Calcinosis ; pathology ; surgery ; Child ; Diagnosis, Differential ; Humans ; Knee ; Male ; Skin Diseases ; pathology ; surgery

ObjectiveTo investigate the relationship between the degenerative mechanisms of lumbar intervertebral disc (LID) and apoptosis.MethodsThe total RNAs were isolated from human LID tissues. Both the mRNAs from the degeneration and normal LID were reversely transcribed to the cDNAs. The cDNAs were labeled with the incorporations of fluorescent dUTP, for preparing the hybridization probes. The mixed probes were then hybridized to the cDNA microarray. After high-stringent washing, the cDNA microarray was scanned for the fluorescent signals and analyzed by computer image analysis. The apoptotic status and the expression of Bcl-2 and Bax in 12 cases of degenerative LID and 10 cases of normal LID were detected with TdT-mediated dUTP-biotin nick end labeling (TUNEL) and immunohistochemistry methods.ResultsAmong the 4096 targets, there were 10 genes related to apoptosis. The expression related to Bax protein gene was up-regulated and it was down-regulated for Bcl-2 protein. In group of normal LID, the average apoptotic index (AI) was (24.897±3.620); percentage of Bcl-2 positive cells was (31.440±4.150)%; percentage of Bax positive cells was (29.372±2.588)%, average optical density (OD) values of positive particles were (0.183± 0.010 ), ( 0.203 ±0.012) and (0.169±0.005) respectively. In group of degenerative LID, the average AI was (49.232±3.440); percentage of Bcl-2 positive cells was (18.239±2.470)%; percentage of Bax positive cells was (52.349±3.764)%; average OD values of positive particles were (0.152±0.003), (0.310±0.008) and (0.262±0.014) respectively. There were significantly differences in AI and expressions of Bcl-2 and Bax proteins between normal LID and degenerative LID (P<0.05).ConclusionCell apoptosis plays an important role in the process of LID degeneration. Both Bcl-2 and Bax take part in the occurrence and progression of LID.

OBJECTIVETo evaluate the correlation between the mean of time blood flow velocity (TVmean) of feeding artery around hepatocellular carcinoma (HCC) and the microvessel density (MVD) in relation to tumor cell differentiation, and to evaluate the usefulness of TVmean as a noninvasive preoperative marker of biologic characteristic of HCC.

METHODSTo measure TVmean of feeding arteries around the HCC in 45 patients before operation and TVmean of minute arteries in hepatic tissue in 45 normal subjects by ultrasonographic examination. Tumor cell differentiation grade was determined by routine histopathological staining. Tumor MVD was measured by immunohistochemical staining with monoclonal antibodies against F VIII RAg of excised HCC. Correlationship between TVmean of feeding arteries and MVD of HCC was studied.

RESULTSA linear correlation between TVmean of feeding arteries around the tumors of HCC and MVD of HCC tissue (r = 0.794, y = 18.2764 + 1.5544x) was obtained. With TVmean > or = 21.3 cm/s, the positive and negative predictive values of poorly differentiated HCC were 97.4% and 87.5%, respectively. With TVmean < 21.3 cm/s, those of well differentiated HCC were 88.5% and 97.3%, respectively. The degree of reliability of calculating HCC cell differentiation according to TVmean was 89.5%.

CONCLUSIONTumor growth and cellular differentiation, the two major factors affecting prognosis of HCC, may be evaluated by TVmean of feeding arteries around the tumor. It offers a useful preoperative information for predicting prognosis of HCC patients.

Adult ; Aged ; Arterioles ; pathology ; Blood Flow Velocity ; Capillaries ; pathology ; Carcinoma, Hepatocellular ; blood supply ; pathology ; physiopathology ; Female ; Humans ; Liver Neoplasms ; blood supply ; pathology ; physiopathology ; Male ; Middle Aged ; Neovascularization, Pathologic ; pathology

OBJECTIVETo evaluate the effect of a 10-day course of triptolide (TP) on rat cytochrome (CY) P3A4 activity, and on the pharmacokinetics of cyclophosphamide (CPA).

