Role of HBV DNA polymerase in mediating the immune escape of tumor cells in HBV-related hepatocellular carcinoma / 临床肝胆病杂志

ObjectiveTo determine whether HBV DNA polymerase is associated with T-cell failure and thus mediates the immune escape of HBV-related hepatocellular carcinoma （HCC） tumor cells， and to investigate the specific molecular mechanisms. MethodsLiver cancer cell lines Huh7 and HepG2 stably transfected with HBV DNA polymerase expression plasmid with Flag （Flag-HBV-P） and intercellular adhesion molecule-1 （ICAM1） were co-cultured with Jurkat cells， and MTT assay， qRT-PCR， and ELISA were used to measure Jurkat cell proliferation， activation （CD69 expression）， and secretion of the cytokine IFN-γ. RNA-seq was used to screen for differentially expressed immune-associated molecules between stably transfected cell lines and control cells， and mRNA half-life and protein half-life assays were used to determine the specific levels of the immune-associated molecules that were affected by HBV DNA polymerase. Related websites were used to predict the transcription factors that may bind to the promoter region of this immune-associated molecule， Western blot was used to verify the effect of transcription factors on the immune-associated molecule， and rescue experiment was used to determine whether HBV DNA polymerase affects the expression level of the immune-associated molecule through this transcription factor. The independent-samples t test was used for comparison between two groups. ResultsThe experimental group had significant reductions in Jurkat cell proliferation， activation， and cytokine secretion compared with the control group （all P<0.01）. Compared with the control group， the experimental group （Huh7 and HepG2 cell lines） had significant reductions in the mRNA and protein expression levels of ICAM1 （all P<0.01）. Website prediction identified the ICAM1 promoter and preliminarily highlighted NFKB1， RELA， and STAT3. Compared with the control group， the experimental group （Huh7 and HepG2 cell lines） had a significant reduction in the protein expression level of p65 （all P<0.01）. After p65 overexpression， there was a significant increase in the protein expression level of ICAM1， and after the expression of p65 was reduced， there was a significant reduction in the protein expression level of ICAM1 （all P<0.01）. In the rescue experiment， there was no significant difference in the protein expression level of ICAM1 between the control group and the experimental group after p65 overexpression （all P>0.05）. After the overexpression of ICAM1， there were no significant differences in the proliferation， activation， and cytokine secretion of Jurkat cells between the control group and the experimental group （Huh7 and HepG2 cell lines） （all P>0.05）. ConclusionHBV DNA polymerase downregulates the level of ICAM1 to mediate HCC immune escape by inhibiting the expression of p65 in NF-κB.

Analysis of clinical features of 78 infants with scabies / 中华皮肤科杂志

Objective:To analyze clinical features of 78 infants with scabies and their causes of misdiagnosis.Methods:A retrospective analysis was performed on infants aged < 6 months with confirmed scabies at Department of Dermatology in Zhengzhou Children′s Hospital, Jingjiang People′s Hospital, Tangdu Hospital of the Fourth Military Medical University, Second Affiliated Hospital of Soochow University and Henan Provincial People′s Hospital from January 1, 2016 to December 31, 2018. Then, epidemiological features, skin lesion characteristics, treatment and causes of misdiagnosis of infantile scabies were analyzed.Results:A total of 78 infants with scabies were collected. Their age of onset and duration from onset to diagnosis [ M ( P25, P75) ] were 8.5 (7, 12) and 4 (3.5, 5) weeks respectively. At the time of diagnosis, 45 (57.7%) patients showed lower body weight than the first quartile [ P25] of body weight of age- and gender-matched healthy peers, 40 (47.4%) had fussiness and irritation, and 68 (87.2%) had sleep disorders like night crying and increased frequency of night waking. Infantile scabies more frequently occurred in autumn (30 cases [38.5%]) and winter (22 cases [28.2%]) , and least frequently occurred in summer (8 cases [10.3%]) . In the case of 58 patients, there was at least 1 member with scabies at the same time, who had resided with the patients in their families for a long time; in the case of 12 patients, scabies was transmitted through previous contact with temporary residents with scabies in their families. Scabies lesions most commonly occurred on the chest and abdominal regions (80.8%) , followed by the limbs (76.9%) ; skin lesions were polymorphic, and lesions at different stages could coexist; the rashes mainly manifested as edematous red or non-edematous brown papules, blisters, papulovesicles and nodules, and some burrows could be characterized by an oval, linear, serpiginous, comma- or J-shaped appearance. All the patients had visited the clinic for 1 - 4 times with an average of 2.38 visits. Forty-eight (61.5%) patients initially visited non-dermatology departments, and 30 (38.5%) initially visited dermatological outpatient clinics. Incorrect diagnoses included infantile eczema, papular urticaria, impetigo, miliaria, prurigo, urticaria pigmentosa, infantile acropustulosis, herpes simplex and varicella. All the patients received topical sulfur 5% ointment. Nine (11.5%) patients experienced a sudden exacerbation of skin lesions after the topical treatment, and 20 (25.6%) needed 2 - 3 sessions of treatment. No recurrence was observed in all the patients at 2, 4 and 8 weeks after the end of treatment. Conclusions:Infantile scabies lesions are polymorphic, widely distributed, and easily misdiagnosed. To prevent misdiagnosis and improve the early diagnosis rate, a detailed clinical interrogation with clinical-epidemiological examination should be performed.

