Relationship of urinary pathogenic bacteria and stone composition in patients with infectious stones / 中华泌尿外科杂志

Objective:To study the relationship of pathogenic bacteria in midstream urine culture and stone composition of patients characteristics with infection stones.Methods:Between January 2016 and December 2020, 989 patients with infectious stones who attended Tsinghua Chang Gung Hospital, affiliated with Tsinghua University, for surgical treatment were enrolled in the study. There were 545 male and 444 female patients, with the mean age (48±14) years. The left and right side stones were 396 and 333, respectively. There were 260 bilateral stones, 264 single stones, 334 multiple stones, and 391 deer-stalker-shaped stones. The maximum diameter of stones was (33.4±26.5)mm, combined with diabetes in 109 cases and hypertension in 235 cases. Clean middle-urine was collected for bacterial culture, and intraoperative stone specimens were collected by percutaneous nephrolithotomy (PCNL). Personal characteristics of the patient such as gender, age, body mass index, clinical information such as stone size, location, comorbidities, results of urine culture and stone composition were recorded. The differences of infectious stone composition was analyzed between urease-producing, non-urease-producing bacteria.Results:Among the 989 patients with infectious stones, 259 were pure infectious stones, 131 were mixed infectious stones, and 599 were combined with infectious stone components. Urine cultures were positive in 627(63.4%) patients with infectious stones. The predominant urease-producing bacteria included Ureaplasma urealyticum(94 case), Proteus mirabilis(58 case), and Staphylococcus spp.(36 case). Pure infectious stones were common in Proteus mirabilis, while combined with infectious stone components were common in Ureaplasma urealyticum and Staphylococcus spp. The predominant non-urease-producing bacteria included Escherichia coli(175 case), Enterococcus spp.(76 case) and Streptococcus spp.(35 case). Escherichia coli commonly contained in infectious stone components and pure infectious stones, whereas Enterococcus spp. and Streptococcus spp. commonly contained in infectious stone components. Escherichia coli (61 case), Proteus mirabilis (44 case) and Enterococcus spp.(20 case) were the most common bacteria in 259 cases of pure infectious stones. Escherichia coli (36 case), Enterococcus spp. (14 case) and Ureaplasma urealyticum (10 case) were the most common bacteria in 131 cases of mixed infectious stones. The most common bacteria in 599 cases of combined infectious stones were Escherichia coli (78 case), Ureaplasma urealyticum (68 case) and Enterococcus spp. (42 case).Conclusions:Urease producing bacteria were not common in infectious stones. It was common for the Ureaplasma urealyticum in combined infectious stone components, while Escherichia coli was common in pure and combined infectious stone components.

Application value of right minimal invasive three-port technique of laparoscopic sleeve gastrec-tomy for the treatment of obesity / 中华消化外科杂志

Objective:To investigate the application value of right minimal invasive three-port technique of laparoscopic sleeve gastrectomy (RMIT-LSG) for the treatment of obesity.Methods:The retrospective and descriptive study was conducted. The clinical data of 66 obesity patients who underwent RMIT-LSG in the Sir Run Run Shaw Hospital of Zhejiang University School of Medicine from January to October 2021 were collected. There were 15 males and 51 females, aged 28.5(range, 16.0?54.0)years. The body mass index (BMI) of the 66 patients was (36.9±4.3)kg/m 2. There were 20 of the 66 patients combined with type 2 diabetes. Observation indicators: (1) surgical situations; (2) postoperative situations; (3) follow-up. Follow-up was conducted using outpatient examination or the WeChat to detect postoperative recovery of patients including body mass changing, BMI and complications 6 months after operation. The follow-up was up to December 2021. Measurement data with normal distribution were represented as Mean± SD. Measurement data with skewed distribution were represented as M(range). Count data were described as absolute numbers. Results:(1) Surgical situations. All the 66 patients underwent RMIT-LSG successfully, without conversion to laparotomy or changing surgical method. The operation time and the volume of intraoperative blood loss of the 66 patients were (132±22)minutes and (14±8)mL, respectively. (2) Postoperative situations. The time to postoperative initial out-of-bed activities, time to postoperative first flatus, time to postoperative initial water intake, time to postoperative initial liquid food intake and duration of postoperative hospital stay of the 66 patients were (15±6)hours, (1.80±0.60)days, (1.00±0.20)days, (2.00±0.20)days and (3.40±0.60)days, respectively. Of the 66 patients, one case underwent post-operative abdominal hemorrhage at postoperative day 1 and received a second surgery for hemostasis. The patient with postoperative abdominal hemorrhage and other 65 patients recovered well without gastroparesis, gastric fistula, abdominal infection and other complication. (3) Follow-up. All the 66 patients were followed up for 6(range, 1?11)months. All the 66 patients completed the postoperative scar photography at postoperative 1 month, and results of scar photography showed concealed scar with good cosmetic effects. Twenty-seven of the 66 patients were followed up for 6 months after operation, with the weight loss, percentage of weight loss and decrease of BMI were (42±7)kg, 34.8%±2.9%, (14.2±1.9)kg/m 2, respectively. None of the 66 patient had innutrition during the follow-up. Conclusion:The RMIT-LSG is safe and feasible for the treatment of obesity, with a good cosmetic effect of the wound.