METHODSIn the pharmacokinetics experiment, rats were orally given 0.9% NaCl solution (n=5) and TP [1.2 (mg/kg·d)] for 10 days and a single dose of CPA was administered intravenously (100 mg/kg) to rats on day 11. Blood samples were collected up to 4 h at predetermined time intervals, the plasma concentration of CPA was determined by high performance liquid chromatography (HPLC) and pharmacokinetic parameters were determined. In the in vitro CYP3A4 activity inhibition research, rat blank liver microsomes were divided into 3 groups: a control group, a TS (5 μL, 200 μmol/L) with TP (5 μL, 12.5 μmol/L) group, a TS with ketoconazole (5 μL, 1 μmol/L) group. Concentration of 6β-hydroxylated testosterone (6β-OHT) in liver microsomes was measured by HPLC and the activity of CYP 3A4 was calculated through the following formula: Einhibitor/Econtrol × 100%=Cinhibitor/Ccontrol × 100%.

RESULTSCompared with the control group, the area under the plasma concentration-time curve (AUC0-∞) of CPA was significantly increased by 229.05% pretreated with TP (P<0.01). Peak plasma concentrations (Cmax) of CPA was significantly increased and plasma half-life was correspondingly extended. The CYP3A4 activity was significantly inhibited by ketoconazole 93.5%±0.2% and TP 84.6%±0.3% compared with the control group (P<0.01 and P<0.05, respectively).

CONCLUSIONOur results strongly suggested that long-term oral intake of TP can distinctly inhibit the CYP3A4 activity and this inhibition evidently decrease the formation of toxic metabolites of CPA.

Animals ; Cyclophosphamide ; pharmacokinetics ; Cytochrome P-450 CYP3A ; metabolism ; Cytochrome P-450 CYP3A Inhibitors ; pharmacology ; Diterpenes ; pharmacology ; Epoxy Compounds ; pharmacology ; Herb-Drug Interactions ; Hydroxytestosterones ; metabolism ; Immunosuppressive Agents ; pharmacokinetics ; Injections, Intravenous ; Ketoconazole ; pharmacology ; Male ; Microsomes, Liver ; drug effects ; metabolism ; Phenanthrenes ; pharmacology ; Rats, Sprague-Dawley

OBJECTIVETo study the effects of gonadotroph-releasing hormone (GnRH) agonist (GnRH-a) and GnRH antagonist (GnRH-ant) on cyclophosphamide (CTX)-induced follicle apoptosis in female rats.

METHODSThirty-six female Sprague- Dawley rats were randomized into 6 groups, namely normal saline (NS), CTX, GnRH-a+NS, GnRH-a+CTX, GnRH-ant+NS, and GnRH-ant+CTX groups. The rats were sacrificed between the first and second week after the treatments., and the follicle apoptosis was investigated using TUNEL assay and transmission electron microscopy.

RESULTSThe apoptosis rate of the granulose cells in the follicles in late development was significantly higher than that in early follicles, and the apoptosis rate of the oocytes and granulose cells in rats with CTX treatment was significantly higher than that in rats without CTX treatment (P<0.05). The apoptosis rate of the granulose cells in GnRH-a groups (ranging from 33.40 - or + 4.59 to 73.25 - or + 5.35) was significantly higher than that in GnRH-ant groups (27.46 - or + 4.52 to 49.38 - or + 5.02, P<0.05), but there was no significant difference in the oocytes of early follicles between GnRH-a groups (23.48 - or + 4.25 to 36.15 - or + 4.23) and GnRH-ant groups (21.47 - or + 3.81 to 34.04 - or + 5.54, P>0.05). Electron microscopy revealed characteristic apoptotic changes of the oocytes in early follicles and granulose cells in early and late follicles. The apoptotic changes were especially typical in the granulose cells showing the formation of the apoptotic bodies, and the oocytes only showed chromatin condensation and aggregation.