Expression of serum neutrophil gelatinase-associated lipocalin in children with psoriasis and its pathogenic mechanisms / 中华皮肤科杂志

Objective@#To explore the relationships of neutrophil gelatinase-associated lipocalin (NGAL) with severity of skin lesions in children with psoriasis and peripheral neutrophil count, and to evaluate in vitro effect of NGAL on expression of tumor necrosis factor-α (TNF-α) and interleukin-22 (IL-22) by a human immortalized keratinocyte cell line HaCaT.@*Methods@#From January 1st 2017 to December 31st 2018, 98 children who newly developed psoriasis were enrolled from Department of Dermatology of 6 hospitals in China, including 51 males and 47 females. Their age was 7.00 ± 2.99 years (range: 3-14 years) , and their course of disease was 7.4 ± 5.85 days (range: 3-28 days) . The serum level of NGAL was detected in all the patients before and two weeks after treatment, and the relationships of NGAL with psoriasis area and severity index (PASI) scores and peripheral neutrophil count were evaluated. Western blot analysis and reverse-transcription (RT) -PCR were performed to determine the protein and mRNA expression of TNF-α and IL-22 in HaCaT cells, respectively, after 12-hour treatment with NGAL at concentrations of 0 (control group) , 0.125, 0.25, 0.5, 1 mg/L. Statistical analysis was carried out with SPSS 16 software. by using t test and one-way analysis of variance.@*Results@#After 2-week treatment, the PASI score, neutrophil count and NGAL level in children with psoriasis significantly decreased (1.80 ± 1.19, [6.16 ± 0.76] × 109/L, 90.86 ± 0.75 μg/L, respectively) compared with those before the treatment (10.38 ± 3.42, [11.01 ± 2.85] × 109/L, 113.48 ± 21.26 μg/L, respectively; t = 31.42, 18.34, 16.37 respectively, all P < 0.001) . Before the treatment, the serum level of NGAL in the patients was positively correlated with the PASI score and peripheral neutrophil count (r = 0.918, 0.799 respectively, both P < 0.05) . The mRNA and protein expression of IL-22 in HaCaT cells significantly differed among these groups treated with different concentrations of NGAL (F = 176.31, 296.96 respectively, both P < 0.001) , so did the mRNA and protein expression of TNF-α (F = 193.28, 318.80 respectively, both P < 0.001) . Additionally, the protein and mRNA expression of IL-22 and TNF-α in HaCaT cells was significantly higher in the 0.125-, 0.25-, 0.5- and 1-mg/L NGAL group than in the control group (all P < 0.05) . The NGAL level was positively correlated with the protein and mRNA expression of TNF-α and IL-22 in HaCaT cells (all P < 0.05) .@*Conclusions@#The serum level of NGAL was high in children with psoriasis, and positively correlated with severity of skin lesions and peripheral neutrophil count. NGAL can upregulate the expression of TNF-α and IL-22 in HaCaT cells in vitro.