A case report of perioperative coagulation factor Ⅷ replacement therapy for hemophilia A with left hydronephrosis and hemorrhage / 中华泌尿外科杂志

Hemophilia is a recessive inherited disorder linked to the X chromosome. On July 28, 2020, a case of hemophilia A with severe left kidney hydronephroses and acute left kidney hemorrhage was admitted to the Affiliated Hospital of Qinghai University. The patient had a family history of hemophilia A and activated partial thromboplastin before surgery. The APTT was 95.0s (reference value 22.7-31.8s), factor Ⅷ activity (FⅧ∶C) was 3.2% (reference value 70%-150%), and B-ultrasonic examination revealed severe hydronephroses in the left kidney, and the nephrogram indicated that the left kidney was absent of perfusion. On the 9th day of admission, the pain in the left lower back was aggravated than before. The CT scan of the whole abdomen revealed irregular patches with slightly higher density in the left kidney. Considering bleeding, through multidisciplinary consultation and perioperative infusion of human coagulation factor Ⅷ, the patient was underwent retroperitoneal laparoscopic left nephrectomy. There was no bleeding during and after operation. After 20 months of follow-up, the renal function and coagulation function were normal, and there were no symptoms of hematuria and low back pain.

The triglyceride-lowering effects of PCSK9 inhibitor differ in patients with different baseline triglyceride levels / 南方医科大学学报

OBJECTIVE@#To investigate the triglyceride (TG)-lowering effects of PCSK9 inhibitor in patients with in different baseline triglyceride levels.@*METHODS@#Between February, 2019 and March, 2020, a total of 59 patients were treated with PCSK9 inhibitor (Evolocumab) in 5 hospitals, including Nanfang Hospital, Guangdong Provincial People's Hospital, First Affiliated Hospital of Sun Yat-sen University, Foshan Nanhai District People's Hospital and Yulin First People's Hospital. According to baseline triglyceride levels, the patients were divided into normal TG group (< 1.70 mmol/L, =24), mild hypertriglyceridemia group (1.70-2.29 mmol/L, =11), moderate hypertriglyceridemia group (2.30-5.63 mmol/L, =13), and severe hypertriglyceridemia group (≥5.64 mmol/L, =11), and the changes in TG level after the treatment were compared among the 4 groups.@*RESULTS@#In the groups with normal and mildly elevated baseline TG level, the patients did not show significant changes in TG levels after the treatment. In patients with moderately and severely elevated baseline TG levels, treatment with PCSK9 inhibitor significantly reduced their TG levels ( < 0.005).@*CONCLUSIONS@#PCSK9 inhibitor has a significant TG-lowering effect in patients with moderate to severe hypertriglyceridemia but not in patients with only mildly elevated baseline TG level.

