Synchronization isolation method for multiple types of cells from mouse liver / 中华肝脏病杂志

Objective: To explore a simple and feasible method for the isolation and purification of hepatocytes, hepatic stellate cells (HSC), and lymphocytes from mice. Methods: The cell suspension was obtained from male C57bl/6 mice by hepatic perfusion through the portal vein digestion method and then isolated and purified by discontinuous Percoll gradient centrifugation. Trypan blue exclusion was used to determine cell viability. Glycogen staining, cytokeratin 18, and transmission electron microscopy were used to identify hepatic cells. Immunofluorescence was used to detect α-smooth muscle actin combined with desmin in HSCs. Flow cytometry was used to analyze lymphocyte subsets in the liver. Results: After isolation and purification, about 2.7×10(7) hepatocytes, 5.7×10(5) HSCS, and 4.6×106 hepatic mononuclear cells were obtained from the liver of mice with a body weight of about 22g. The cell survival rate in each group was > 95%. Hepatocytes were apparent in glycogen deposited purple-red granules and cytokeratin 18. Electron microscopy showed that there were abundant organelles in hepatocytes and tight junctions between cells. HSC had expressed α-smooth muscle actin and desmin. Flow cytometry showed hepatic mononuclear cells, including lymphocyte subsets such as CD4, CD8, NKs, and NKTs. Conclusion: The hepatic perfusion through the portal vein digestion method can isolate multiple primary cells from the liver of mice at once and has the features of simplicity and efficiency.

Case-control study on remnant-preserving strategy for preservation and reconstruction of anterior cruciate ligament / 中国骨伤

OBJECTIVE@#To investigate and compare the clinical efficacies of remnant-preserving and remnant-non-preserving, remnant-non-preserving remnant segment preserving and remnant root preserving with anterior cruciate ligament reconstruction.@*METHODS@#From March 2014 to December 2017, 204 patients with anterior cruciate ligament (ACL) injuries were treated by single-bundle ACL reconstruction with hamstring tendon autograft. According to the different methods of remnant preservation, the procedures were divided into remnant segment preserving group (A), remnant root preserving group (B), and remnant-non-preserving group (C). There were 37 males and 39 femalesin group A aged from 16 to 43 years old with an average of (28.80±5.41) years old. The time from injury to operation ranged from 2 to 11 weeks with an average of (3.68±1.04) weeks. In group B, there were 39 males and 25 females aged from 18 to 41 years old with an average of (28.42±5.60) years old. The time from injury to operation ranged from 2 to 10 weeks with an average of (3.36±1.68) weeks. In group C, there were 37 males and 27 females aged from 18 to 43 years old with an average of (29.10±6.11) years old. The time from injury to operation ranged from 3 to 11 weeks with an average of (3.54±1.46) weeks. The range of motion (ROM) of the knee was used to assess the range of extension and flextion of the knee at pre-operation and 24 months after operation. Lysholm score and the international knee documentation committee (IKDC) score were used to assess the knee function. The differences among three procedures were judged by comparing among the three groups at 6, 12 and 24 months postoperatively.@*RESULTS@#All incisions got a one stage healing, and no complications, such as vascular injury, nerve damage and articular infect or the like, occurred. All the patients were followed up, and the follow-up duration of group A ranged from 24.00 to 45.96 months with a mean of (35.52±14.40) months;the follow up duration of group B ranged from 27.96 to 48.00 months with a mean of (37.56±10.68) month;and the follow up duration of group C ranged from 24.00 to 66.00 months with a mean of (37.08±13.44) month. There were no significant differences in follow up time among three groups (@*CONCLUSION@#Compared with remnant-non-preserving group, the residual tissue of anterior cruciate ligament is preserved, which is conducive to promote the healing and remodeling of tendon graft and accelerate the recovery of joint function. Proper fixation of residual tissue and restoration of its tension are the key factors affecting the postoperative efficacy.