Expression of serum neutrophil gelatinase-associated lipocalin in children with psoriasis and its pathogenic mechanisms / 中华皮肤科杂志

Objective To explore the relationships of neutrophil gelatinase-associated lipocalin (NGAL) with severity of skin lesions in children with psoriasis and peripheral neutrophil count,and to evaluate in vitro effect of NGAL on expression of tumor necrosis factor-α (TNF-α) and interleukin-22 (IL-22) by a human immortalized keratinocyte cell line HaCaT.Methods From January 1st 2017 to December 31st 2018,98 children who newly developed psoriasis were enrolled from Department of Dermatology of 6 hospitals in China,including 51 males and 47 females.Their age was 7.00 ± 2.99 years (range:3-14 years),and their course of disease was 7.4 ± 5.85 days (range:3-28 days).The serum level of NGAL was detected in all the patients before and two weeks after treatment,and the relationships of NGAL with psoriasis area and severity index (PASI) scores and peripheral neutrophil count were evaluated.Western blot analysis and reverse-transcription (RT)-PCR were performed to determine the protein and mRNA expression of TNF-α and IL-22 in HaCaT cells,respectively,after 12-hour treatment with NGAL at concentrations of 0 (control group),0.125,0.25,0.5,1 mg/L.Statistical analysis was carried out with SPSS 16 software.by using t test and one-way analysis of variance.Results After 2-week treatment,the PASI score,neutrophil count and NGAL level in children with psoriasis significantly decreased (1.80 ± 1.19,[6.16 ± 0.76] × 109/L,90.86 ± 0.75 μ g/L,respectively) compared with those before the treatment (10.38 ± 3.42,[11.01 ± 2.85] × 109/L,113.48 ± 21.26 μ g/L,respectively;t =31.42,18.34,16.37 respectively,all P ＜ 0.001).Before the treatment,the serum level of NGAL in the patients was positively correlated with the PASI score and peripheral neutrophil count (r =0.918,0.799 respectively,both P ＜ 0.05).The mRNA and protein expression of IL-22 in HaCaT cells significantly differed among these groups treated with different concentrations of NGAL (F =176.31,296.96 respectively,both P ＜ 0.001),so did the mRNA and protein expression of TNF-α (F =193.28,318.80 respectively,both P ＜ 0.001).Additionally,the protein and mRNA expression of IL-22 and TNF-α in HaCaT cells was significantly higher in the 0.125-,0.25-,0.5-and 1-mg/L NGAL group than in the control group (all P ＜ 0.05).The NGAL level was positively correlated with the protein and mRNA expression of TNF-α and IL-22 in HaCaT cells (all P ＜ 0.05).Conclusions The serum level of NGAL was high in children with psoriasis,and positively correlated with severity of skin lesions and peripheral neutrophil count.NGAL can upregnlate the expression of TNF-α and IL-22 in HaCaT cells in vitro.

Alpha-mangostin affects the proliferation and apoptosis of gastric cancer cells by increasing autophagy / 西安交通大学学报(医学版)

Objective To investigate the possible molecular mechanism for alpha(α)-mangostin's inhibition of the proliferation and apoptosis of human gastric cancer cells.Methods Human gastric adenocarcinoma SGC7901 cell line was treated with α-mangostin.CCK8 method was used to measure the viability of SGC7901 cells.The effect of α-mangostin on apoptosis and cell cycle was determined by immune fluorescence and flow cytometry.The expression of the relevant proteins was detected using Western blot.The shapiro-wilk test was performed for evaluation of deviation from normality.Normally distributed data was analyzed with one-way ANOVA.Welch test was used in data with heterogeneity of variance and multiple compared by Games-Howell test after that.Results CCK8 results showed that cell viability differed significantly among groups treated with different concentrations of α-mangostin(10,15,20,25,and 30 μmol/L)(P＜0.05).QPCR data showed that the concentration of α-mangostin was positively correlated with mRNA level of LC 3 but not caspase protein(r=0.976,P＜0.05).In 15 μmol/L but not 10 μmol/L α-mangostin treatment system,the autophagy inhibitors 3-MA(10 μmol/L),bafilomycin A(10 μmol/L)and LY294002(10 μmol/L)could significantly alleviate α-mangostin's killing effect on SGC7901 cells(P＜0.05).Conclusion The anti-tumor effects of α-mangostin against human gastric adenocarcinoma cells in vitro can be partly attributed to apoptosis,autophagy and arresting cell phase.

