Objective:To perform a prospective cohort study to identify individual susceptibility of glucocorticoid (GC) -associated osteonecrosis of the femoral head (GA-ONFH) and their clinical and genetic risk factors. Methods:The present prospective cohort study enrolled patients who received their first GC therapy between July 2015 and January 2018 at Zhongshan Hospital. All patients did not receive any GC treatment before enrollment. Further, they planned to start GC treatment with the dose (equivalent prednisone) of ≥30 mg/d, lasted ≥3 weeks, or pulse dose ≥200 mg/d, lasted ≥3 d. Blood samples were collected before GC treatment to evaluate bone metabolism and its released factors. Hip MRI was performed at the 1st, 3rd, 6th, 12th and 24th month to diagnose GA-ONFH. All patients were followed-up for ≥2 years. The endpoint was regarded as diagnosis of GA-ONFH or completion of 2 years follow-up. Lasso regression was performed to determine which clinical features were associated with GA-ONFH. A nested case-control sub-cohort (A, n=12) was established prospectively based on the main cohort by 1∶1 matching. Whole exome sequencing was performed to screen differential and functional candidate single nucleotide polymorphisms and insertion-deletions (SNP/InDels). Another sub-cohort (B, n=50) was constructed retrospectively in patients with GA-ONFH and non-ONFH patients received standard high dose GC treatment for more than two years. The candidate SNP/InDels were verified by Sanger sequencing based on the patients from sub-cohort B. Results:A total of 96 patients were enrolled of which 88 of them (32 males and 56 females, mean age 42.30 years) completed follow-up. Eight cases (9.1%) were diagnosed with GA-ONFH. The median time from the start of GC therapy to the diagnosis of ONFH was 53.00(34.00,13.50) days. The baseline characteristics, such as age, sex and body mass index, indicated no significant difference between the ONFH group and the non-ONFH group. The cumulative GC dose of the ONFH patients in the first month was higher than that of non-ONFH [32.74(29.55, 47.05) mg/kg vs. 24.00(21.10, 29.45) mg/kg, Z=-2.410, P=0.016]. However, there was no significant difference of patients who underwent pulse therapy (37.5% vs. 10.0%, adjusted χ 2=2.829, P=0.093). The ratio of serum apolipoprotein B/apolipoprotein A1 (ApoB/ApoA1) in patients with ONFH was higher than that in non-ONFH group before GC use [0.95(0.80, 1.50) vs. 0.70(0.60, 0.80), Z=-2.875, P=0.000]. Due to the multicollinearity, Lasso regression model was performed to reduce overfitting. All variables were included in the model. The results suggested that higher ApoB/ApoA1 ratio, lower serum β-c-terminal telopeptide (β-CTX) and higher cumulative GC dose in the first month were the top three risk factors of GA-ONFH. This model had an accuracy of 0.982 in internal validation. Seven differential candidate SNP/InDels were found by whole exome sequencing of sub-cohort A. We further verified these SNP/InDels in sub-cohort B. The patients with COLEC12 mutation (rs2305027, G1816A) were at risk of GA-ONFH ( OR=6.00, 95% CI: 1.17, 30.73). Conclusion:Higher first-month GC dose, lower serum β-CTX level before treatment, higher ApoB/ApoA1 ratio and COLEC12 mutation (rs2305027, G1816A) could increase the risk of GA-ONFH.

Objective To evaluate the clinical efficacy and safety of short-segment pedicle screw fixation combined with vertebroplasty for the treatment of thoracolumbar osteoporotic compression fractures.Methods A retrospective case control study was conducted on 63 patients with fresh thoracic or lumbar osteoporotie compression fractures who were surgically treated from January 2010 to December 2013.There were 26 males and 37 females,with age of 63-87 years [(76.3 ± 5.7) years].According to the surgical method,the patients were assigned to simple vertebroplasty group (Group A),short-segment pedicle screw fixation group (Group B),and short-segment pedicle screw fixation group combined with vertebroplasty group (Group C),with 21 cases in each group.Length of hospital stay,operation time,and blood loss were recorded.The visual analog scale (VAS),anterior vertebral body height,angle of the kyphotic deformity,and complications before operation,immediately after operation,3 months after operation,and at the last follow up were compared among three groups.Complications of each group were recorded.Results The hospital stay,operation time,and blood loss in Group C were significantly higher than those in Group A (P ＜ 0.05),but there were no significant differences between Group B and Group C (P ＞ 0.05),except for a longer operation time in Group C.The pre-operative VAS showed no significant difference among three groups (P ＞ 0.05).However,the mean VAS in Groups A,B and C at the last follow up were 1.0(0.0,2.0)points,1.0(0.0,2.0)points,0.0(0.0,1.0)points,respectively.The VAS in Group C was significantly lower than that in Group B or A (P ＜ 0.05).The mean anterior vertebral body height and angle of the kyphotic deformity in Group C had no significant difference from that in Group A or B before operation and immediately after operation (P ＞ 0.05).At the last follow up,the mean anterior vertebral body height and angle of the kyphotic deformity in Groups A,B and C were (68 ±14.7)％,(72.3 ±9.0)％,(81.5 ±5.6)％ and (10.6 ±3.9)°,(10.7 ±5.0)°,(7.4 ± 5.0) °,respectively.The loss of anterior vertebral body height and angle of the kyphotic deformity correction in Group C were significantly less than that in Group A or B (P ＜ 0.05).Superficial infection was found in Groups B (n =2) and C (n =1),and the infection was cured after antibiotic therapy and dress change.Bone cement leakage was found in Groups A (n =8) and C (n =5),with no nerve compression.Internal fixation failure was seen in Group B (n =3),and the implant was removed directly.Conclusion Short-segment pedicle screw fixation combined with vertebroplasty can effectively reduce the loss of anterior vertebral body height and angle of the kyphotic deformity and hence enhance the clinical efficacy.