Clinical efficacy and experiences of laparoscopic hepatectomy for segment Ⅶ and Ⅷ liver tumors / 中华消化外科杂志

Objective To investigate the clinical efficacy and experiences of laparoscopic hepatectomy (LH) for segment Ⅶ and Ⅷ liver tumors.Methods The retrospective cross-sectional study was conducted.The clinicopathological data of 94 patients who underwent LH for segment lⅦ or Ⅷ liver tumors in the Sir Run Run Shaw Hospital Affiliated to Zhejiang University School of Medicine from June 2010 to August 2016 were collected.The operating space for operation was built under laparoscopy.According to liver cirrhosis grading,tumor size,adjacent relationship with major blood vessels and residual liver volume,non-anatomical and anatomical hepatectomies were selected by patients.During the operation,tumors were precisely pinpointed and plane of liver resection was determined,and then proper instruments of liver partition and techniques of hepatic inflow occlusion were selected.Observation indicators:(1) surgical and postoperative recovery situations;(2) postoperative pathological examination;(3) follow-up and survival situation.Follow-up using outpatient examination and telephone interview was performed to detect the patients' survival up to July 2017.Measurement data with normal distribution were represented as (x)±s.Measurement data with skewed distribution were described as M (interquartile range).Survival rate was caculated by the Kaplan-Meier method.Results (1) Surgical and postoperative recovery situations:all 94 patients received successful operations,without perioperative death,including 73undergoing non-anatomical hepatectomy and 21 undergoing anatomical hepatectomy.Fourteen patients had conversion to open surgery and 27 received hepatic inflow occlusion.The median operation time,median volume of intraoperative blood loss and cases with intraoperative blood transfusion were respectively 187.5 minutes (75.0minutes),200 mL (200 mL) and 15.Eighteen patients had postoperative complications,including 6 with pleural effusion,6 with abdominal effusion,1 with wound infection,1 with abdominal infection,1 with venous thrombosis,1 with bleeding,1 with coagulation disorders and 1 with hepatic insufficiency.Clavien-Dindo classification of complications:11,1,5 and 1 patients were detected in grade Ⅰ,lⅡ,Ⅲ and Ⅳ,respectively.All complications were improved by symptomatic treatment.The median duration of hospital stay was 7 days (6 days).(2) Postoperative pathological examination:results of tumor pathological examination showed that 45,5,9 and 35 patients were respectively confirmed as hepatocellular carcinoma,cholangiocarcinoma,metastatic hepatic carcinoma and benign liver tumor.(3) Follow-up and survival situation:59 patients with malignant tumors were followed up for 6.0-52.0 months,with a median time of 42.6 months.Postoperative 1-and 3-year overall survival rates of 59 patients with malignant tumors were 98.3％ and 84.7％,respectively.Conclusions LH for segment Ⅶ and Ⅷ liver tumors which is conducted in experienced medical center is safe and feasible,with definite effects.Building operating space for operation under laparoscopy,determining precise positioning of the tumor and plane of liver resection,and selecting proper instruments of liver partition and techniques of hepatic inflow occlusion are the key points of successful operation.

Efficacy of nicergline combined with hyperbaric oxygen treatment on vascular cognitive impairment / 中华老年医学杂志

Objective To investigate the efficacy of nicergoline combined with hyperbaric oxygen treatment on vascular cognitive impairment.Methods 120 patients with vascular cognitive impairment were randomly divided into 3 treatment groups:nicergoline,hyperbaric oxygen,and both treatment (n=40,each) for 8 weeks.Mini mental state examination (MMSE),Wechsler memory scale (WMS) and activities of daily living scale (ADL) were used as assessing items.The efficacy of treatment in each group was evaluated.Results There were no significant differences in clinical data among the three groups(x2=0.324,t=0.265 and 0.861,P=0.764,0.784 and 0.386).After 8 weeks of intervention,scores of MMSE,WMS and ADL were significantly improved in the combined treatment groups compared with pre-treatment (all P＜0.01),and the improvements were observed in nicergoline group (all P＜0.05),while only MMSE and ADL were improved in hyperbaric oxygen group (both P＜0.05).There were significant differences in scores of MMSE,WMS and ADL between combination therapy group and nicergoline group,hyperbaric oxygen group after treatment (all P＜0.05).But only MMSE had significant difference between niccrgoline group and hyperbaric oxygen group after treatment.No significant side effects were observed in the three groups.Conclusions Combination treatment of nicergline and hyperbaric oxygen has better efficacy on vascular cognitive impairment.It can markedly improve the activities of daily living and cognitive function without significant side effects,which is worthy of application in clinical medicine.

