[Objective]To explore the effect of tacrolimus on expression of hepatocyte growth factor(HGF)in rat spinal cord following peripheral nerve injury.[Method]Fifty male rats were randomly divided into normal group,injury group and treatment group.Models of peripheral nerve injury were established by transection of bilateral sciatic nerve at 0.5 cm distal to piriform muscle.Then treatment group received subcutaneous injection of tacrolimus(1 mg/kg)at the back of the neck,while injury group received 0.9 % saline.The L4~6 spinal cords were harvested at various time points after surgery.Western blotting and immunofluorescence staining were used to detect the level and position of HGF in the spinal cord.[Result]HGF-positive neurons were located in anterior horn,intermediate zone and posterior horn of gray matter in normal spinal cord.There was no significant difference in the expressions of HGF between injury group and normal group,while the expression of HGF was significantly higher in treatment group than that in injury group at 7 and 14 days after surgery.[Conclusion]Peripheral nerve injury does not up-regulate the expression of HGF in spinal cord,while tacrolimus can induce high expression of endogenous HGF after injury to protect neurons and promote axonal outgrowth.

Objective To retrospectively evaluate the clinical effects of end-to-end suture in repairing acute closed Achilles tendon ruptures.Methods Twenty-eight consecutive patients(20 males and 8 females,aged from 19 to 48 years with a mean of 36.5) with acute closed Achilles tendon rupture admitted from Nov.2000 to Jun.2006 were involved in present study.All patients were produced to have complete Achilles tendon rupture by MRI,and they received surgical procedure within 4 days after injury.An incision of 1cm long was made medial to the tendon,sparing the lesser saphenous vein and sural nerve.The tendon stumps were minimally debrided and end-to-end sutured using the Kessler or Bunnell techniques.The patients were followed-up for 12 to 36 months(mean 20 months),the therapeutic effects were evaluated by Arner-Lindholm scoring system,and postoperative complications were documented during the follow-up period.Results Of the 28 patients,19 got excellent therapeutic effect and 9 got good as evaluated by Arner-Lindholm scoring system,the satisfactory rate was 100%.All the patients regained normal gait,and rehabilitative training was initiated 3 months after operation.The postoperative complications included superficial wound infection in one case and skin edge necrosis in one case,resulting in a 7.1% incidence of complication.No peripheral nerve injury or Achilles tendon re-rupture occurred during the follow-up period.Conclusions An excellent therapeutic effect and minimal postoperative complication may be expected with end-to-end suture for repairing acute closed Achilles tendon rupture.

To find a new source of seed cells for constructing tissue-engineered intervertebral disc, nucleus pulposus (NP) cells and mesenchymal stem cells (MSCs) were isolated from New Zealand white rabbits. The nucleus pulposus cells population was fluorescence-laelled and co-cultured with MSCs with or without direct contact. Morphological changes were observed every 12 h. Semi-quantitative reverse transcriptase-polymerase chain reaction was performed to assess the expression levels of Sox-9, aggreacan and type II collagen every 24 h after the co-culture. MSCs treated with direct contact rounded up and presented a ring-like appearance. The expression of marker genes was significantly increased when cells were co-cultured with direct contact for 24 h. No significant change was found after coculture without direct contact. Co-culture of NP cells and MSCs with direct contact is a reliable method for generating large amount of NP cells used for cell-based tissue engineering therapy.
Aggrecans/metabolism ; Cell Differentiation ; Cells, Cultured ; Coculture Techniques ; Collagen/metabolism ; Gene Expression ; Gene Expression Regulation ; High Mobility Group Proteins/metabolism ; Intervertebral Disk/*cytology ; Mesenchymal Stem Cells/*cytology ; Mesenchymal Stem Cells/metabolism ; Models, Biological ; Reverse Transcriptase Polymerase Chain Reaction ; SOX9 Transcription Factor ; Tissue Engineering/instrumentation ; Tissue Engineering/*methods ; Transcription Factors/metabolism