Treatment of coronal shear fracture of the distal end of the humerus by the olecranon osteotomy approach / 中国骨伤

<p><b>OBJECTIVE</b>To explore the effectiveness of olecranon osteotomy approach for the treatment of coronal shear fracture of the distal end of the humerus.</p><p><b>METHODS</b>A retrospective analysis was made on the clinical data of 34 patients with the coronal shear fracture of the distal end of the humerus treated by the olecranon osteotomy approach from January 2005 to January 2013. Of 34 cases, there were 15 boys and 19 girls, aged from 17 years to 84 years (mean, 54.9±10.2 years); 18 patients had fractures on the left and 16 patients had fractures on the right. Fractures were classified according to the Bryan and Morrey classification united Mckee classification: type Iinjuries occurred in 10 cases, type IIinjuries in 5 cases, type III injuries in 10 cases and type IV injuries in 9 cases. The Mayo elbow functional scores were evaluated for analysis.</p><p><b>RESULTS</b>All the patients were followed up, and the duration ranged from 15 to 96 months, with a mean of (35.1±7.2) months. Average arc of motion was (132.1±11.2)° in flexion and (4.6±1.9)° in extension. The average Mayo score was 85.9±6.3(73 to 94 scores). Thirteen patients got an excellent result, 15 good and 6 poor. The average Mayo score was 88.6±3.7 in type Iinjuries, 85.8±4.6 in type IIinjuries, 81.8±5.8 in type III injuries and 87.5±9.1 in type IV injuries. There were no significant differences in outcomes.</p><p><b>CONCLUSIONS</b>The treatment for the coronal shear fracture of the distal end of the humerus by the olecranon osteotomy approach can achieve the satisfactory curative effect, maintain the reduction and improve the elbow function.</p>

Effects of acitretin on in vitro proliferation of HaCaT cells cultured in hypoxic condition and on expressions of hypoxia-inducible factor-1o and vascular endothelial growth factor / 中华皮肤科杂志

Objective To evaluate effects of acitretin on HaCaT cells cultured in hypoxic condition,and to preliminarily explore the possible therapeutic mechanisms of acitretin in psoriasis.Methods HaCaT cells were divided into several groups to be cultured in hypoxic condition with the presence of acitretin at concentrations of 10-5,10-6,10-7 and 10 8 mol/L respectively,with cells treated with dimethyl sulfoxide (DMSO) as DMSO control group and those receiving no treatment as blank control group.Cellular proliferative activity was evaluated by CCK-8 assay after 12-,24-and 36-hour hypoxic culture in vitro.The mRNA and protein expressions of hypoxia-inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) were determined by reverse transcription (RT)-PCR and Western-blot analysis,respectively,after 24-hour hypoxic culture.Results After 24-hour hypoxic culture,the cellular proliferation rate was inhibited by 13.31％ ± 1.15％,21.86％ ± 5.31％,32.05％ ± 2.99％ and 37.28％ ± 3.21％ in the 10 8-,10-7-,10-6-and 10-5-mol/L acitretin groups respectively.With the increase of culture duration and acitretin concentrations,the degree of inhibition on cellular proliferation increased gradually.Compared with the blank control group,the 10-5-mol/L acitretin group showed significantly decreased protein expression of HIF-1α (0.319 ± 0.180 vs.1.196 ± 0.088,P ＜0.05),as well as decreased mRNA and protein expressions of VEGF (mRNA:0.442 ± 0.090 vs.1.108 ± 0.073;protein:0.216 ± 0.066 vs.1.174 ± 0.186;both P ＜ 0.05).However,no significant difference was found in the mRNA expression of HIF-lα between the 10-5-mol/L acitretin group and blank control group.Conclusion Acitretin can suppress the in vitro proliferation of HaCaT cells cultured in hypoxic condition,and down-regulate the expressions of HIF-1α and VEGF proteins as well as VEGF mRNA.

Comparison of clinical effect on locking plate for proximal humeral fracture with or without application of inferomedial screws / 中国骨伤