Objective To knock out the Abcb1 gene of rat,and establish the Abcb1 humanized rat model based on the Abcb1 knock out rat.Methods The animal model was established using BAC and CRISPR/Cas9 technology,and was analyzed by PCR, RT-PCR and real-time PCR.Results Establishing a rat model expressing human Abcb1 stably by transfer the 153 kb BAC containing human Abcb1 promoter and cDNA into rat genome, and establishing the Abcb1 knock out rat at the same time.Establishing the Abcb1 humanized model by crossing these two strains together.The expression pattern of Abcb1 in Abcb1 humanized rat is different from the wild type rat.The Abcb1 humanized model express not only the human Abcb1 gene but also has similar expression pattern as human.Conclusions The Abcb1 knock out rat and the Abcb1 humanized rat were successfully established, and this model is close to human concerning about the drug metabolism related to Abcb1.

Objective:To evaluate the efficacy of anteromedial extensile approach in the treatment of complicated closed pilon fractures classified as type AO/OTA C2 or C3 . Methods:Twenty‐three patients ,who suffered from complicated closed pilon fractures of distal tibia(25 lesions) ,underwent open reduction and internal fixation with anteromedial extensile approach . Among the 23 patients , there were 18 males and 5 females , with mean age of (43 .7 ± 9 .0 ) years . According to AO classification ,there were 11 lesions of type C2 and 14 lesions of type C3 .Operating time ,blood loss ,time of fracture healing , situation of complications ,and American Orthopedic Foot&Ankle Society(AOFAS) score were recorded ,so as to analyze the safety ,the indication ,and the pros and cons of this approach . Results:The average operating time was (119 .6 ± 23 .6)min . The intraoperative blood loss was (168 ± 64 .4) mL .And the time of fracture healing was (12 .5 ± 5 .4)weeks .The AOFAS scores were 81 .9 ± 18 .2 and 79 .9 ± 19 .5 at 12 month and 24 month after surgery ,respectively .Four cases of complications were observed , and there was no patient with invalid internal fixation . Conclusion:During the surgical treatment of complicated closed pilon fractures ,the anteromedial extensile approach allows complete access to the medial and lateral column of distal tibia and the articular surface ,so that the incidence rate of soft tissue complications was low and the clinical efficacy was satisfactory .

Objective:To explore the accuracy of CT three dimensional reconstruction technique for the diagnosis of tibia shaft fracture accompanied by ankle injury .Methods:Both X‐ray examination and three dimensional CT scanning were conducted for the 117 patients with closed tibia shaft fracture ,who went to Zhongshan Hospital of Fudan University ,before surgery .The numbers of patients ,who were diagnosed with tibia shaft fracture accompanied by ankle injury ,were compared between the two examination methods .Results:Nineteen patients with tibia shaft fracture were diagnosed with accompanied posterior malleolar fracture by X‐ray examination before surgery . However ,34 patients were diagnosed with accompanied posterior malleolar fracture by CT three dimensional reconstruction images . Thus , 15 more patients were diagnosed by CT three dimensional reconstruction technique than that by X‐ray imaging .Conclusions:Comparing with X‐ray examination , three dimensional CT reconstruction technique could increase the diagnosis accuracy for posterior malleolar fracture associated with tibia shaft fracture and reduce the rate of missed diagnosis .