Current diagnosis and treatment of posterior shoulder instability / 中国骨伤

The incidence of posterior instability of shoulder joint was significantly lower than that of anterior instability, but the clinical diagnosis and treatment was difficult, and the misdiagnosis and missed diagnosis rate were high. Its etiology, clinical manifestation and treatment strategy are totally different from the anterior instability. Therefore, the deep understanding of the anatomical structure around the shoulder joint, the mastery of the examination method, and the classification of the shoulder instability based on the anatomy and injury mode are of great importance to improve the accuracy of diagnosis. CT three-dimensional reconstruction is helpful to evaluate the defect of humeral head and glenoid bone, and MRA is helpful for the accurate diagnosis of posterior glenoid lip and joint capsule. The treatment was divided into conservative treatment and surgical treatment. Conservative treatment is recommended for muscular instability. Surgical treatment is recommended for traumatic and dysplastic instability. Different operative methods should be performed according to the injury of glenoid side or humeral head side. According to the condition of bone defect, soft tissue operation, bone grafting or osteotomy were performed to reconstruct the posterior stable structure of the glenoid injury; according to the area of the anterior bone defect, bone grafting or subscapular muscle packing were performed to the head of humerus defect. The former has the advantages of short learning curve and firm fixation, while the latter has the advantages of minimally invasive operation and the ability to observe the lesions from multiple angles and accurately control the location of bone masses. This paper summarizes the above problems.

Biomechanical study on different high-strength sutures and suture site for repairing posterior root tear of the medial meniscus / 中国骨伤

OBJECTIVE@#To compare biomechanical characteristic of different high-strength sutures and suture sites for repairing posterior root tear of the medial meniscus with modified Mason-Allen technique.@*METHODS@#Forty-eight specimen of medial meniscus of knee joint from fresh porcine (female, aged from 5 to 9 months with an average of 7 months) were chosen and established experimental model. The samples were divided into red zone fixation group and red-white zone fixation group according to suture sites, 24 in each group; and then were randomly divided into 3 subgroups which 8 in each group, and fixed with Ethibond suture, Ultrabraid suture and FiberWire suture, respectively. Biomechanical tests were performedon universal electromagnetic and mechanical testing machine. Each specimen was underwent 1 000 cyclic tests on the first time, then pull out test until failure. The maximum failure load, yield load, stiffness and displacement were analyzed.@*RESULTS@#All specimen were successfully completed biomechanical tests. The failure mode of Ethibond group was caused by suture fracture; 6 cases of Ultrabraid suture group was caused by suture fracture which belong to red zone fixation group, 10 cases were caused by suture pull out, which 2 cases belong to red zone fixation group, 8 cases belong to red-white zone fixation group;8 cases of FiberWire group was caused by suture pull-out. Biomechanical test showed that:(1)In terms of suture strength, comparison of the maximum failure load, yield load and stiffness showed that Ethibond suture group

Modified arthroscopic anterior approach for the treatment of shoulder Bankart lesion / 中国骨伤

OBJECTIVE@#To explore the effects of single anterior approach under shoulder arthroscopy in the treatment of Bankart injury, and to provide a scheme to overcome the difficulties and simplify the operation process.@*METHODS@#From July 2016 to January 2019, 62 patients with recurrent dislocation caused by Bankart leision were treated under shoulder arthroscopy, including 50 males and 12 females, ranging in age from 19 to 44 years old with an average age of (26.5±6.1) years old. There were 25 cases of left shoulder and 37 cases of right shoulder. The patients were divided into two groups according to the operation mode, 35 patients in the front modified single approach group (experimental group) and 27 patients in the traditional double approachgroup (control group). The results of the operation were evaluated by assessing the preoperative and postoperative ASES scores of the shoulder joint, and the time of the approach establishment and the overall operation process was recorded to evaluate whether the operation time of the experimental group was shortened.@*RESULTS@#Three patients were lost during the follow-up after operation, and except for one case of re-dislocation, all patients achieved satisfactory results without neurovascular injury and other complications. The patients were followed up before operation and 3, 6, 12 months after operation, and the ASES scores were significantly enhanced at the latest follow-up compared with those of before operation. The time of establishing surgical approach in the experimental group was (7.5±1.5) minutes, which was shorter than that of control group (13.7±1.2) minutes.@*CONCLUSION@#The improved technique of single-channel anterior approach has the advantages of reducing the surgical incision and shortening the operationtime while ensuring the quality of the operation, which is worthy of popularization and application.

Progress on graft and fixation options of arthroscopic anterior cruciate ligament reconstruction / 中国骨伤

Anterior cruciate ligament injury is one of the most common injuries of knee joint. Arthroscopic reconstruction of anterior cruciate ligament is the most commonly performed procedure. A variety type of fixation techniques are being used nowadays, such as interference screw, cortical suspension fixation and transfix, all of which can achieve good effect, with both advantages and disadvantages. However, the healing of reconstructed grafts is a complicated and long-term process, which can be affected by many factors. The mechanical properties of the grafts are also changed greatly in all stages of healing process, adding variables for the post-operative rehabilitation.

