AIM: To investigate the effect of curcumin on the binding activity of hepatic nuclear factor-kappa B(NF-?B) and the expression of peroxisome proliferator-activated receptor-?(PPAR-?) in rats with liver fibrosis induced by carbon tetrachoride. METHODS: A total of 60 clean male rats were randomly and averagely divided into group A,B,C,D,E and F.The rats in group A served as normal controls,while those in other five groups were injected subcutaneously 40% CCI_4 for seven weeks to induce the model of liver fibrosis.After seven weeks,the rats in group C,D,E,F were intragastrically administered with 50 mg/kg silibinin,100 mg/kg cur,200 mg/kg cur,400 mg/kg cur once per day for six weeks,respectively.HE staining was used to observe the pathological changes of liver tissues under light microscope,and immunohistochemistry and reverse transcriptase polymerase chain reaction(RT-PCR) were performed to detect the activity of NF-?B,PPAR-? and mRNA expression of PPAR-?.RESULTS: The inflammatory and fibrotic degrees were obviously alleviated in group C,D,and E compared with group B.The expression of NF-?B p65 was significantly decreased in liver tissue in group C,D and E,compared with model control group B(P

Objective:To study the effect of Qingyi Lidan Granule on the serum levels of high mobility group box 1 (HMGB 1),heat shock protein 70 (HSP70),heat shock protein 27 (HSP27) and interleukin-8 (IL-8) of patients with severe acute pancreatitis.Methods:From August 2015 to July 2016,84 patients with severe acute pancreatitis in our hospital were selected and randomly divided into the observation group and the control group according to random number,42 cases in each group.The control group was given routine treatment,and the observation group was treated by Qingyi Lidan Granule on the basis of control group.The recovery of blood amylase to normal time,white blood cell recovery to normal time,recovery of gastrointestinal function to normal time and relieving time of abdominal pain,serum levels of HMGB1,HSP70,HSP72 and IL-8 in both groups were observed and compared before and after treatment.Results:After treatment,the total clinical effective rate of observation group was significantly higher than that of the control group[92.86％ (39/42) vs 71.43％ (30/42)] (P＜0.05).The recovery of blood amylase to normal time,white blood cell recovery to normal time,recovery of gastrointestinal function to normal time and relieving time of abdominal pain in the observation group were significantly shorter than those of the control group (P ＜0.05).Before treatment,no significant difference was found in the serum levels of HMGB 1,HSP70,HSP72,IL-8 between groups (P＞0.05).After treatment,the serum levels ofHMGB1,HSP70,HSP72 and IL-8 in both groups were significantly lower than those before treatment (P＜0.05).The levels ofHMGB1,HSP70,HSP72 and IL-8 in the observation group were lower than those in the control group (P＜0.05).Conclusion:Qingyi Lidan Granule could effectively reduce the levels of serum HMGB 1,H SP70,HSP27 and IL-8 and enhance the clinical curative effect of patients with severe acute pancreatitis.