Application of calcitonin for the treatment of unstable intertrochanteric fractures in elderly patients / 中华骨科杂志

Objective To compare the clinical outcomes between proximal femoral nail antirotation (PFNA) combined with salmon calcitonin and PFNA only for the treatment of unstable intertrochanteric fractures in elderly patients.Methods From January 2009 to December 2011,120 elderly patients with intertrochanteric fracture were randomly divided into two groups:calcitonin group and control group.Patients in calcitonin group were treated with PFNA combined with salmon calcitonin,while patients in control group were treated with PFNA only.According to Evans-Jensen classification,60 patients in calcitonin group (28 males and 32 females,with an average age of 75.1 years) were divided into 20 cases of type Ⅱ A,32 cases of type Ⅱ B and 8 type Ⅲ.Sixty patients in control group (27 males and 33 females,with an average age of 74.9 years) were divided into 22 cases of type Ⅱ A,32 cases of type Ⅱ B and 6 type Ⅲ.Bone healing was assessed with X-ray and bone mineral density (BMD) was measured by dual energy X-ray absorptiometry.Harris hip and SF-12 score,complications,adverse effect of salmon calcitonin and subsequent fragility fractures were evaluated postoperatively.Results One-hundred and thirteen patients were followed up for at least 2 years.In 6 months after surgery,there were 4 cases of delayed healing in control group.However,all fractures were healed in 12 months after surgery.No significant difference was found between the two groups in BMD preoperatively.The changes in BMD were significantly different between the two groups in 6 months,1 year and 2 years after surgery.No significant difference was found between the two groups in Harris hip and 1-year SF-12 score while the 2-year SF-12 score was significantly different between the two groups.There was 1 patient in calcitonin group who suffered from subsequent fragility fracture in 3 months after surgery,while there was 6 patients in control group during 13 to 23 months postoperatively.Conclusion PFNA combined with salmon calcitonin achieves good effect for the treatment of unstable intertrochanteric fractures in elderly patients.

Silencing IDH-2 gene by siRNA-IDH-2 inhibits human small cell lung car-cinoma growth / 中国病理生理杂志

[ABSTRACT]AIM:Toinvestigatetheeffectofsilencingisocitratedehydrogenase2(IDH-2)genebysmallinter-fering RNA (siRNA) on the biological characteristics of human small cell lung cancer cell line NCI -H446.METHODS:IDH-2 expression was knocked down in human small cell lung cancer cell line NCI -H446 by siRNA-IDH-2.The expression level of IDH-2 was determined by real-time PCR and Western blotting .The cell proliferation was measured by CCK-8 as-say , the protein expression of MAPK p 42 was detected by Western blotting , and the cell cycle was analyzed by flow cytome-try.The migration was observed using Transwell cell migration system .BALB/c nude mice were subcutaneously injected on the back with NCI-H446 cells transfected with siRNA-IDH-2/negative control siRNA or non-transfected cells to study the tumor growth .RESULTS:siRNA-IDH-2 remarkably down-regulated the expression of IDH-2 and MAPK p42 in the NCI-H446 cells.siRNA-IDH-2 inhibited both the proliferation and migration abilities of NCI-H446 cells, and the cell cycle was arrested in S phase as compared with negative control group .Additionally, the volume of xenograft tumors in siRNA-IDH-2 group was significantly decreased as compared with control group .CONCLUSION:siRNA-IDH-2 down-regulates the expres-sion of IDH-2 in NCI-H446 cells, reduces the cell migration efficiency and inhibits the tumor growth in vitro and in vivo.

Clinical application of T cell enzyme-linked immunospot assay in extrapulmonary on rapidly diagnosis of patients with tuberculosis / 中国综合临床

Objective To investigate the diagnostic value of T cell enzyme-linked immunospot (T-SPOT.TB) assay on extrapulmonary tuberculosis patients.Methods Thirty patients suffered from extrapulmonary mycobacterium tuberculosis(MTB) infection and 30 with non-MTB infection were recruited this study.T-SPOT.TB assay was used to detect early secreting antigen target-6 (ESAT-6) and culture filtrate protein-10(CFP-10) specific T cells in blood samples.PPD skin test was also used.Results (1)The positive rate of MTB detected by T-SPOT.TB assay was 91.89％ (34/37),higher than that of un-tuberculosis group (6.67 ％ (2/30)),and the difference was significant (x2 =48.403,P ＜ 0.001).(2) The sensitivity,specificity,positive prospective value and negative prospective value of T-SPOT.TB assay were 91.89％,93.33％,94.44％ and 90.32％ respectively,better than those of PPD skin test (67.57％,56.67％,65.79％,58.62％),and the differences were markedly (x2 =6.773,10.756,9.392,8.031 respectively ; P =0.009,0.001,0.002,0.005 respectively).Meanwhile T-SPOT.TB assay has low agreement with means of PPD skin test(Kappa =0.311,x2 =6.801,P =0.009).Conclusion T-SPOT.TB assay has a higher sensitivity and specificity in the rapid diagnosis of extrapulmonary tuberculosis.Therefore,it is with great value and applicability as a screening test.