Hamstring tendon transplantation embedding reconstruction of medial patellofemoral ligament for the treatment of recurrent patellar dislocation / 中国骨伤

<p><b>OBJECTIVE</b>To explore the effectiveness of hamstring tendon graft embedding reconstruction of medial patellofemoral ligament for the treatment of recurrent dislocation of the patella.</p><p><b>METHODS</b>From March 2008 to June 2013,67 patients with recurrent dislocation of patella were treated, including 28 males, 39 females with an average age of 22 years ranging from 10 to 42 years old. The clinical manifestations involved knee joint instability,"giving way", sense of patellar dislocation, anterior knee pain. All 67 patients underwent arthroscopic hamstring tendon graft embedding reconstruction of the medial patellofemoral ligament. The curative effect were evaluated by preoperative and postoperative Lysholm score and Q angle changes.</p><p><b>RESULTS</b>All cases were followed up after operation and the mean follow-up time was(27.5±13.4) months(4 to 69 months). Postoperative incision were healed well and no patellar dislocation or subluxation occurred. Lysholm score improved from preoperative 76.35±2.86 to 82.71±2.29 postoperatively; Q angle decreased from preoperative (18.75±2.33)° to postoperative(13.28±1.75)°.</p><p><b>CONCLUSION</b>The method for the reconstruction of medial patellofemoral ligament can provide enough tension, patellar stability.imoroving the function of knee ioint.</p>

Arthroscopic patelloplasty and circumpatellar denervation for the treatment of patellofemoral osteoarthritis / 中国骨伤

<p><b>OBJECTIVE</b>To investigate surgical procedure and clinical effects for the treatment of patellofemoral osteoarthritis (PFOA) with arthroscopic patelloplasty and circumpatellar denervation.</p><p><b>METHODS</b>Totally 156 patients with PFOA (62 males and 94 females; aged from 45 to 61 years old with an average of 52) were treated from September 2011 and March 2013. The clinical manifestations included recurrent swelling and pains on knee-joint,and aggravated pains upon going up/down-stairs, squatting-down, or standing-up. PFOAs were treated with arthroscopic patelloplasty and circumpatellar denervation. The therapeutic effects before and after surgery were evaluated by Lysholm scores and Kujala scores. The therapeutic effects were graded by classification of cartilage injury degrees.</p><p><b>RESULTS</b>Totally 149 cases were successfully followed up for 14.8 months. The incisions were healed well and no complication were occurred. Lysholm score was improved from 73.29 preoperatively to 80.93 postoperatively and average Kajala score from 68.34 preoperatively to 76.48 after surgery. This procedure was highly effective for patients with I - III degree cartilage injury, while not for patients with IV degree cartilage injury.</p><p><b>CONCLUSION</b>For PFOA patients, arthroscopic patelloplasty and circumpatellar denervation is effective for significantly relieving anterior knee pains, improving knee-joint functions and quality of life, and deferring arthritic procession.</p>

Arthroscopic repair of meniscus injury with Fast-fix under local anesthesia / 中国骨伤

<p><b>OBJECTIVE</b>To evaluate the clinical outcome of arthroscopic repair method of meniscus injury with Fast-fix under local anesthesia.</p><p><b>METHODS</b>From October 2005 to September 2012,106 patients with meniscus injuries admitted into our - hospital were studied, including 74 males and 32 females, ranging in age from 13 to 71 years old, averaged 27.6 years old. The duration of the disease ranged from 15 days to 5 years. The main clinical manifestations included knee joint pain after exercise, joint locking, pressing pain of knee joint and positive McMurray signs. The MRI showed meniscus tear or degeneration. Arthroscopic repairing surgeries were performed with Fast-fix under local anesthesia. Each patient was assessed with VAS pain evaluation and Lysholm knee-joint score system before and after operation. All the patients were followed up more than 1 year.</p><p><b>RESULTS</b>One hundred and 2 patients were followed up by recording subjective symptoms, clinical examinations and questions naires for an average of 2.6 years (ranged, 1.1 to 8 years), and 4 patients lost follow-up. All the 102 patients had no anesthetic complications. Ninety-six patients had normal subjective symptom and clinical examinations. Four patients had a mild ache with activities,2 patients had moderate pain after activities with joint space pressing pain. VAS pain evaluation and Lysholm knee-joint score after operation both were much better than that before operation.</p><p><b>CONCLUSION</b>onclusion: Local anesthesia can provide nice circumstances for surgeries. Arthroscopic repair using Fast-fix is an idea method for meniscus injury, especially for the posterior horn tear of medial meniscus, which is simple and convenient with less complications, and satisfactory results.</p>