BACKGROUND: Traumatic osteoarthritis (OA) resulted from the injury of joints and postoperation of joints is commonly observed. Intra-articular injection of sodium hyaluronate (Na-HA) has been considered as effective method for OA. OBJECTIVE: To observe the influence of intra-articular injection of NaHA on mRNA expression of inducible nitric oxide synthase (iNOS) in cartilage of traumatic OA induced by transection of anterior cruciate ligament. DESIGN: Randomized controlled animal experiment. SETTING: Department of Orthopaedics, Renmin Hospital, Wuhan Uni ersity. MATERIALS: The experiment was performed in Laboratory of Depart ment of Orthopaedics, Renmin Hospital, Wuhan University from April to December 2003, in which, 16 clean healthy flat-eared white rabbits, aged 5-6 months were employed. The rabbits were randomly divided into Na- HA injection group and saline control group with 8 rabbits in each group. Na-HA (2000, No 366095) was provided by Shanghai Jiahua Fine Biologi- cal Products Co. METHODS: ①OA model was established in rabbits of the two groups. Each rabbit was anesthetized intravenously with 1.0 mg/kg ketamine hy- drochloride and underwent unilateral anterior cruciate ligament transection. ②5 weeks after transection, Na-HA injection group rabbits received 0.3 mL of intra-articular 10 g/L Na-HA injection, once a week for 5 weeks. Ani mals in saline controlled group were treated with saline of the same vol ume. ③The rabbits were killed at week 10 after operation, general morphology and histopathological changes of articular cartilage degeneration of medial femoral condyle were evaluated (0 points as smooth articular surface with normal color and luster; 1 point as minimal fibrillation or a slight crevice and dark grey color of the surface; 2 points as erosion extending into superficial or middle layers of cartilage; 3 points as ulceration and erosion extending into the deep layers, and 4 points as denudation of cartilage, erosion extending to the sub-chondral bone). The mRNA expression of iNOS in cartilages was examined with reverse transcription-polymerase chain reaction (RT-PCR). MAIN OUTCOME MEASURES: ①Observation of articular cartilage degeneration of medial femoral condyle, ②observation of articular cartilage degeneration of medial femoral condyle at light microscopic level, ③expression of iNOS in cartilages of each group. RESULTS: A total of 16 clean healthy rabbits entered the result analysis. ① Result of articular cartilage degeneration of medial femoral condyle: Pathological change of articular surface of femoral condyle was observed under anatomic microscope. Cartilage degradation in experimental group was significantly less severe than that in saline control group. ②Histological changes of articular cartilage degeneration of medial femoral condyle at light microscopic level: The Na-HA injection group showed cartilage changes: Membrane of cartilage presented denaturalization and abscission. Chondrocytes of superficial zone presented denaturalization, necrosis, turbulence and erosion. Animals treated with saline showed denaturalization, necrosis and disorder of chondrocytes, ulceration penetrating into the middle or deep zone of the cartilage. New hyperplasia of capillary vessels and fibroblasts were more obvious. Proliferation of fibrous tissue appeared at the bottom of ulcer. ③Expression of iNOS in cartilages of two group:The gene expression of iNOS in cartilage of Na-HA injection group was(1.09±0.18) and the expression of saline control group was (1.26±0.23). Nosignificant difference of iNOS expression was found between the two groups(P ＞ 0.05).CONCLUSION: Intra-articular injection of Na-HA has protective and repairing effect on cartilage with early OA and can significantly reduce the severity of cartilage degradation during early stage of traumatic OA. Intra-articular injection of Na-HA does not down-regulated iNOS expression in cartilage.

Objective To study the effects of carboxymethylated chitosan (CMCS) to nitric oxide (NO)-induced apoptosis on rat chondrocytes,and explore p38MAPK signal transduction pathway in the process and its mechanism.Methods The rat articular cartilage cells were cultured in vitro,collagen type-2 (collagen-2) immunohistochemical staining was used to identify the cartilage cells.The model of chondrocyte apoptosis was built by different concentrations of sodium nitroprusside (SNP) induction.The cells were divided into the control group,the SNP treated group SNP+CMCS treated group,and the SNP+p38 MAPK inhibitor SB203580 treated group.The apoptotic rate of chondrocytes was calculated by FCM,apoptotic nuclei was identified by Hoechst33342 stain,the mitochondrial membrane potential changes was detected by Rhodamine123 (Rho123) stain,the expression of p38 and p-p38 were detected by Western blotting analysis.Results 1-3 mmol/L SNP could induce chondrocyte apoptosis,the apoptotic rate was increased with the SNP increasing,the most obvious apoptosis was occurred in 3 mmol/L SNP treated chondrocytes,which was 69.8％ (P＜0.05).SNP could increase the nuclear fragmentation of chondrocytes,the cells with nuclear fragmentation was significantly higher than that in the control group.SNP could reduce mitochondrial membrane potential in chondrocytes,which decreased significantly compared with the control group.SNP could increase the p-p38 expression in chondrocytes,which was 4.3 times compared to the control group.CMCS of different concentrations could reduce the apoptotic rate of SNP-induced chondrocytes,which was 51.0％,29.9％ and 15.2％,which was decreased significantly (P＜0.05) when compared with 3 mmol/L SNP induced group,CMCS decreased the cells number of SNP-induced nuclear fragmentation.CMCS increased the mitochondrial membrane potential in SNP-induced chondrocytes.CMCS reduced the expression levels of p-p38 in SNP-induced chondrocytes.Conclusion CMCS has protective effect on SNP-induced apoptosis of chondrocytes.This process is completed by inhibiting the activity of p38 MAPK signal pathway.