This study is to investigate the effect of FK506 on expression of hepatocyte growth factor (HGF) in rats' spinal cord following peripheral nerve injury and to elucidate the mechanisms for neuroprotective property of FK506. Fifty male rats were randomly divided into normal group, injury group and treatment group. Models of peripheral nerve injury were established by bilateral transection of sciatic nerve 0.5 cm distal to piriform muscle. Then the treatment group received subcutaneous injection of FK506 (1 mg/kg) at the back of neck, while the injury group was given 0.9% saline. The L(4-6) spinal cords were harvested at various time points after the surgery. Western blotting and immunofluorescent staining were used to detect the level and position of HGF in spinal cord. Immunofluorescent staining showed that HGF-positive neurons were located in anterior horn, intermediate zone and posterior horn of gray matter in normal spinal cord. Western blotting revealed that there was no significant difference in the expressions of HGF between the injury group and the normal group, while the expression of HGF was significantly higher in the treatment group than in the injury group 7 and 14 days after surgery. It is suggested that peripheral nerve injury does not result in up-regulation of the expression of HGF in spinal cord, while FK506 may induce high expression of endogenous HGF after injury thereby protecting neurons and promoting axonal outgrowth.
Cells, Cultured ; Gene Expression Regulation ; Hepatocyte Growth Factor/metabolism ; Immunosuppressive Agents/metabolism ; Immunosuppressive Agents/*pharmacology ; Microscopy, Fluorescence/methods ; Neurons/metabolism ; Peripheral Nervous System/*metabolism ; Sciatic Nerve/metabolism ; Spinal Cord/*cytology ; Spinal Cord/metabolism ; Spinal Cord Injuries/*drug therapy ; Tacrolimus/metabolism ; Tacrolimus/*pharmacology

Objective To investigate the clinical effect and safety of miconazole nitrate 1200 mg in treating vulvovaginal candidiasis(VVC).Methods An open,multicentre,non case control clinical trial was conducted in 568 patients suffering from VVC from Jul 1,2006 to Nov 30,2006.Routine examination,score of clinical symptoms and physical signs,mycetology test and safety evaluation were done in all patients before treatment,7-14 days after treatment and 30 days after treatment.Results Seven to fourteen days after treatment,563 patients could be followed and 323 patients(57.30k)were cured.The overall effective rate was 90.2%.The mycologic cure rate was 91.3%(514).Thirty days after treatment,480 patients could be followed and 411 patients(85.6%)were cured.The total effective rate was 96.0%.Mycologic cure rate was 92.3%(443/480).Adverse effect rate was 2.7%(15/563)and they were relieved without any treatment in one or two days.Conclusions Miconazole nitrate 1200 mg is effective in the treatment of WC,with good compliance and few adverse effects.Moreover,it can be accepted easily.

Objective:To characterize the injury to the ipsilateral ankle joint after low energy spiral fracture of the distal tibia.Methods:A retrospective study was conducted to analyze the 80 patients with distal tibial spiral fracture who had been treated at Department of Traumatic Orthopedics, Affiliated Hospital of Jining Medical University from March 2010 to March 2021. There were 61 males and 19 females with an age of (43.5±12.5) years. Their mean follow-up time was 67.0 (38.5, 90.0) months. All patients underwent preoperative X-ray examination, 64 ones preoperative CT examination, and 30 ones preoperative MRI examination. Of the 80 patients, 3 received conservative treatment with plaster external fixation, 3 closed reduction and intramedullary nail fixation, and 74 plate fixation. Statistically analyzed were incidence of posterior malleolus fracture, characteristics of posterior malleolus fracture lines, normal matching rate of the ipsilateral ankle joint, positive rate of intraoperative Cotton test or stress external rotation test of ipsilateral ankle joint, positive rates of passive pronation-external rotation and supination-external rotation stress tests during follow-up, incidences of short-term ankle pain (≤2 years) and medium-long term pain (>2 years) after operation, injuries to the anterior inferior tibiofibular ligament, the deep medial malleolus deltoid ligament and the posterior malleolus, and incidence of ankle injury.Results:The diagnostic rate for posterior malleolus fracture was 16.3% (13/80) on X-ray film, 60.9% (39/64) on CT and 76.7% (23/30) on MRI. 74.5% (35/47) of the posterior malleolus fracture lines opened on the lateral side. The normal matching rate of the ipsilateral ankle joint was 96.3% (77/80). The positive rates of intraoperative Cotton test and stress external rotation test were 34.8% (8/23) and 7.1% (1/14), respectively. The positive rates of passive pronation-external rotation and supination-external rotation stress tests during follow-up were 46.2% (12/26) and 34.6% (9/26). The incidences of postoperative short term (≤2 years) and medium-long term (>2 years) ankle pain were 37.7% (29/77) and 20.8% (16/77). MRI examination showed that the rates of injury to the anterior inferior tibiofibular ligament, deep medial malleolus deltoid ligament and posterior malleolus were 80.0% (24/30), 80.0% (24/30) and 76.7% (23/30). The incidence of ankle injury was 88.8% (71/80).Conclusions:It is highly probable that spiral fracture of the distal tibia is complicated with ipsilateral ankle injury. The medial malleolus, lateral malleolus, and posterior malleolus are prone to the following hidden injuries while the ankle joint is normally matched in the most cases: injury to the deep deltoid ligament in different degrees, rupture of the inferior tibiofibular anterior ligament and posterior malleolus fracture. Therefore, the ankle injury is likely to be missed in diagnosis. The secondary torsion injuries to the pronation-external rotation and supination-external rotation at the leg are likely to cause ipsilateral ankle injury.

