The variable heavy chain region (VH) genes of 3 untreated patients with B-cell chronic lymphocytic leukemia (B-CLL) were cloned and analyzed. The VH family used was VH3-11, VH3-72 and VH3-33. More than 2% difference from the corresponding germline gene was detected in all the 3 obtained potential functional genes (average 16.7). Mutation pattern analysis indicated evidence of antigen selective pressure observed in 1 of 3 cases. Our findings suggested that the tumor cells originate from post-GC cells.
Amino Acid Sequence ; Gene Rearrangement ; Immunoglobulin Variable Region/*genetics ; Leukemia, B-Cell, Chronic/*genetics ; Molecular Sequence Data ; *Mutation

Objective To investigate the expression of inflanmatory cytokines such as interlukin-1β (IL-1 β),interlukin-6 (IL-6),tumor necrosis factor-α (TNF-α) and intercelluar adhesion molecule-1 (ICAM-1),and explore their relationships with neuronal function score after fluid percussion brain injury in rats.Methods The experimental models were established in 48 adult rats.The water content of edematous brain and the expression of IL-Iβ,IL-6,TNF-α and ICAM-1 were measured with dry-wet measure,immunohistochemistry at 6 h,12 h,24 h,3 d,7 d after operation respectively.Results Compared with animals of shame operation (SO) group,neuronal function score decreased in TBI group from 6h (3.78±0.84),reached to the lowest level at 24h (2.65±0.32),and gradually increased at 3d (4.75±0.71).IL-6,IL-1β,TNF-α and ICAM-limmunoreactive expression increased from 6h,reached its maxmum at 24h,lasted to 3th day,and began to decrease at 7d.The linear regression analysis indicated that expression of IL-1β,IL-6,TNF-α and ICAM-1 had negative correlations with change of neuronal function score(r=-0.912,r=-0.892,r=-0.794,r=-0.887 respectively,P＜0.05).Conclusion IL-6,IL-1β,TNF-α and ICAM-1 expression is upregulated following fluid percussion injury in rats,and shows negative correlations with neuronal function score.The possible mechanisms is that inflammatory cytokines are involved in nerve function impairment by inducing secondary brain injury.

Objective To investigate the expression of Toll-like receptor 4 (TLR4),and explore its relationship with neurological function after fluid percussion brain injury in rats.Methods 56 adult rats were randomly divided into traumatic brain injury group(TBI group,n=48) and sham operation group(SO group,n=8).The experimental models were established.The water content of edematous brain and the expression of TLR4 were measured with dry-wet measure,immunohistochemistry and Western Blot at 1 h,6 h,12 h,24 h,3 d,7 d after shock respectively.Results Compared with SO group,neuronal function score decreased in TBI group from 6 h(3.86±0.42),reached to the lowest level at 24 h(2.65±0.32),and gradually rose at 3rd day (3.25±0.17).TLR4 immunoreactive expression increased from 6 h,reached its maxmum at 24 h,lasted to 3rd day,and then began to drop at 7th day.The linear regression analysis indicated that expression of TLR4 had negative correlation with change of neuronal function score (r 1 =-0.824,r w =-0.867,P＜0.05).Conclusion TLR4 expression is upregulated following fluid percussion injury in rats and involved in neurological function impairment by inducing secondary inflammatory brain injury.

Objective To investigate the influence of m4-1BBL on the anti-tumor effects induced by truncated human prostate specific membrane antigen (tPSMA) gene in mice. Methods A eukaryotic expression plasmid encoding tPSMA and m4-1BBL (pDC316-tPSMA-IRES-m4-1BBL), pDC316-tPSMA and pDC316 were constructed. C57BL/6 mice were vaccinated in the quadriceps femoris, respectively. The CTL activity of spleen cells from the immunized mice against prostate cancer RM-1-tPSMA was detected by CCK-8 kit in vitro. The tumor growth was then observed. Results The target cell specific cytotoxicity rate induced by pDC316-tPSMA-IRES-m4-1BBL was 42.6%, compared to 24.8% in the pDC316-tPSMA group and 10.8% in the pDC316 group. The difference was significant (P<0.05). The volume of tumor in the pDC316 group was 2657.4mm3 7 d after vaccination, compared to 1334.5 mm3 in the pDC316-tPSMA group, 9 d after vaccination. In the pDC316-tPSMA-IRES-m4-1BBL group, the tumor volume was 445.8 mm3, 12d after vaccination. The difference was significant (P<0.05). Conclusion Gene vaccines co-expressing tPSMA gene and m4-1BBL gene could significantly enhance anti-prostate cancer effects in mice.