Objective To investigate the recheck rule by investigating the coincidence rate of the results detected by the LabU‐Mat urine dry chemistry analyzer and the Urised tangible composition analyzer with the results detected by the microscope examina‐tion .Methods 1 040 urine specimens from children outpatients and children inpatients were collected .Firstly ,the specimens were analyzed by the LabUMat urine dry chemistry analyzer and the Urised tangible composition analyzer ,and then detected by using the microscopic examination for investigating the recheck rule of the routine analysis by the urine automatic analyzer ;the regulation was evaluated by the missed detection rate ,and then the recheck rule avoiding the missed diagnosis of abnormal renal function was also evaluated .Finally ,clinically verify the rules adopting 200 specimens to perform the clinical verification on this recheck rule .Results Among the specimens used for researching the recheck rule ,the specimens of positive microscope examination results accounted for 58 .65% ,the specimens of negative results accounted for 41 .35% .In the positive detection specimens ,the specimens of RBC positive were the majority ,accounting for 50% ,the specimens of WBC positive accounted for 23 .08% and the specimens of CAST positive accounted for 7 .69% .The coincidence rate of the set rule was 87 .5% and the missed detection rate was 2 .9% .In conduc‐ting the verification on the recheck rule by 200 urine specimens ,the coincidence rate was 89 .52% and the missed detection rate was 2 .4% .Conclusion When the detection results of occult blood(BLD) ,WBC(LEU) and protein(PRO) by the dry chemistry analyzer and the detection results of RBC ,WBC ,CAST by the tangible composition analyzer are inconsistent or the differences among them are beyond 2 grades of differential ,the recheck by the microscopic examination should be performed .

OBJECTIVETo investigate the effects of Pyrroloquinoline quinine (PQQ) on hydrogen peroxide-induced apoptosis of Schwann cells (SCs) and its mechanism.

METHODSSCs were isolated and cultured in vitro, and identified by S-100 immunofluorescence staining. The cultured SCs were divided into control group, hydrogen peroxide-treated group, hydrogen peroxide and PQQ treated groups. The intracellular superoxide dismutase (SOD) and malondialdehyde (MDA) content was detected; the apoptotic rate of SCs induced by hydrogen peroxide was determined by flow cytometry assay. The Hoechst33342 staining was used to detect the nuclear fragmentation and apoptotic nuclear condensation of SCs; the Rhodamine123 staining was used to detect the changes of mitochondrial membrane potential in SCs, the Western blot analysis was used to detect the expression of Bcl-2 in hydrogen peroxide induced SCs.

RESULTSThe SOD activity was significantly decreased and MDA level was increased in H2O2 induced SCs (P < 0.05), after addition of PQQ, the SOD content increased and MDA content decreased (P < 0.05). Flow cytometry results showed that the early apoptotic rate was 58.8% in H2O2 induced SCs, which has significant difference compared with the control group (P < 0.05), after addition of 10, 50, 100 nmol/L PQQ, the apoptotic rates were reduced to 33.7%, 18.7%, 3.9% respectively, showing significantly different with injured group (P < 0.05). Hoechst 33342 staining showed that H2O2 induced SCs had typical morphological characteristics, such as uptake of nuclear chromatin, nuclear shrinkage, nuclear fragmentation phenomenon. The proportion of apoptotic cells after PQQ treatment reduced. Rhodamine staining results showed that the H2O2 induced mitochondrial membrane potential reduction in SCs, which was reversed by addition of PQQ. Western blot analysis showed that the expression of Bcl-2 was decreased in H2O2 induced SCs, while it increased significantly after addition of PQQ (P < 0.05).