<p><b>OBJECTIVE</b>To compare clinical outcomes of locking plate for proximal humeral fracture whether application of inferomedial screws.</p><p><b>METHODS</b>From January 2012 to July 2013, 46 patients with proximal humeral fracture underwent locking plates were retrospectively analyzed. There were 25 males and 21 females aged from 29 to 80 years old with an average of 55.1 years old. Among them, 25 patients were treated with inferomedial screws (support group), including 13 males and 12 females aged from 38 to 80 years old with an average of (55.8 ± 11.8) years old; 8 cases were part two fracture,10 cases were part three fracture and 7 cases were part four fracture according to Neer classification. Twenty-one patients were treated without inferomedial screws (non-support group), including 12 males and 9 females aged from 29 to 79 years old with an average of (54.2 ± 14.8)years old; 6 cases were part two fracture, 9 cases were part three fracture and 6 cases were part four fracture according to Neer classification. Operative time, fracture healing time and complications were observed and compared, Neer scoring of shoulder joint were used to evaluate clinical effect.</p><p><b>RESULTS</b>All patients were followed up from 12 to 41 months with an average of 15.6 months. Operative time and fracture healing time in support group was (1.6 ± 0.4) h and (3.0 ± 0.6) months, and (1.5 ± 0.4) h and (3.1 ± 0.6) months in non-support group, while there was no statistical difference in operative time and fracture healing time between two groups. There was significant differences in Neer score between support group (89.7± 4.9) and non-support group (83.1 ± 7.1). No complication occurred in support group,while 4 cases occurred complications in non-support group.</p><p><b>CONCLUSION</b>Locking plate with inferomedial screws for proximal humeral fracture has advantages of stable fixation, less complications, quick recovery of function and satisfied clinical effect.</p>

GC-MS analysis and cytotoxic activity of the supercritical extracts from roots and stems of Zanthoxylum nitidum / 中国中药杂志

The volatile components of roots and stems of Zanthoxylum nitidum were investigated by supercritical fluid carbon dioxide extraction (SFE-CO2) and gas chromatography-mass spectrometry(GC-MS). Thirty-one and fifty-one compounds were identified in the supercritical extracts from roots and stems of Z. nitidum, respectively, and total twenty-seven compounds were the common constituents. Among them, the major constituents in root and stem supercritical extracts were spathulenol (18.49 and 26.18%), n-hexadecanoic acid (14.24% and 12.79%), ar-tumerone (6.95% and 8.88%), oleic acid (8.39% and 5.71%) and hexanoic acid (4.39% and 7.78%). The in-vitro MTT assay showed that the volatile components of roots and stems of Z. nitidum did not exhibited any cytotoxic activity against human cancer Huh-7 and normal IEC-6 cells. These results indicated the same nature of the volatile constituents in the root and stem of Z. nitidum. This investigation may provide further evidence for expansion of medicinal parts of Z. nitidum.

A Microtus fortis protein, serum albumin, is a novel inhibitor of Schistosoma japonicum schistosomula

Schistosomiasis is an endemic parasite disease and praziquantel is the only drug currently in use to control this disease. Experimental and epidemiological evidence strongly suggests that Microtus fortis ( Mf ) is a naturally resistant vertebrate host of Schistosoma japonicum . In the present study, we found that Mf serum albumin ( Mf -albumin) and the conditioned medium of pcDNA3.1- Mf -albumin caused 46.2% and 38.7% schistosomula death rates in 96 h, respectively, which were significantly higher than that of the negative control (p < 0.05). We also found that mice injected with Mf -albumin had a 43.5% reduction in worm burden and a 48.1% reduction in liver eggs per gram (p < 0.05) in comparison to the control animals. To characterise the mechanisms involved in clearance, schistosomula were incubated with fluorescein isothiocyanate-labelled Mf -albumin and fluorescent enrichment effects were found in the gut lumen of schistosomula after 48 h of incubation. Next, digestive tract excretions from schistosomula were collected and the sensitivity of Mf -albumin to digestive tract excretions was evaluated. The results indicated that schistosomula digestive tract excretions showed indigestibility of Mf -albumin. The death of schistosomula could be partially attributed to the lack of digestion of Mf -albumin by digestive tract excretions during the development of the schistosomula stage. Therefore, these data indicate the potential of Mf -albumin as one of the major selective forces for schistosomiasis.

Efficacy analysis of coagulation disorders of patients with traumatic brain injury after plasma adjuvant therapy / 中华神经医学杂志

Objective To analyze the efficacy of plasma adjuvant therapy on coagulation disorders in patients with traumatic brain injury and its relation with the prognosis.Methods Prospective study was performed on patients with traumatic brain injury and coagulation disorders,admitted to our hospitals from January 2010 to June 2012; these patients were divided into conventional treatment group and plasma adjuvant therapy group.Cranial CT was performed and blood coagulation function was checked at admission,and then,coagulation function was re-checked again 3 days after therapy and cranial CT within 3 days.The Glasgow Outcome Scale (GOS) was marked 21 days after treatment.The improvement of coagulation disorders,secondary bleeding and prognosis were compared between the two groups.Results As compared with those in the conventional treatment group,the prothrombin time,partial thromboplastin time,secondary hemorrhage rate in the plasma adjuvant therapy group were significantly reduced (P＜0.05).The differences of D-dimer content and GOS scores between the two groups were not statistically significant (P＞0.05).Conclusion Plasma adjuvant therapy effectively improves the coagulation function,reduces the incidence of intracranial secondary bleeding,but can not obviously improve the prognosis of patients with traumatic brain injury.