CONCLUSIONIn the rat mode, GnRH-a promotes while GnRH-ant suppressed follicle apoptosis induced by CTX. GnRH analogues regulates mainly granulose cell apoptosis, but have little effect on oocyte apoptosis.

Animals ; Apoptosis ; drug effects ; Cyclophosphamide ; toxicity ; Female ; Gonadotropin-Releasing Hormone ; analogs & derivatives ; antagonists & inhibitors ; Granulosa Cells ; pathology ; Oocytes ; pathology ; Ovarian Follicle ; pathology ; Random Allocation ; Rats ; Rats, Sprague-Dawley

Objective: To compare the clinical effects of autologous semitendinosus tendon and allogenic tendon arthroscopic anatomical reconstruction of anterior talofibular ligament (ATFL) combined with calcaneofibular ligament (CFL) in the treatment of chronic lateral ankle instability. Methods: A retrospective analysis was made of 55 patients with chronic lateral ankle instability who underwent arthroscopic reconstruction of ATFL combined with CFL from January 2012 to June 2017. A total of 28 cases were treated with autologous semitendinosus tendon (autologous group), including 19 males and 9 females, with an average age of 28.5±8.03 years (range, 16-46 years). A total of 27 cases were treated with allogenic tendon (allogenic group), including 17 males and 10 females, with an average age of 27.48±7.89 years (range, 16-46 years). ATFL/CFL was reconstructed by the same method in both groups. The reconstruction methods were the same between the groups. The talus and calcaneus were fixed with absorbable compression nails. Results: The operation duration in the autologous group was 94.07±7.83 min, which was longer than that in the allogeneic group 63.56±7.96 min (t=14.51, P<0.001). Fever days 5.26±0.90 days in allogeneic group were longer than 2.46±0.74 days in autologous group (t=-12.55, P<0.001). Wound healing duration in allogeneic group was 13.44±3.33 days longer than that in autologous group 10.32±2.34 days (t=-4.01, P<0.001). In the autologous group, 28 cases were followed up for 34.54±16.04 months, and 27 cases in the allograft group were followed up for 42.74±17.79 months. The mean AOFAS score improved from 63.64±11.20 before operation to 90.21±4.48 after operation in the autologous group, and that improved from 63.93±10.59 before operation to 89.56±5.15 after operation in the allogeneic group with no significant difference between the two groups after operation (t=0.506, P=0.615). The mean VAS score decreased from 5.79±1.79 before operation to 1.54±1.35 after operation in the autologous group, and from 5.89±1.78 before operation to 2.04±1.32 after operation in the allogeneic group. There was no significant difference between the two groups after operation (t=-1.396, P=0.168). Tegner score increased from 4.07±1.39 to 6.43±1.14 in the autologous group and from 3.85±1.06 to 6.52±0.85 in the allogeneic group with no significant difference between the two groups after operation (t=-0.333, P=0.740). Stress radiographic showed that the talar tilt angle decreased from 15.60°±3.86° to 6.01°±2.64° in the autologous group, 16.99°±3.78° to 7.14°±3.34° in the allogeneic group, and there was no significant difference between the two groups after operation (t=-1.382, P=0.171). Anterior talar displacement reduced from 10.82±3.12 mm to 4.03±1.69 mm in the autologous group, from 10.10±2.02 mm to 4.17±1.52 mm in the allogeneic group, and there was no significant difference between the two groups after operation (t=-0.326, P=0.746). No donor tendon dysfunction was found in the autologous group. At the end of follow-up, there was no difference in ankle dorsiflexion, plantar flexion and hind foot mobility between autologous group and allogeneic group. Conclusion Arthroscopic autologous tendon and allogeneic tendon reconstruction of AFTL combined with CFL can obtain satisfactory short-term results. The autologous tendon group was superior to the allogeneic group in terms of fever, wound healing time. However, there was no significant difference in clinical effects between the two groups.