CONCLUSIONPQQ has a protective effect on oxidative stress-induced apoptosis of SCs.

Apoptosis ; drug effects ; Benzimidazoles ; Cell Nucleus ; drug effects ; DNA Fragmentation ; Fluorescent Dyes ; Humans ; Hydrogen Peroxide ; pharmacology ; Malondialdehyde ; metabolism ; Oxidants ; pharmacology ; Oxidative Stress ; Pyrroles ; pharmacology ; Quinine ; pharmacology ; Quinolines ; pharmacology ; Schwann Cells ; cytology ; drug effects ; Superoxide Dismutase ; metabolism

Objective To investigate the possibility of plastic and cosmetic operation incorporating epicanthoplasty with the double-eyelid procedure in one stage to achieve a more subtle scar in epicanthus patients with single eyelid. Methods After designing incision line of the epicanthic flap, double-eyelid line and the lower eyelid incision curved line, we cut the medial canthus skin transversely to reach a new medial canthus point, incised the part of front angle of medial palpebral ligamentum longitudinally and fixed the end point of front angle of medial palpebral ligamentum to nasal dorsum fascia firmly. Double eyelid operation was routinely performed to remove redundant small triangle skin as well as some patchy of orbicularis on the new medial canthus point. Along with eyelid edge about 1 to 2 mm to eyelash, the temporal side was cut with curved line till its disappearance by eyelid's "cat ear". The lower eyelid flap was separated downward and the superfluous flap and some patchy of orbicularis cut. The skin was su-tured to make postoperative scar hidden, blepharophimosis increased and the fold disappeared. Results 46 eyes (23 cases) were operated and satisfactory aesthetic results were obtained. Palpebral fissure was enlarged to 2 to 4 mm with epicanthic scar disappearance and formation of double-eyelid. Conclusions This is a simple and effective procedure with hidden epicanthic scar and the double-eyelid blepharoplasty could be performed simultaneously. Most patients receive satisfactory results during the 0.5 to 2 years' follow-up period except 2 cases with mild proliferation of epicanthus in half a year. It is especially suitable to correct the severe epicanthus palpebralis or epieanthus tarsalis.

Objective To investigate the optimized parameters of dye (SYBR Green Ⅰ) realtime fluorescent polymerase chain reaction (RF-PCR) for detecting αβT lymphocyte clones in the peripheral blood and its application in monitoring specific T cell clone in the peripheral blood mononuclear cells (PBMC) from patients with chronic hepatitis B (CHB). Methods The total RNA was extracted from the PBMC of six healthy donors, and was reversely transcripted into cDNA. Then the cDNA was amplified using RF-PCR with the primers specific for T cell receptor β viable region (TCRBV) gene families as upstream primers and the primer for T cell receptor (TCR) β constant region (TCRBC) as downstream primer. The annealing temperature,concentration of primers and the total number of cycles were comparatively analyzed. The optimized PCR was performed to investigate the 24 TCRBV gene families from 12 patients with CHB, and the PCR products were monitored by melting curve analysis, and the clone expansion of peripheral blood T cell was detected by peak-motif of melting curve analysis. Results The optimized annealing temperature, final premier concentration,the number of cycles were 60.6 ℃, 0.5 μmol/L and 40 cycles, respectively. The begin temperature for melting curve analysis was better as 80 ℃ compared to 75 ℃. There was mono-peak on melting peak chart for TCRBV gene families in PBMC from patients with CHB, and PCR products of the single peak were determined as monoclonal T cell by sequencing. Conclusions The optimized reaction parameters of RF-PCR for monitoring 24 TCRBV gene families are determined. The melting peak chart could be used to monitor the clone expansion of the peripheral lymphocytes and to detect the clone-specific T cells in the peripheral blood from patients with CHB.