Association of serum decoy receptor 3 protein level with the clinicopathologic features of bladder transitional cell carcinoma / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the association of serum levels of decoy receptor 3(DcR3) protein and the clinicopathologic features of bladder transitional cell carcinoma.</p><p><b>METHODS</b>Enzyme-linked immunosorbent assay was used to examine the serum levels of DcR3 in patients with bladder transitional cell carcinoma for analysis of its association with the patients' age, gender, clinical stages and pathological classification.</p><p><b>RESULTS</b>The patients with bladder transitional cell carcinoma showed a significantly elevated serum level of DcR3 (183.43 ∓78.45 pg/m1) compared with the normal level (116.65∓97.43 pg/m1, P<0.05). The serum level of DcR3 in the patients showed close correlations with the TNM stage and pathological classification of the tumor (P<0.05) but not with the patients' age or gender (P>0.05).</p><p><b>CONCLUSIONS</b>In patients with bladder transitional cell carcinoma, a high serum level of DcR3 suggests a higher malignancy of the tumor.</p>

Isolation and culture of human bone marrow mesenchymal stem cells without lysing the erythrocytes / 中国综合临床

Objective To observe the effects of red blood cell lysis buffer on the isolation and culture of human bone marrow mesenchymal stem cells (hMSCs) in vitro.Methods Twenty-two bone marrow samples were randomly divided into 2 groups,including 11 samples of crushed red blood cell prepared with Tris-NH4Cl red blood cell lysis agent and 11 samples prepared without red blood cell lysis agent.The intervals from primary generation to 1,2 and 3 passages and the time of expansion to l07 cells were compared between the two groups.Results The times for P1,P2 and P3 passage was ( 9.3 ± 4.9 ) days vs.( 7.2 ± 1.0 ) days,( 14.4 ±4.7) days vs.( 14.5 ± 3.5 ) days,and ( 18.5 ± 5.0 ) days vs,( 20.1 ± 4.4 ) days,respectively,in crushed red blood cell group and non-crushed red blood cell group.The differences were not significant ( t =1.39,t =0.06,t =0.80,P ＞ O.05 ).The time for expansion to 107 cells in two groups was not significantly different ( t =0.80,P ＞0.05).Conclusion Tris-NH4Cl agent in red blood cell lysis has no significant effects on hMSCs isolation,culture and cell proliferation,which indicates erythrocyte lysing may be not an independent step for hMSCs isolation and expansion in vitro.

Clinic study of unilateral pedicle screw combined with contralateral translaminar facet screw fixation by percutaneous and interbody fusion to treat low lumbar vertebra diseases / 中华骨科杂志

Objective To investigate the feasibility and efficiency of unilateral pedicle screw combined with contralateral translaminar facet screw fixation by percutaneous and interbody fusion to treat low lumbar vertebra diseases. Methods Thirty patients with low lumbar vertebra diseases were entered into the study, including 8 males and 22 females with an average age of 53.7 years. All patients underwent discectomy, spinal canal decompression, cage implantation and lumbar fixation by unilateral pedicle screw combined with contralateral translaminar facet screw under gunsight guiding by percutaneous. Clinical outcomes were assed by JOA questionnaires before and after operation. Operative time, blood loss, and postoperative draiming were recorded. Radiological examination was obtained to assess position of translaminar facet screw.Results Mean operation time was 89 min with a blood loss of 285 ml. Position of translaminar facet screw grade Ⅰ were 24 cases, and grade 11 were 6. Mean follow-up was 22.5 months. 29 cases got bony fusion, and the fusion rate was 96.7%. There were no instability and evidence instrument failure during follow-up. The JOA grades improved from 13.0 preoperation to 25.2 at final follow-up, with the excellent and good rate of 72.5 %. Conclusion Unilateral pedicle screw combined with contralateral translaminar facet screw fixation by percutaneous and interbody fusion provide simple procedure, little trauma, forceful fixation, high fusion rate, and less complication, etc. Therefore, the surgical maneuver is a good choice for partial low lumbar vertebra diseases.

