of the sum of the cost of waiting and the cost of service. Results Registration, billing and accounting , service intensity in internal medicine, personnel allocation, and queuing time were basically rational in the outpatient department of the hospital surveyed. One more physician should be added to the gynecology department so as to improve efficiency and reduce patients' waiting time and queuing length. There was currently a shortage of medical staff in the hospital. The cost of hospital input was moderate while the cost of waiting was on the high side. Conclusion It is rational and feasible to evaluate the efficiency of the outpatient process with the methods of the queuing theory. The method deserves to be spread.

OBJECTIVETo establish fluorescence resonance energy transfer (FRET) assay method of detecting proteolytic activity of non-structural protein 3-4A (NS3-4A) serine protease of hepatitis C virus (HCV) for high throughput screening inhibitors against HCV in vitro.

METHODSHCV recombinant plasmid pMAL~c2/NS3-4A was transformed into the E.coli strain K12TB1. Maltose-binding-protein (MBP) NS3-4A fusion protein expression was induced by adding isopropyl-β-D-thiogalacto-pyranoside (IPTG) and purified by affinity chromatography. The proteolytic activity of MBP-NS3-4A protease was analyzed by FRET with the special protease substrate. The reaction system in this model was optimized, and the reliability of the model was evaluated.

RESULTSHigh throughput screening model for HCV NS3-4A protease inhibitors was established, and the best concentrations of enzyme and substrate were optimized. In the model, the Km value of protease was 4.74 μmol/L, Z factor was up to 0.80, and coefficient of variation (CV) was 1.91%. BILN 2061, one of the known HCV protease inhibitors, was measured with the Ki of 0.30 nmol/L.

CONCLUSIONThe assay model using FRET method for HCV NS3 4A serine protease is stable and reliable, and the model is suitable for high throughput screening for HCV NS3 4A protease inhibitors.

Antiviral Agents ; pharmacology ; Drug Evaluation, Preclinical ; Fluorescence Resonance Energy Transfer ; Hepacivirus ; enzymology ; High-Throughput Screening Assays ; methods ; Protease Inhibitors ; pharmacology ; Viral Nonstructural Proteins ; antagonists & inhibitors ; genetics

Severe head injury results in the suppression of cellular immunity associated with dysfunctioning of effector lymphocytes, such as helper T cells(CD4) (and cytotoxic T cells(CD8). Despite progress in the management of increased intracranial pressure following head injury, infection remains the most common complication and the primary cause of prolonged hospitalization and death. This study attempts to assess the cellular immune function following head injury according to the degree of severity, and to establish the clinically available parameters of cell mediated immune(CMI) function, which can then be used for coherent prediction of infection risk. Eighteem head injury patients without severe systemic injury, who divided into three subgroups depending on the severity of head injury, were estimated with the use of CMI multitest kit(Merieux Institute, France) to test delayed-type hypersensitivity(DTH) and enumerated the circulating lymphocyte subpopulation(pan T-cell marker CD3, helper T cell marker CD4, cytotoxic T cell marker CD8 and B-cell marker CD19) on the 1st, 7th, and 21th day of injury. Patients were monitored for evidence of infection for this period. Fourteen patients had no reaction to any antigens of the DTH skin test(anergy) and the remaining four patients had also some degree of anergy. Seven patients became infected and all of them were anergic. There were significant decrease of circulating effector T lymphocytes, both CD4-positive and CD8-positive cells, within 24 hours of injury in the mild as well as the moderate and severe head injury group. CD4-positive cells were nearly completely recovered by the 7th day of injury. CD8-positive cells had sustained significant decrease even after 3 weeks of injury. There was no significant change in pan T-cells(CD3-positive cells) and B-cells(CD19-positive cells). The results suggest that DTH skin test and effector T cell enumeration are both relatively simple and highly sensitive parameters for monitoring CMI function. Especially, anergy of DTH skin test can be used for indicator to predict risk of infection. Mild as well as moderate and severe head injuries may result in the suppression of cellular immunity associated with the dysfunctioning of effector T cell.
B-Lymphocytes ; Craniocerebral Trauma* ; Head* ; Hospitalization ; Humans ; Immunity, Cellular* ; Intracranial Pressure ; Lymphocytes ; Skin ; Skin Tests ; T-Lymphocytes

OBJECTIVETo investigate the effects of the micrometer compound rhizoma coptidis on inflammatory factors and its possible mechanism in rabbit fed with high-lipid food.