Objective To study the relationship of insulin receptor substrate-1 (Irs1) and metabolic disease, we generated Irs1 gene knockout rat by CRISPR/Cas9 system.Methods Two sgRNA targeting sites were designed for Irs1 targeting.The Cas9 and sgRNAs were transcribed by T7 RNA polymerase in vitro.Cas9 mRNA and sgRNA mixtures were pooled and microinjected into one-cell fertilized eggs of SD rats to generate rats with targeted mutation .Results Five rats with the mutations were detected with the efficiency of 83%.Conclusion The Irs1 gene knockout rats generated in this study can be transmitted by germline .

This study examined the ability of 1,2,3,4,6-penta-O-galloyl-β-D-glucose (β-PGG) to induce the expression of heme oxygenase-1 (HO-1) in the PC12 cells and its regulation in the PC12 cells. One week before treatment with the drug, nerve growth factor (NGF) was added to the cultures at a final concentration of 50 ng/mL to induce neuronal differentiation. After drug treatment, HO-1 gene transcription was analyzed by reverse transcription polymerase chain reaction (RT-PCR). Expression of HO-1 and NF-E2-related factor2 (Nrf2) and activation of extracellular signal-regulated kinase (ERK) and Akt were detected by Western blotting. The viability of the PC12 cells treated with different medicines was examined by MTT assay. The oxidative stress in the PC12 cells was evaluated qualitatively and quantitatively by DCFH-DA. The results showed that β-PGG up-regulated HO-1 expression and this increased expression provided neuroprotection against MPP(+)-induced oxidative injury. Moreover, β-PGG induced Nrf2 nuclear translocation, which was found to be upstream of β-PGG-induced HO-1 expression, and the activation of ERK and Akt, a pathway that is involved in β-PGG-induced Nrf2 nuclear translocation, HO-1 expression and neuroprotection. In conclusion, β-PGG up-regulates HO-1 expression by stimulating Nrf2 nuclear translocation in an ERK- and Akt-dependent manner, and HO-1 expression by β-PGG may provide the PC12 cells with an acquired antioxidant defense capacity to survive the oxidative stress.
Animals ; Cell Death ; drug effects ; genetics ; Cell Line, Tumor ; Heme Oxygenase-1 ; genetics ; Hydrolyzable Tannins ; pharmacology ; MAP Kinase Signaling System ; drug effects ; genetics ; PC12 Cells ; Piperidines ; adverse effects ; Proto-Oncogene Proteins c-akt ; genetics ; Pyrazoles ; adverse effects ; Rats

This study examined the ability of 1,2,3,4,6-penta-O-galloyl-β-D-glucose (β-PGG) to induce the expression of heme oxygenase-1 (HO-1) in the PC12 cells and its regulation in the PC12 cells. One week before treatment with the drug, nerve growth factor (NGF) was added to the cultures at a final concentration of 50 ng/mL to induce neuronal differentiation. After drug treatment, HO-1 gene transcription was analyzed by reverse transcription polymerase chain reaction (RT-PCR). Expression of HO-1 and NF-E2-related factor2 (Nrf2) and activation of extracellular signal-regulated kinase (ERK) and Akt were detected by Western blotting. The viability of the PC12 cells treated with different medicines was examined by MTT assay. The oxidative stress in the PC12 cells was evaluated qualitatively and quantitatively by DCFH-DA. The results showed that β-PGG up-regulated HO-1 expression and this increased expression provided neuroprotection against MPP(+)-induced oxidative injury. Moreover, β-PGG induced Nrf2 nuclear translocation, which was found to be upstream of β-PGG-induced HO-1 expression, and the activation of ERK and Akt, a pathway that is involved in β-PGG-induced Nrf2 nuclear translocation, HO-1 expression and neuroprotection. In conclusion, β-PGG up-regulates HO-1 expression by stimulating Nrf2 nuclear translocation in an ERK- and Akt-dependent manner, and HO-1 expression by β-PGG may provide the PC12 cells with an acquired antioxidant defense capacity to survive the oxidative stress.

Objective:To evaluate the quality of prostate hyperplasia related literature in acupuncture and moxibustion,and to compare the curative effect on prostate hyperplasia between acupuncture and moxibustion and western medicine.Methods:Retrieving Pubmed,Cochrane Library,CBM database,CNKI database Etc.to collect the literature of prostate hyperplasia of clinical randomized or quasi-randomized control trials of comparative study between western medicine and acupuncture treatment.The data was extracted independently by two valuers from literatures fitting the selection criteria.Cochrane evaluation manual 4.2.6 was used to evaluate quality,and RevMan 4.2.8 was used in statistical analysis.Results:A total of six randomized or quasi-randomized controlled trials (total 546 examples) were adopted.6 study adopted the total effective rate of evaluation indexes,Meta-analysis showed that there was a significant difference between acupuncture treatment group and western medicine group [merger RR (fixed effects model)=1.26,95%CI(1.15,1.37),Z=5.13,P