In spite of receiving chemotherapy, the response of patients with cancer can be extremely variable. Chemosensitivity testing is being applied in institutes and some hospitals to improve the effects of chemotherapy. It would be useful for choosing the most effective drug and strategy for individual chemotherapy and to exclude the resistance of the tumor cells. In this way, the individualized chemotherapy can be established. Up to today, there are more than 10 approaches established for chemosensitivity testing assays, such as single cell culture assay (including MTT, MTS, ATP), nude mouse model sensitivity examination, collagen gel droplet embedded culture drug sensitivity test and histoculture drug response assay etc. This paper reviews some current methods, and their possibility for directing clinical chemotherapy.
Animals ; Antineoplastic Agents ; therapeutic use ; Cell Culture Techniques ; methods ; Collagen ; Culture Techniques ; methods ; Disease Models, Animal ; Drug Resistance, Neoplasm ; Drug Screening Assays, Antitumor ; methods ; Gels ; Humans ; Mice ; Neoplasm Transplantation ; Neoplasms ; drug therapy ; Precision Medicine ; Sensitivity and Specificity ; Tumor Cells, Cultured

Objective To investigate the effects of selective cyclooxygenase 2 inhibitor on the hyperplasia of parathyroid glands from uremic rats. Methods Sixty-five 5/6-nephrectomized (Nx) and fifteen sham operated rats were assigned to 4 groups: (1)Sham group (n=14):shamoperated +normal phosphate diet (P 0.8％,Ca 1.2％)； (2) Nx-HP group (n=17):Nx+high phosphate(HP) diet (P 1.2％,Ca 1.2％)； (3)Prophylactic COX2 inhibition group (Prey group,n=18):Nx+HP+celecoxib 100 mg· kg-1·d-1 for 3 months； (4)Therapeutic group (Ther group,n=18):Nx+HP+celecoxib 100 mg·kg-1·d-1 starting at the second month of the 5/6 nephrectomy.At the end of 3 month,blood,urine and parathyroid samples were collected.The expressions of COX2 and PCNA were determined by immunohistochemistry,Western blotting and real-time PCR. Results All of the Nx rats fed with high phosphate diet for 3 months manifested progressively increasing serum creatinine,serum iPTH as well as augmentation of parathyroid gland volume,suggesting that secondary parathyroid hyperplasia animal model was established successfully.Celecoxib significantly decreased serum iPTH levels [Sham (34.77±0.83),Nx-HP(100.73±4.35),Prey (87.36±2.18),Ther (87.47±1.76) ng/L,P＜0.05],the size of the parathyroid glands in Nx rats [Sham (0.461±0.089),Nx-HP (2.436±0.372),Prey (0.987±0.254),Ther (1.27±0.305) mm2/kg,P＜0.05] and PCNA expression in PG determined by Western blotting (decreased to 52.91％ in Prev group and 34.68％ in Ther group respectively,P＜0.05).No significant difference was observed between the two COX2 inhibition groups.The levels of COX2 expression in parathyroid gland were greatly increased in three Nx groups compared with that in sham group (2.47-fold in Nx-HP,2.34-fold in Prey group,3.04-fold in Ther group,P＜0.05).COX2 inhibitor had no effects on COX2 expression in PGs.Real-time PCR analysis demonstrated the same trends of mRNA expression of COX2 and PCNA in PGs of rats. Conclusion Selective inhibition of COX2 may help to suppress the hyperplasia of parathyroid glands in uremic rats.

OBJECTIVETo improve the differential diagnosis of tuberculous meningitis (TBM) and reduced potential misdiagnosis of TBM.

METHODSThe clinical data of 47 misdiagnosed cases of TBM between January, 1994 and June, 2009 were investigated retrospectively. The clinical presentations and causes for the misdiagnoses were analyzed.