Etiological analysis and significance of anterior knee pain induced by gluteal muscles contracture / 中国骨伤

<p><b>OBJECTIVE</b>To explore causes of gluteal muscle contracture induced anterior knee pain and curative effect of arthroscopic release.</p><p><b>METHODS</b>From March 2002 to August 2013,36 patients with gluteal muscle contracture induced anterior knee pain were treated, including 15 males, 21 females, aged from 9 to 40 years old with an average (18.7±7.2) years old; the courses of diseases ranged from 4 to 30 years. The clinical manifestations involved limited to symmelia, positive Ober sign, buttocks touch contracture belts, knee and patella slide to lateral when doing squat activities. All patients were performed gluteal muscle contracture release under arthroscopic. Postoperative complications were observed, Kujala scoring before and after operation was used for compare curative effect.</p><p><b>RESULTS</b>All patients were followed up with an average of 29 months. The incision were healed well, and no complications were occurred. Postoperative Kujala score were improved more than preoperative.</p><p><b>CONCLUSION</b>Gluteal muscle contracture release could alleviate hypertension of lateral patella, and palys an important role in preventing patellofemoral arthritis.</p>

Effect of puerarin combined with felodipine on mRNA and protein expression of apelin and APJ in renovascular hypertensive rat / 中国中药杂志

<p><b>OBJECTIVE</b>To explore the effect of puerarin combined with felodipine on the mRNA and protein expression of apelin and APJ in renal tissue of renovascular hypertensive rat.</p><p><b>METHOD</b>Sixty-two Sprague-Dawley rats were used, of which 8 rats were randomly chosen as sham-operation group. The remaining rats were made for the rat model with renovascular hypertension. The renovascular hypertensive rats were randomly divided into 5 groups as follows: 4 groups which were treated with felodipine (0.8 mg x kg(-1) x d(-1)), puerarin (50 mg x kg(-1) x d(-1)), puerarin combined with felodipine (puerarin 25 mg x kg(-1) x d(-1) + felodipine 0.4 mg x kg(-1) x d(-1)) or captopril combined with felodipine (captopril 15 mg x kg(-1) x d(-1) x felodipine 0.4 mg x kg(-1) x d(-1)), and 1 group which was treated with distilled water. Drugs or distilled water were administered for 8 weeks. The expression of apelin and APJ mRNA and protein in ischemic and non-ischemic kidneys was assessed by RT-PCR or Western blot.</p><p><b>RESULT</b>Compared with sham-operation group, the expression of apelin mRNA and protein in ischemic and non-ischemic kidneys in model group was increased significantly (P < 0.01); the expression of APJ mRNA and protein in ischemic kidneys had no significance, while that in non-ischemic kidneys was decreased (P < 0. 01). Compared with model group, the expression of apelin mRNA and protein in ischemic and non-ischemic kidneys was decreased significantly in all drug-treated groups (P < 0.01); while that of APJ mRNA and protein in non-ischemic kidneys was upregulated (P < 0.01). Compared with felodipine group, the expression of apelin mRNA and protein in ischemic and non-ischemic kidneys was decreased (P < 0.01 or P < 0.05) in the group treated with both puerarin and felodipine; and the expression of APJ mRNA and protein in ischemic kidneys did not reach significant level, however, that was upregulated in non-ischemic kidneys (P < 0.01 or P < 0.05).</p><p><b>CONCLUSION</b>Puerarin downregulates the expression of apelin mRNA and protein in ischemic and non-ischemic kidneys, and upregulates that of APJ mRNA and protein in non-ischemic kidneys. Combination of puerarin and felodipine enhances the above-mentioned effects and shows no significant difference versus the combination of felodipine and captopril. The results suggest that puerarin regulates blood pressure and protects target organ through apelin/APJ pathway and that puerarin has synergetic effects with CCB.</p>