The value of serum ICTP in the early diagnosis, efficacy and prognosis of tumor bone metastasis / 中国综合临床

Objective To evaluate the serum pyridinoline cross-linked carboxyteminal telopeptide of type Ⅰ collagen ( ICTP) in the early diagnosis potency,efficacy and prognosis of tumor bone metastasis. Methods According to emission computed tomography(ECT) ,MRI and X-ray results,336 cases of tumor were divided into higher ICTP (5. 98 ± 1. 95μg/L ) than normal values. Twenty-two cases were identified bone metastasis through PET/CT examination. 26 cases were identified bone the effective cases decreased from( 13. 22 ± 4.65)μg/L (before treatment) to (7. 18 ±3. 54)μg/L (after treatment) (t = 10. 076,P = 0. 000). Conclusions Serum ICTP is helpful for the early diagnosis, screening and efficacy evaluation of tumor bone metastasis. It could be used for monitoring the occurrence of tumor bone metastasis and its prognosis.

Relationship Between Variation and Distribution of Differential Gene Sites of Hepatitis B Virus and Enzymatic Activity of Liver Function / 中华医院感染学杂志

0.05).CONCLUSIONS G1896A Variation principally distributes in HBeAg(-) group that expressed low level HBV DNA.A1762T Variation and HBeAg(+) haven′t obvious correlation.YMDD variation principally distributes in HBeAg(+) group that expressed high lever HBV DNA.YMDD variation initiates acute damage of liver cell.The variation of G1896A or A1762T may contribute to progressive damage of chronic liver disease.

The full sequence of intron 51 of dystrophin gene and its characteristic of sequence / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To finish the work of sequencing the full sequence of intron 51 of dystrophin gene and understand its characteristic of sequence.</p><p><b>METHODS</b>The whole intron 51 was sequenced by primer walking. The sequencing results were analyzed by repeat sequences, matrix attachment region (MAR) and topoisomerase II cleavage sites. The residue sequences, after removal of the repetitive sequences, were subjected to the analysis of CpG islands, promoter, open reading frame (ORF) and unidentified low copy repeat sequence.</p><p><b>RESULTS</b>The acquired intron 51 sequence was composed of 38725 bp. Repetitive sequences constituted 37.53% of total intron sequence. The overall G+C content of intron 51 was 36.34%. There are four potential MARs in intron 51. Three of them are clustered in the 12 kb region near exon 51. Numerous ORFs were found on both strands, but no homologues proteins were found in Genbank CDS transcriptional peptide, PDB, SwissProt, PIR and PRF databases.</p><p><b>CONCLUSION</b>The expansion of intron 7 over the last 120 million years was mainly the result of L1 insertion into intron 7, and not all of repetitive sequences are associated with chromosomal rearrangement. No sequence of functional significance was found in intron 51. The results suggest that the cluster of MARs may be associated with the instability of intron 51.</p>

A new point mutation on exon 2 of parkin gene in Parkinson's disease / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To detect the relationship between point mutations on exon 2 of parkin gene and sporadic early-onset Parkinson's disease.</p><p><b>METHODS</b>The point mutations on exon 2 of parkin gene were detected using polymerase chain reaction(PCR), agarose electrophoresis, single strand conformation polymorphism(SSCP), DNA sequencing and analysis of restrict enzyme in DNA of 60 Parkinson's disease patients with an onset age under 50 and 120 normal controls.</p><p><b>RESULTS</b>One homozygous mutation (G(237)-->C) on exon 2 was found by sequencing and verified by analysis of restrict enzyme, whereas no mutation was found in normal controls.</p><p><b>CONCLUSION</b>Point mutations on exon 2 of parkin gene are likely to be related to sporadic early-onset Parkinson's disease.</p>

Mutation Detection on Exon 1 and 2 of Parkin Gene in Sporadic Early-onset Parkinson's Disease / 中山大学学报(医学科学版)

【Objective】To study the relationship between mutations on exon 1,2 of parkin gene and sporadic early-onset Parkinson's disease.【Methods】The deletion and single strand mobility shift on exon 1 and 2 of parkin gene in peripheral white blood cell DNA were detected by using PCR,agarose electrophoresis,and SSCP techniques in 52 patients with sporadic early-onset (onset age≤50) Parkinson's disease.The exons with mobility shift on SSCP were sequenced.【Results】One deletion（1.9%） of exon 2,2 cases with single strand mobility shift（3.8%）on exon 1 and exon 2 respectively,one heterozygous mutation (T103C) on exon 1 and one homozygous mutation (G237C) on exon 2 were found by sequencing.【Conclusion】Mutations on exon 1 and 2 of parkin gene are likely to be related to sporadic early-onset Parkinson's disease.