RESULTSThe 47 patients with misdiagnosed TBM included 28 male and 19 female patients with a mean age of 36.84∓16.41 years. Eight patients had an acute onset, 10 had a subacute onset, and 29 had chronic disease. The initial symptoms, in the descending order of their frequencies, included fever and headache (87.2%), anergia and dyskinesia (27.7%), cerebral nerve damage (23.4%), decreased level of consciousness (14.9%), and urinary and fecal incontinence (2%). Meningeal irritation was present in 25 cases and positive Babinksi sign was found in 19 cases. Elevated intracranial pressure occurred in 51.1% of the cases, and 16 cases showed papilloedema. Non-purulent CSF with elevated protein was found in 86.7%, decreased glucose in 50%, and decreased chlorinate in 53.3% of the cases. Eight out of 23 cases showed a positive result of PPD test. MRI identified abnormitis with meningeal enhancement in 15 cases, hydrocephalus in 7 cases and infarction in 14 cases. Tuberculoma was found in 2 cases, and spinal cord lesions were found 4 cases. All the patients received anti-tuberculosis therapy, which resulted in symptomatic improvement in 39 cases, fluctuated condition in 2 cases; 5 patients discontinued the treatment and 1 died.

CONCLUSIONEarly TBM often presents with atypical features and its differential diagnosis can be difficult. CSF monitoring and careful inspection of the radiographic data can be helpful in the diagnosis of suspected cases, for which early anti-TB treatment is an important means to reduce misdiagnosis.

Adult ; Diagnosis, Differential ; Diagnostic Errors ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Tuberculosis, Meningeal ; cerebrospinal fluid ; diagnosis ; Young Adult

OBJECTIVETo investigate the nucleotide-binding oligomerization domain-2 (NOD-2) gene expression in deep caries and the effects of NOD-2 agonist muramyl dipeptide (MDP) on the differentiation of human dental pulp cells (hDPC).

METHODSNOD-2 gene level in deep caries and healthy pulp tissue was determined by real-time quantitative polymerase chain reaction (realtime-PCR). Realtime-PCR, Western blotting and immunofluorescence were performed to evaluate NOD-2 gene and protein expression. Dentin sialoprotein (DSP) protein level was assessed when hDPC were challenged by different concentrations of MDP for 24 hours, and sialophosphoprotein (DSPP), osteocalcin (OCN) mRNA and osteopontin (OPN) protein level were detected at different time points after incubation with 0.1 mg/L MDP.

RESULTSNOD-2 mRNA level was higher in pulp tissue of deep caries (0.2610 ± 0.0824) than that in healthy controls (0.0024 ± 0.0002), P < 0.05. The expression of NOD-2 gene and protein increased in a time denpendent manner upon stimulation with MDP. Immunofluorescence confirmed that NOD-2 protein was located in cytoplasm. Moreover, 0.1 mg/L MDP augmented DSP protein level. DSPP and OCN mRNA were elevated with time and reached the peak at 12 h and down-regulated. OPN protein level also increased with time.

CONCLUSIONSDental pulp NOD-2 expression are up-regulated in pulp tissue of deep caries. MDP may be related to the differentiation of hDPC.

Acetylmuramyl-Alanyl-Isoglutamine ; pharmacology ; Adjuvants, Immunologic ; pharmacology ; Adolescent ; Adult ; Cell Differentiation ; drug effects ; Cells, Cultured ; Dental Caries ; pathology ; Dental Pulp ; cytology ; metabolism ; Extracellular Matrix Proteins ; genetics ; metabolism ; Gene Expression ; Humans ; Nod2 Signaling Adaptor Protein ; genetics ; metabolism ; Osteocalcin ; genetics ; metabolism ; Osteopontin ; genetics ; metabolism ; Phosphoproteins ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Sialoglycoproteins ; genetics ; metabolism ; Young Adult