Association between single nucleotide polymorphisms of BARD 1 gene and susceptibility of early-onset breast cancer in Uygur women in Xinjiang / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the association between single nucleotide polymorphisms (SNPs) of BARD1 gene and susceptibility of early-onset breast cancer in Uygur women in Xinjiang.</p><p><b>METHODS</b>A case-control study was designed to explore the genotypes of Pro24Ser (C/T), Arg378Ser (G/C) and Val507Met (G/A) of BARD1 gene, detected by PCR-restriction fragment length polymorphism (PCR-RFLP) assay, in 144 early-onset breast cancer cases of Uygur women (≤ 40 years) and 136 cancer-free controls matched by age and ethnicity. The association between SNPs of BARD1 gene and risk of early-onset breast cancer in Uygur women was analyzed by unconditional logistic regression model.</p><p><b>RESULTS</b>Early age at menarche, late age at first pregnancy, and positive family history of cancer may be important risk factors of early-onset breast cancer in Uygur women in Xinjiang. The frequencies of genotypes of Pro24Ser (C/T), Arg378Ser (G/C) and Val507Met (G/A) of BARD1 gene showed significant differences between the cancer cases and cancer-free controls (P < 0.05). Compared with wild-type genotype Pro24Ser CC, it showed a lower incidence of early-onset breast cancer in Uygur women with variant genotypes of Pro24Ser TT (OR = 0.117, 95%CI = 0.058 - 0.236), and dominance-genotype CT+TT (OR = 0.279, 95%CI = 0.157 - 0.494), or Arg378Ser CC (OR = 0.348, 95%CI = 0.145 - 0.834) and Val507Met AA(OR = 0.359, 95%CI = 0.167 - 0.774). Furthermore, SNPS in three polymorphisms would have synergistic effects on the risk of breast cancer. In addition, the SNP-SNP interactions of dominance-genotypes (CT+TT, GC+CC and GA+AA) showed a 52.1% lower incidence of early-onset breast cancer in Uygur women (OR = 0.479, 95%CI = 0.230 - 0.995). Stratified analysis indicated that the protective effect of carrying T variant genotype (CT/TT) in Pro24Ser and carrying C variant genotype (GC/CC) in Arg378Ser were more evident in subjects with early age at menarche and negative family history of cancer. With an older menarche age, the protective effect was weaker.</p><p><b>CONCLUSIONS</b>SNPs of Pro24Ser(C/T), Arg378Ser (G/C) and Val507Met (G/A) of BARD1 gene are associated with significantly decreased risk of early-onset breast cancer in Uygur women in Xinjiang. Early age at menarche and negative family history of cancer can enhance the protective effect of mutant allele.</p>

Inhibition of CaMKⅡ? Expression by RNA Interference in PC12 / 中国生物工程杂志

To investigate the inhibitory effects of RNAi expression vector on CaMKⅡ? expression of PC12 cell lines. siRNA expression vectors were constructed to be targeted directly at CaMKⅡ? gene. The recombinants were transfected into PC12 cell line with liposome. The expression of CaMKⅡ? was detected by RT-PCR and Western blot. Expression vectors could reduce the expressions of CaMKⅡ? mRNA and protein in PC12, Compared with blank vector group, the former, the ratio of inhibition of the expression of CaMKⅡ? mRNA was 46.40% and 64.69%, in 48th hour and 72th hour, the ratio of inhibitory of the expression of CaMKⅡ? protein was 74.77% in 72th hour. RNAi expression vectors can effectively inhibit the expression of CaMKⅡ? in PC12.

External fixation and bone grafting for collapsed and comminuted distal radius fracture / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>To study the therapeutic effect of collapsed and comminuted distal radius fracture.</p><p><b>METHODS</b>Twenty-six patients with collapsed and comminuted distal radius fracture were hospitalized from July 1998 to June 2003. All fractures were treated by the methods of open reduction, sustained bone grafting and passing joint external fixator to restore the anatomic shape of distal radius.</p><p><b>RESULTS</b>All 26 cases were followed up, and the results showed that the fractures have been united radiographically. The joint surfaces were intact and there was no length discrepancy occurred in patient's radius. The average volar tilt was 6 to 15 degrees and the average ulnar tilt was 18 to 25 degrees. According to the Dieust criterion, 19 cases were rated as excellent and 7 as good.</p><p><b>CONCLUSIONS</b>The method that applying passing joint external fixator and bone grafting for the treatment of collapsed and comminuted distal radius fracture could maintain the stability of fracture and restore the length of radius and the intact of joint surface.</p>

The research actualities and developing trend of nitric oxide (NO) inhalation systems / 中国医疗器械杂志

This paper discusses the limitations of current NO inhalation systems, based on the research in collocation of NO, inspection of NO/NO2 and synchronous working of NO inhalation systems with ventilators. And then, the developing trend of NO inhalation systems is put forward here too.