Objective To compare the clinical significance of peripheral blood neutrophil to lymphocyte ratio (NLR) and platelet to lymphocyte ratio (PLR) in predicting prognosis of patients with hepatocellular carcinoma (HCC).Methods The clinical data of 661 patients with HCC were retrospectively analyzed.Routine peripheral blood test results were used to calculate the NLR and PLR,and the receiver operating characteristic (ROC) curves were drawn.Using the thresholds of NLR and PLR,the patients were divided into the low NLR group and the high NLR group,and the low PLR group and the high PLR group.Overall survival (OS) and disease free survival (DFS) were evaluated by the Kaplan-Meier method.Independent prognostic predictors were determined by the Cox proportional hazard model.Results The NLR and PLR thresholds were 2.790 and 99,respectively.Analysis of the ROC curves showed higher NLR and PLR were significantly associated with poorer OS and DFS (all P ＜ 0.05).Multivariate analysis showed that NLR was an independent risk factor of OS and DFS (both P ＜ 0.05).The results remained unchanged when the NLR was further analyzed by applying different cut-off values of 2.810 and 3.In subgroup analysis,NLR remained an independent factor of Barcelona Clinical Liver Cancer staging system (BCLC) 0/A/B (P ＜ 0.05 for all measurements).Conclusion An elevated preoperative NLR could be a better prognostic predictor for HCC patients in comparison with PLR,especially for BCLC 0/A/B patients.

To find a new source of seed cells for constructing tissue-engineered intervertebral disc, nucleus pulposus (NP) cells and mesenchymal stem cells (MSCs) were isolated from New Zealand white rabbits. The nuclcus pulposus cells population was fluorescence-laelled and co-cultured with MSCs with or without direct contact. Morphological changes were observed every 12 h. Semi-quantitaive reverse transcriptase-polymerase chain reaction was performed to assess the expression levels of Sox-9, aggreacan and type Ⅱ collagen every 24 h after the co-culture. MSCs treated with direct contact rounded up and presented a ring-like appearance. The expression of marker genes was significantly increased when cells were co-cultured with direct contact for 24 h. No significant change was found after coculture without direct contact. Co-culture of NP cells and MSCs with direct contact is a reliable method for generating large amount of NP cells used for cell-based tissue engineering therapy.

This study is to investigate the effect of FK506 on expression of hepatocyte growth factor (HGF) in rats' spinal cord following peripheral nerve injury and to elucidate the mechanisms for neuroprotective property of FKS06. Fifty male rats were randomly divided into normal group, injury group and treatment group. Models of peripheral nerve injury were established by bilateral transection of sciatic nerve 0.5 cm distal to piriform muscle. Then the treatment group received subcutaneons injection of FK506 (1 mg/kg) at the back of neck, while the injury group was given 0.9% saline. The L4-6 spinal cords were harvested at various time points after the surgery. Western blotting and immunofluorescent staining were used to detect the level and position of HGF in spinal cord. Lm munofluorescent staining showed that HGF-positive neurons were located in anterior horn, interme- diate zone and posterior horn of gray matter in normal spinal cord. Western blotting revealed that there was no significant difference in the expressions of HGF between the injury group and the normal group, while the expression of HGF was significantly higher in the treatment group than in the injury group 7 and 14 days after surgery. It is suggested that peripheral nerve injury does not result in up-regulation of the expression of HGF in spinal cord, while FK506 may induce high expression of endogenous HGF after injury thereby protecting neurons and promoting axonal outgrowth.

Objective To investigate the changes of cannabinoid 2 receptor(CB2R) expression at different time after epileptic status in epileptic childhood rats. Methods Healthy male SD rats (18-21 days old) were randomly divided into two groups:control group and epileptic model group. CB2R concentration in hippocampus of rats were tested at 2 h,24 h,14 d,21 d after entering the status epilepticus( status epilepsy, SE) by immunohistochemistry,PCR,Western blot. Results In control group,CB2R content of the hippo-campus brain gradually increased with age increasing. When the rats with the age of 35-42 days,CB2R con-tent gradually got stabilized. After status epilepticus for 2 h-14 d,CB2R content of hippocampus in epileptic model group was more than that in the control group. At the point of 21 d,CB2R content of hippocampus in the control group was more than that in epileptic model group. CB2R mRNA of epileptic model group at 2 hours point was more than that of control group (2. 062 vs 1. 878,P<0. 05). At 24 h and 14 d after SE,there were significant differences between two groups in CB2R mRNA respectively ( P <0. 05, respectively). At 21 d after SE,CB2R mRNA of control group was more than that of epileptic group (6. 018 vs 5. 938),but there was no significant difference between two groups (P >0.05). Conclusion CB2R presents high expression in childhood rat hippocampus suffering from status epilepticus, reaches a peak following with prolonged seizures,then gradually decreased.