Transplantation of bone marrow mesenchymal stem cells improves motor function in a DMD mouse model / 中国病理生理杂志

AIM: To observe amelioration of motor function in a Duchenne muscular dystrophy (DMD) mouse model ( dko mice) after transplantation of bone marrow mesenchymal stem cells (MSCs). METHODS: Passage fifth MSCs cultured in vitro were transplanted into dko mice by tail vein, motor functions of experimental mice and matched control mice, including traction, rotating rods, rotated wheel, upside down, turning over and walking (all were recorded by Sony digital camera) were tested 15 weeks after transplantation. The fluorescent expression of dystrophin and utrophin in gastrocnemius muscle tissue of dko mice was detected by SABC-Cy3, and average optical density of positive fibers was calculated. RESULTS: MSCs grew in colony over passage third, and there was low immunologic reaction by vein transplantation. There was dystrophin and utrophin fluorescent expression in sarcolemma of dko mice 15 weeks after transplantation, but no any fluorescent expression in controls. There was significant difference in fluorescent average optical density of positive fibers between two groups ( P

Differentiation of rat marrow-derived mesenchymal stem cells into myoid cells in vitro / 中国病理生理杂志

AIM:To study the differentiation of rat marrow-derived mesenchymal stem cells(MSCs) into myoid cells in vitro . METHODS:The MSCs of SD rat were cultured?passaged?induced and differentiated in vitro used by routine culture technique, and evaluated by FACScan flow cytometer,detected by immunohistochemistry and analyzed by Hitachi H-600 transmission electron microscope(TEM). RESULTS:FACScan shows that cells expressed the antigens of CD29 and CD44, not those of CD11b and CD45; cells show positive response of staining with desmin and myoglobin after processing by two compounds of 5-azacytidine and amphotericin B. There were stripform zone of myofilament without any organells beside the edge of membrane in myoid cell of induction and differentiation checked by TEM. CONCLUSION: The passaged cells were MSCs and the MSCs may have specific gene structures that can differentiate into myoid cells. The demethylation or hypomethylation may conduct by compounds of 5-azacytidine and amphotericin B, which could be involved in activating phenotype-specific genes to differentiate MSCs into myoid cells. There are good outlook on clinical treatment of illness of myatrophy using by MSCs.

Genotype evaluation, muscle histopathology and ultrastructure in a Duchenne muscular dystrophy animal model / 中国病理生理杂志

AIM: To evaluate the genotype, muscle histopathology and ultrastructure in dko mice. METHODS: Dystrophin/Utrophin-deficient double knockouts (dko) mice were obtained from university of Oxford, UK. Genotype of filial generation of heterozygote was evaluated by PCR-SSP. HE staining and fluorescent immunohistochemistry by SABC-Cy3 were used to detect striated muscle of dko mouse, and the muscle ultrastructure was observed by transmission electron microscope(TEM). RESULTS: In 112 filial generation mice, there were 28 mdx (25.0%), 26 dko (23.2%) and 58 heterozygote (51.8%), which coincided with the law of Mendelian inheritance. HE staining showed that the myocytes were not very uniform, there were phenomenon of round outline, centrally nucleated fibers, widening interspace, inflammatory cell infiltration and connective tissue proliferation in dko mice. There were no any immunofluorescent expression of dystrophin and utrophin in sarcolemma in dko mice. TEM showed sarcolemma breakage, separation and edema, and loose myofibril texture, inflammatory cell infiltration and connective tissue proliferation in dko mice. CONCLUSION: PCR-SSP is a very quick and accurate way for genotype evaluation of filial generation. The pathophysiology of dko mouse was very similar to Duchenne muscular dystrophy (DMD), and dko mouse is an ideal animal model for study of DMD clinical therapy.