METHODThe levels of CRP, IL-1 and TNF-alpha were all determinated by ELISA method. The mRNA and activity of NF-kappaB were determinated by RT-PCR and EMSA, respectively.

RESULTThe level of CRP, IL-1 and TNF-alpha were significantly increased by feeding for 16 weeks with high-lipid diet in rabbit. It was significantly increased that the mRNA and the binging activity with DNA of NF-kappaB in thorax aorta of rabbits fed by high-lipid diet, too. The micrometer compound rhizoma coptidis can reverse the effects of high-lipid diet on CRP, IL-1, TNF-alpha and NF-kappaB.

CONCLUSIONThe micrometer compound rhizoma coptidis can inhibit the expression of inflammatory factor possibly through inhibitting the expression and activity of NF-kappaB.

Animals ; Aorta, Thoracic ; drug effects ; metabolism ; C-Reactive Protein ; metabolism ; Coptis ; chemistry ; Diet, Atherogenic ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Enzyme-Linked Immunosorbent Assay ; Inflammation Mediators ; blood ; Interleukin-1 ; blood ; Male ; Microspheres ; NF-kappa B ; genetics ; metabolism ; Particle Size ; Plants, Medicinal ; chemistry ; Powders ; RNA, Messenger ; genetics ; metabolism ; Rabbits ; Random Allocation ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Necrosis Factor-alpha ; blood

Amyloidosis is the definition for a group of diseases that have, in common, the infiltration of one or more tissues by an abnormal protein material-the amyloid substance, which is detected histologically by their green polarization color after Congo red staining. Despite increased interest on basic nature of amyloidosis by recent immunohistochemical or experimental study, the knowledge about the incidence and neurotoxic effect of cerebral amyloid or concomitant occurrence with brain tumor is still inchoate. We examined the incidence and clinico-pathologic characteristics of the patients with amyloid deposits in supratentorial meningiomas. Particularly about their neurotoxic effect to adjacent brain is considered one of the possible cause of seizure in 33 patients who underwent resection surgery for meningioma at the Keimyung University during the past three years. The pathological review and subgrouping by histologic type were done in all 33 specimens with sufficient size of paraffin block, defined by their morphology and polarization color after Congo red staining for diagnosis of amyloid deposits localized in the tumor. Immunohistochemical studies using monoclonal antibodies for amyloid-A protein(AA) and beta-amyloid(A beta) were evaluated to identify subtypes of amyloidosis. The rate of incidence of amyloid deposit in meningioma was 21%, i.e. seven out of thirty three cases. All laboratory findings and clinical studies did not suggest a systemic form. Seizure occurrence was one out of seven cases(14%), which was of no statistical significance. Immunohistochemical study for AA subtype was all negative, but showed all positive for A beta protein around the vessels. Recent reports has also demonstrated that Amyloid precusor protein(APP) and A beta is related in Alzheimer's disease, hereditary cerebral hemorrhage with amyloidosis-Dutch type(HCHWA-D) and amyloid angiopathy. Our research data indicates that the incidence of amyloid deposit is as high as 21% in supratentorial meningiomas. It seems that it is one of the possible cause of seizure. Nonsystemic microdeposits of amyloid and their subtype and it's relationship to neurotoxic effect in meningiomas remain to be confirmed by immunoelectron microscopic examination or immunohistochemical methods.
Alzheimer Disease ; Amyloid* ; Amyloidosis ; Antibodies, Monoclonal ; Brain ; Brain Neoplasms ; Cerebral Hemorrhage ; Congo Red ; Diagnosis ; Humans ; Incidence ; Meningioma* ; Paraffin ; Plaque, Amyloid* ; Seizures