Proliferation capacity and viability of adipose-derived stromal cells labeled with ultrasmall superparamagnetic particles of iron oxide / 中华神经医学杂志

Objective To label adipose-derived stromal cells (ADSCs) with different concentrations of ultrasmall superparamagnetic particles of iron oxide (USPIO) to investigate the biological characteristics of ADSCs treated with these USPIO,and determine the optimal concentration of USPIO in labeling ADSCs in vitro. Methods USPIO with different concentrations were prepared,and the particles were endocytosed by ADSCs generated from rat adipose tissue. Eight groups (negative control group,blank control group,and 11.25,22.5,45,90,135 and 180 μg/mL treatment groups) were chosen. Labeling efficiency and cellular uptake were analyzed by Prussian blue staining. Meanwhile,proliferation capacity and viability of ADSCs were evaluated by Alamar blue assay and Cell Counting kit-8. Results ADSCs could be effectively labeled with USPIO: approximately 95％ ADSCs were labeled when they were incubated with USPIO for 24 h under the concentration of USPIO was 45 μg/mL;and approximately 100％ ADSCs were labeled when the concentration of USPIO was 90 μg/mL and above. The CCK-8 and Alamar blue tests showed that USPIO of different concentrations (11.25-90 μg/mL) had little influence on cell growth viability,and no significant difference was noted between each 2 concentration groups (P＞0.05). In a word, 45-90 μg/mL USPIO were the optimal choice for transplantion of ADSCs in vivo. Conclusion ADSCs from the adipose tissue can be effectively labeled with USPIO with minimal effect on cell proliferation and viability.

Treatment of high-risk medulloblastoma by three dimensional conformal radiotherapy and intrathecal injection followed by systemic chemotherapy / 中华神经医学杂志

Objective To summarize the clinical efficacy of three dimensional conformal radiotherapy with concomitant intrathecal injection followed by systemic chemotherapy in the treatment of high-risk medulloblastoma (MB). Methods The clinical data of 20 patients with high-risk MB,admitted to our hospital from January 2005 to January 2008 and certified postoperatively by pathological examination, were retrospectively analyzed in our study; 15 of them were found tumors in the cerebrospinal fluid and MRI indicated 5 were with nodular cultivation in the spinal cord.Three-dimensional conformal radiation therapy was done for all the patients with the same techniques:28-36 Gy to the whole cranialspinal axis followed by a boost of 18 Gy. All the patients received intrathecal injection with cytarabine and amethopterin concomitantly followed by systemic chemotherapy with nimustine and temozolomide. The treatment efficacy was concluded. Results The mean follow-up was 48.5 months.Three months after treatment,the complete remission (CR) was achieved in 18 (90.0％),and partial remission in 1 (5.0％),and stabilization in 1 (5.0％).Four patients were failed of treatment and died; 1 had recurrence in the region,and 3 had spinal cord planting progress.The median survival time was 53 months; and 3-y overall survival and 3-y disease-free survival were 80.0％ and 75.5％,respectively.The major side effects occurred in hematological system and digestive system.No grade Ⅳ bone marrow suppression was noted. Conclusion Three-dimensional conformal radiotherapy and intrathecal injection followed by systemic chemotherapy for high-risk MB is safe and feasible,enjoying good prognosis.