BACKGROUND:Recently, non-fusion technology representing as artificial cervical disc replacement continues to improve. On the basis of reconstruction of disc structure and function of involved segments, cervical spine structure of surgery area segment is significantly close to dynamic and static load stress distribution required by natural physiological systems. It effects are apparent in protecting intervertebral facet joints of degenerated segment and structure and function of the cervical spine of adjacent segments and in maintaining cervical dynamic stability, which presented obvious methodological strengths compared with segmental fusion technology.
OBJECTIVE:To evaluate the clinical outcomes of anterior cervical discectomy and fusion and Bryan artificial cervical disc replacement in the treatment of single-level cervical spondylotic myelopathy or radiculopathy.
METHODS:A total of 43 middle and old age patients with single-level cervical spondylotic myelopathy or radiculopathy, who were treated from March 2010 to March 2012, were enrol ed in this study. They were randomly assigned to anterior cervical discectomy and fusion group (fusion group) and Bryan artificial cervical disc replacement group. Range-of-motion of cervical overal and adjacent intervertebral area near the intervertebral space was observed with radiography. During fol ow-up, postoperative recovery of neurological function was evaluated using Japanese Orthopaedic Association scale, visual analog scale and neck disability index.
RESULTS AND CONCLUSION:None patients experienced complications of neurovascular injury during and after the surgery. Range-of-motion of postoperative overal cervical vertebra and adjacent joint was improved in the Bryan artificial cervical disc replacement group compared with the fusion group. Neurological function was apparently improved after surgery in each group. At 3 months after surgery, scores of Japanese Orthopaedic Association, visual analog scale and neck disability index were significantly improved in the Bryan artificial cervical disc replacement group compared with the fusion group (P<0.05). During final fol ow-up, there were significant differences in visual analog scale scores between the two groups. Japanese Orthopaedic Association scale score and neck disability index score were similar between the two groups. During fol ow-up, no prosthesis sinking, displacement or heterotopic ossification were detected. These data indicated that artificial cervical disc replacement could effectively keep the range of motion of cervical segments and protect disc degeneration of adjacent segment. Mid-term fol ow up obtained similar improvement of neurological function of fusion surgery. The moderate-term and short-term efficacies of non-fusion technology were better than fusion technology in the treatment of single-level cervical spondylopathy.

Objective To observe the effect of inhaled glucocorticoids combined with theophylline in smoking patients with bronchial asthma.Methods Seventy-three patients with bronchial asthma were enrolled in this study and they were divided into observation group (34 cases,smoker) and control group(39 cases,no-smoker).They all accepted inhalation of budesonide 200 μ g/suck,bis in die,plus aminophylline tablet (0.1 g,ter in die,per os) for 3 months.After treatment for 3 months,the basal control rate was compared between two groups.The levels of asthma control test (ACT) score,peak expiratory flow (PEF),the first second forced expiratory volume accounted for the percentage of prediction (FEV1％pred),eosinophil leukocyte (EOS) count and immunoglobulin E(IgE) in two groups before and after treatment for 3 months were compared too.Results After treatment for 3 months,the basal control rate in observation group was 88.24％ (30/34),in control group was 92.31％(36/39),and there was no significant difference(P＞ 0.05).After treatment for 3 months,the levels of ACT score,PEF and FEV1％pred in two groups were increased and the levels of EOS count and IgE were decreased than those before treatment,and there were significant difference (P ＜0.05).After treatment for 3 months,the levels of ACT score,PEF,FEV1％ pred,EOS count and IgE in observation group were (22.43 ± 2.64) scores,(292.52 ± 98.64) L/min,(74.87 ± 4.83)％,(270 ± 180) × 106/L,(68.25 ± 13.89) U/L,in control group were(22.81 ± 2.27) scores,(300.34 ± 100.45) L/min,(75.26 ± 5.04)％,(210 ± 170) × 106/L,(65.47 ± 11.28) U/L,and there were no significant differences (P ＞ 0.05).Conclusions Low dose inhaled glucocorticoids combined with theophylline in treatment smoking and non-smoking asthmatics patients are equal.It can improve asthma symptoms and lung function,improve hormone sensitive and reduce airway inflammation.