Purpose: The present study examined the effectiveness of the preoperative ischemia-modified albumin (IMA) levels in predicting the healing period of amputation wounds in patients with diabetes mellitus following a non-traumatic below-knee amputation (BKA). Materials and Methods: This study enrolled 41 diabetic foot ulcer patients who underwent BKA at the authors’ hospital diabetic foot center from April 2016 to April 2022. Among the 41 patients, 29 (70.7%) were male and 12 (29.3%) were female. Their mean age was 64.54±11.38 years (41~81 years). The mean follow-up period was 19.48±5.56 weeks (14~48 weeks) after BKA. The patients were divided into two groups (high IMA group and normal IMA group), which evaluated the healing period, wound dehiscence, and revision operation rate using a Fisher's exact test and Mann–Whitney U test. Three orthopedic surgeons performed stump wound evaluation, and they were evaluated as healing when all sutures were fused without oozing. Results: Thirty patients (73.2%) (group A) showed a high level of IMA (median: 91.2 U/mL), and 11 (26.8%) patients (group B) showed a normal range of IMA (median: 82.7 U/mL). In group A, the median period for wound healing took 1.4 weeks longer, which was significant (p=0.001). No statistical relationship was observed between wound dehiscence, revision operation rate, and IMA value. There was no correlation between the other risk factors (estimated glomerular filtration rate, HbA1c) and the wound healing period. Conclusion Although there was a limitation in using IMA as the sole factor to predict the healing period of amputation wounds in patients after BKA, this study revealed a significant positive correlation between IMA and the period of stump healing after BKA. Therefore, the preoperative IMA levels may help predict the period of stump healing after BKA.

OBJECTIVETo study the effects of different immunodepressants on the sperm parameters of kidney transplant recipients.

METHODSIn 15 healthy fertile men and 37 kidney transplant recipients, ejaculates were aseptically obtained by masturbation. Thirty-seven patients were divided into two groups, 20 patients were treated with Prograf (FK506) combination with mycophenolate mofetil (MMF) and prednisone; 17 patients were treated with cyclosporine (CsA) combination with azathioprine with prednisone. The sperm viability, mobility parameters such as prorsad percentage motility, straight line velocity (VSL), curve line velocity (VCL), velocity of average path (VAP) and morph were estimated with a computer-assisted sperm analyzer (CASA) provided with a multiple-exposure photography system.

RESULTSThere were no significant difference in sperm viability rate [(81.7 +/- 5.7)%, (79.4 +/- 6.8)% and (83.8 +/- 6.0)%], VCL [(24.1 +/- 8.6)%, (23.9 +/- 4.4)%, (24.8 +/- 4.2)% ] and VAP [(19.7 +/- 6.6)%, (18.6 +/- 2.9)%, (21.0 +/- 4.0)%] among groups of FK506, CsA and control, respectively (P > 0.05). The rate of anomaly [(67.8 +/- 5.7)%], the prorsad percentage motility [(46.4 +/- 8.1)%] and VSL [(15.4 +/- 4.6)%] in the group of FK506 were respectively significantly lower and higher than those in the group of CsA [(80.1 +/- 5.6%, (33.3 +/- 6.4)%, (10.2 +/- 2.4)%] (P < 0.05).

CONCLUSIONThe application of FK506 combined with MMF could help recover the mobility and morphology of the sperm in kidney transplantation recipients.

Adolescent ; Adult ; Case-Control Studies ; Cyclosporine ; pharmacology ; Drug Therapy, Combination ; Humans ; Immunosuppressive Agents ; pharmacology ; Kidney Transplantation ; Male ; Mycophenolic Acid ; analogs & derivatives ; pharmacology ; Prednisone ; pharmacology ; Sperm Motility ; drug effects ; Tacrolimus ; pharmacology

BACKGROUNDThe development of regenerative therapies using derivatives of embryonic stem (ES) cells would be facilitated by a non-invasive method to monitor transplanted cells in vivo, for example, magnetic resonance imaging of cells labeled with superparamagnetic iron oxide (SPIO) nanoparticles. Although ES cells have been labeled with SPIO particles, the potential adverse effects of the label have not been fully examined. The objective of this study was to determine whether SPIO labeling affects murine ES cell viability, proliferation, or ability to differentiate into functional endothelial cells (ECs).

METHODSCross-linked iron oxide (CLIO, an SPIO) was conjugated with human immunodeficiency virus transactivator of transcription (HIV-Tat) peptides, and murine ES cells were labeled with either CLIO-Tat, CLIO, or HIV-Tat. After labeling, ES cells were cultured for 4 days and Flk-1(+) ES cells identified and sorted by immunocytochemistry and fluorescence activated cell sorting (FACS). Flk-1(+) cells were replated on fibronectin-coated dishes, and ECs were obtained by culturing these for 4 weeks in endothelial cell growth medium supplemented with vascular endothelial growth factor (VEGF). ES cell viability was determined using trypan blue exclusion, and the proportion of SPIO(+) cells was evaluated using Prussian blue staining and transmission electron microscopy. After differentiation, the behavior and phenotype of ECs were analyzed by reverse transcription-polymerase chain reaction, flow cytometry, immunocytochemistry, DiI-labeled acetylated low-density lipoprotein (AcLDL) uptake, and Matrigel tube formation assay.