Construction of a mouse pDsRed2-N1-SDF-1alpha eukaryotic expression vector and its expression in mouse bone marrow mesenchymal stem cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct the eukaryotic expression vector pDsRed2-N1-SDF-1alpha and observe its expression in the mouse bone marrow mesenchymal stem cells.</p><p><b>METHOD</b>SDF-1alpha gene sequence with XhoI, EcoRI restriction enzyme cutting site was amplified from the total RNA of mouse smooth muscle cells by reverse transcription-polymerase chain reaction (RT-PCR) and inserted into the eukaryotic expression vector pDsRed2-N1 encoding red fluorescent protein gene, and the insertion was verified by endonuclease digestion and DNA sequencing. Mouse bone marrow mesenchymal stem cells identified with immunofluorescence assay for vimentin expression were transfected with the constructed plasmid pDsRed2-N1-SDF-1alpha, and the expression of sdf-1alpha was detected using immunofluorescence assay.</p><p><b>RESULTS</b>The DNA fragment amplified by PCR from the total RNA was identical to SDF-1alpha from the gene library, and an identical DNA fragment was also amplified from the recombinants. Sequence analysis confirmed the successful insertion of SDF-1alpha into the pDsRed2-N1 vector and the eukaryotic expression vector pDsRed2-N1-SDF-1alpha was successfully constructed. The cultured mouse bone marrow mesenchymal stem cells positive for vimentin protein showed SDF-1alpha expression 24 h after transfection with the recombinant vector.</p><p><b>CONCLUSION</b>The pDsRed2-N1-SDF-1alpha eukaryotic expression vector constructed is capable of expression of SDF-1alpha fusion protein in the mouse bone marrow mesenchymal stem cells.</p>

Effect of adipose-derived GABAergic neuron transplantation in treatment of Parkinson's disease in rats / 中华神经医学杂志

Objective To investigate the method for inducing OABAergic neurons from adipose-derived mesenchymal stem cells (ADSCs) and observe the effect of transplantation of these neurons in the treatment of parkinsonian rats. Methods ADSCs isolated from rat inguinal fat pads were digested with collagenase, cultured and passaged in vitro, from which neural stem cells were induced using the neural stem cell culture medium prepared by our institute and identified for the stem cell markers. The neural stem cells obtained were further induced using the GABAergic neuron culture medium. After identification for the marker GAD65, the GABAergic neurons or the neural stem cells were stereotaxically transplanted into the subthalamic nucleus of the Parkinsonian rats, and the behavioral changes of the rats were observed at 2, 4 and 8 weeks after the cell transplantation. Results The neural stem cells differentiated from the ADSCs expressed the stem cell markers including nestin and neuron-specific enolase. After the second induction, the cells were positive for GAD65 as identified by immunofluorescence staining. Four weeks after transplantation of the neural stem cells and GABAergic neurons into the subthalamic nucleus, the parkinsonian rats exhibited significantly improved rotational behavior induced by apomorphine, and the improvement was especially obvious in rats with GABAergic cell transplantation. Conclusion GABAergic neurons can be induced from the rat ADSCs and transplantation of these neurons into the subthalamic nucleus can produce obvious behavioral improvement in rat models of Parkinson disease.

A clinical study of NexusTM coils for endovascular occlusion of intracranial aneurysms / 中华神经医学杂志

Objective To analyze the effect of NexusTM coils for endovascular occlusion of intracranial aneurysms. Methods In 41 patients with intracranial aneurysms, endovascular occlusion of 43 aneurysms was performed using NexusTM coils. The follow-up data of the patients for 6 to 12 months were reviewed, and the imaging data from digital subtraction angiography (DSA), CT angiography (CTA) or magnetic resonance arthrography (MRA) alter the treatment were analyzed. Results In the 41 patients, 1 died, 1 had aneurysm recurrence, 3 had cerebral infarction, 1 showed ocular paralysis, and 2 developed hydrocephalus after the surgery. Evaluation with modified Rankin Scale showed grade 0 in 8 cases, grade 1 in 19 cases, grade 2 in 7 cases, grade 3 in 3 cases, grade 4 in 1 case, grade 5 in 1 ease and grade 6 in 1 case. Conclusion Endovascular embolization with NexusTM coils is effective for treatment of intracranial aneurysms especially in cases of small aneurysms and parent artery occlusion. Caution should be taken with the coil for endovascular occlusion of the neck of anterior and middle cerebral artery aneurysms with thin parent arteries, as the fibers in the coil may cause thrombosis and potential cerebral infarction.