RESULTSCLIO-Tat was a highly effective label for ES cells, with > 96% of cells incorporating the particles, and it did not alter the viability of the labeled cells. ECs derived from CLIO-Tat(+) ES cells were very similar to murine aortic ECs in their morphology, expression of endothelial cell markers, ability to form vascular-like channels, and scavenging of AcLDL from the culture medium.

CONCLUSIONSCLIO-Tat is a highly effective label for ES cells and does not adversely affect cell viability, differentiation, or behavior. CLIO-Tat could be a useful marker for the non-invasive monitoring of transplanted stem cells.

Animals ; Cell Differentiation ; drug effects ; Cell Line ; Cell Survival ; drug effects ; Embryonic Stem Cells ; cytology ; drug effects ; Endothelial Cells ; cytology ; drug effects ; Ferric Compounds ; chemistry ; Flow Cytometry ; Immunohistochemistry ; Mice ; Microscopy, Electron, Transmission ; Nanoparticles ; adverse effects ; chemistry ; Reverse Transcriptase Polymerase Chain Reaction

PURPOSE: To investigate the cytotoxic effect of rigid gas permeable(RGP) and reverse geometry lens(RGL) on rabbit corneal cells. METHODS: In 36 white rabbits, right eyes were worn with either RGP or RGL. Left eyes were served as control. The rabbits were sacrificed at 1, 3, 7, 14 days after fitting and 7, 14 days after removal. Morphological changes of the basal epithelial cells and stromal keratocytes were observed using the light and transmission electron microscopy. The samples were examined after TUNEL staining in 3 high power fields (x400). RESULTS: In control eyes, 2.26% of central corneal epithelial cells showed apoptosis. After RGP wearing, the epithelial apoptosis decreased by 0.32% compared to control eyes(2.17%) at 14 days(p<0.001), and returned to 1.55% at 14 days after lens removal(2.04% in control, p=0.161). After RGL wearing, the degree of apoptosis was also lower than control(3.22%) by 0.30% at 14 days(p=0.003). However, it increased to 0.75% at 14 days after lens removal(1.67% in control, p=0.077). A few TUNEL positive fibroblasts were found in superficial stroma in both RGP and RGL group, but not in control eyes. CONCLUSIONS: Both RGP and RGL wearing may decrease the degree of apoptosis in corneal epithelium and lead the apoptosis in corneal stroma. The degree of apoptosis was recovered after lens removal, suggesting the process might to be reversible. The fitting with RGL induced apoptosis of keratocyte more than the fitting with RGP did. It suggests that the RGL wearing may be less physiologic than RGP wearing.
Apoptosis* ; Cornea* ; Corneal Stroma ; Epithelial Cells ; Epithelium, Corneal ; Fibroblasts ; In Situ Nick-End Labeling ; Microscopy, Electron, Transmission ; Rabbits

OBJECTIVETo investigate the pathophysiological role of CXCL16 in immunological liver injury induced by Bacille de Calmette et Guerin (BCG) and lipopolysaccharides (LPS).

METHODSImmunological liver injury was induced by BCG and LPS in mice, and the expression of CXCL16 was detected in the liver tissues by real-time quantitative PCR and immunohistochemical examination. The relationship of the expression of CXCL16 and the extent of hepatic necrosis was investigated histopathologically and immunohistochemically. Mononuclear cells were isolated from the liver tissues and their numbers were counted; T lymphocytes populations in the liver tissue were also analyzed with FACS.

RESULTSThe immunological liver injury model was successfully created. Up-regulation of CXCL16 in injured livers correlated with the extent of liver injury and the amountmononuclear cell infiltrations.

CONCLUSIONThese findings suggest that up-regulation of CXCL16 was closely correlated with liver injury extent during the immunological liver injury induced by BCG-LPS in mice, and intrahepatic recruitment of specific lymphocytes might be an important mechanism of liver injury.

Animals ; Chemical and Drug Induced Liver Injury ; Chemokine CXCL16 ; Chemokine CXCL6 ; Chemokines, CXC ; biosynthesis ; genetics ; Lipopolysaccharides ; Liver Diseases ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Mycobacterium bovis ; Receptors, Scavenger ; biosynthesis ; genetics