Endovascular embolization treatment of cerebral arteriovenous malformation with Glubran (NBCA-MS) / 中华神经医学杂志

Objective To explore the clinical skills and curative efficacy of Glubran (NBCA-MS) embolization of cerebral arteriovenous malformations. Methods Ninety-six cases of cerebral arteriovenous malformations were treated by 187 endovascular embolizations with different concentrations of Italy GEM company's Glubran (NBCA-MS). Follow-up was performed to 96 cases of cerebral arteriovenous malformation with NBCA-MS embolization. Results During the follow-up in 96 cases, clinical symptoms completely disappeared in 92, were improved in 3 and reoccurred in 1. The malformation was totally got rid of in 44 of 48 cases DSA examinated, and the 80%-90% embolization was achieved in the rest who need further γ-knife therapy. In all cases, γ-knife therapy was performed in 32 cases after embolization, and among them 26 cases received DSA examination 1-2 years after γ-knife therapy, and 21 cases were proved to have no malformation. Conclusions The method ofendovascular emdolization with NBCA-MS is safe, reliable and effective to treat the cerebral arteriovenous malformation.

In vitro culture and identification of neural stem cells derived from the olfactory bulb of adult mice / 中华神经医学杂志

Objecfive To establish a method for in vitro culture and identification of neural stem cells(NSCs)derived from the olfactory bulb(OB)of adult mice and test the possibility of the OB as a new source of seed cells of adult NSCs. Methads NSCs were isolated from the OB of adult mice and cultured in serum-free medium.Clonal culture and BrdU incorporation assay were performed to assess the self-renewal and proliferative activities of the NSCs.Fluorescence immunocytochemistry was carried out to examine the expression of the NSC markers nestin and SOX2,neuronal marker Tujl,astrocyte marker GFAP and oligodendroeyte marker 04. Results NSCs possessing self-renewal and proliferative capacities were obtained from the OB of adult mice,and the cells grew in the form of floating neurospheres in the medium.The neurospheres consisted of cells were positive for NSC markers nestin and SOX2,which Were able to differentiate into Tuj1-positive neurons,GFAP-positive astrocytes and 04-positive oligodendrocytes. Conclusion NSCs are present in the OB of adult mice,and the NSCs isolated from the OB can proliferate and differentiate in vitro with obvious stem cell properties.suggesting the feasibility of using OB as anew source of adult NSCs.

Culture and identification of neural stem cells derived from the subventricular zone of adult mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish a method for culturing and identifying neural stem cells (NSCs) derived from the subventricular zone (SVZ) in adult mice.</p><p><b>METHODS</b>NSCs were isolated from the SVZ of adult mouse brain and cultured in serum-free medium. Cell cloning and BrdU incorporation were performed to identify the self-renewal and proliferative capacity of the NSCs. Fluorescence immunocytochemistry was used to examine the expressions of the NSC markers nestin and SOX2, neuronal marker Tuj1, astrocyte marker GFAP and oligodendrocyte marker NG2. The expressions of nestin and SOX2 were further examined by Western blotting and RT-PCR.</p><p><b>RESULTS</b>NSCs with self-renewal and proliferative capacity were obtained from the SVZ of adult mice and grown as floating neurospheres. The NSCs expressed nestin and SOX2 and could differentiated into Tuj1-positive neurons, GFAP-positive astrocytes and NG2-positive oligodendrocytes.</p><p><b>CONCLUSION</b>This method allows simple and stable culture of NSCs from the SVZ of adult mice.</p>

Application of virtual imaging technique in diagnosis of intracranial aneurysms / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the value of virtual imaging technique in diagnosis of intracranial aneurysms.</p><p><b>METHODS</b>Fifty-four cases of 54 intracranial aneurysm diagnosed by three-dimensional CT angiography (3D-CTA) examinations were enrolled in this study. Three-dimensional virtual images of the skull and cerebral vessels were acquired by three-dimensional reconstruction of the original CT images using the surgical planning system, and the location, size and shape of the aneurysms and their anatomical relationship with the adjacent tissues were observed and measured from several angles. All the patients underwent surgical planning and simulated surgical operations using the virtual surgical instruments available in the system.</p><p><b>RESULTS</b>All the 54 cases had successful three-dimensional virtual image reconstruction and the surgical planning operations. The virtual imaging system generated clear and vivid three-dimensional virtual images which clearly visualized the location and size of the aneurysms and their precise anatomical relations to the parent arteries and skull. This virtual reality imaging system also allowed simulation of simple surgical procedures.</p><p><b>CONCLUSION</b>The surgical planning system based on the virtual reality imaging can serve as a useful means to assist the diagnosis and provide precise imaging details of intracranial aneurysms.</p>