OBJECTIVE: To observe the clinical effect of perforated bovine amnion combined with recombinant bovine basic fibroblast growth factor (rb-bFGF) on degree Ⅱ burn wounds.METHODS: A total of 43 patients with small and medium-size thermal burn were collected, and the area of testing wound was 1% -2%. The wounds with the same nature were divided into three groups: perforated bovine amnion (treatment group), bovine amnion (control 1 group), and vaseline gauze dressing (control 2 group). All the three groups combined with rb-bFGF. RESULTS: Compared with control 1 group (P < 0.01) and control 2 group (P < 0.05), the treatment group could obviously decrease the healing time of deep degree Ⅱ burn wounds. For superficial degree Ⅱ burn wounds, compared with the control 2 group, the treatment group could also decrease the healing time; however, there was no significant difference between treatment group and control 1 group (P > 0.05). Dressing was not changed frequently, and the pain was relieved. Rash or other adverse effects were not detected in the three groups.CONCLUSION: The combination of perforated bovine amnion and rb-bFGF can obviously promote the healing of burn wounds.

The variable heavy chain region (VH) genes of 3 untreated patients with B-cell chronic lymphocytic leukemia (B-CLL) were cloned and analyzed. The VH family used was VH3-11, VH3-72 and VH3-33. More than 2% difference from the corresponding germline gene was detected in all the 3 obtained potential functional genes (average 16.7). Mutation pattern analysis indicated evidence of antigen selective pressure observed in 1 of 3 cases. Our findings suggested that the tumor cells originate from post-GC cells.
Amino Acid Sequence ; Gene Rearrangement ; Humans ; Immunoglobulin Variable Region ; genetics ; Leukemia, Lymphocytic, Chronic, B-Cell ; genetics ; Molecular Sequence Data ; Mutation

PURPOSE: The purpose of this study is to construct a recombinant adenovirus vector carrying mouse 4-1BBL and observe its effects in dendritic cells. MATERIALS AND METHODS: Mouse 4-1BBL cDNA was taken from the plasmid pcDNA3-m4-1BBL and subcloned into adenovirus shuttle plasmid pAdTrack-CMV, and then transformed into competent BJ5183 with plasmid pAdEasy-1. After recombination in E. coli, Ad-4-1BBL was packaged and amplified in HEK 293 cells. The expression of 4-1BBL in Ad-4-1BBL-transfected mouse prostate cancer cell line RM-1 was detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. After the co-culture of dendritic cells (DCs) with Ad-4-1BBL-transfected RM-1 cells, interleukin (IL)-6 and IL-12 production were assessed by enzyme-linked immunosorbent assay (ELISA) and co-stimulatary moleculs (CD80 and CD86) on DCs were analyzed by flow cytometry. RESULTS: The levels of IL-6 (3,960 pg/mL) and IL-12 (249 pg/mL) production in Ad-m4-1BBL-pulsed DCs were more than those in none-pulsed DCs. The differences were statistically significant (p < 0.05). The expression of co-stimulatary molecules (CD80 and CD86) was up-regulated in Ad-m4-1BBL-pulsed DCs. CONCLUSION: The results indicated the recombinant mouse 4-1BBL can effectively activate DCs.

Objective:To explore the effect of calcium phosphate cement（CPC）scaffold loaded with emodin（EMO）on osteogenic activity of osteoblasts.Methods:The bone cement scaffold was prepared by mixing EMO powder and CPC powder（ratio 1∶9），adding citric acid and then was poured into polytetrafluoroethylene mold（EMO-CPC group）. A dose of 0.36 g CPC powder was mixed with citric acid and injected into the polytetrafluoroethylene mold（CPC group）. General morphology，setting time（initial setting time and final setting time），injection rate and compressive strength of stents were compared between the two groups. Primary osteoblasts were extracted and co-cultured with two sets of scaffolds. After co-culture for 3 days，their characterization was observed by scanning electron microscopy. Live/dead cell staining and 3-（4,5-dimethylthiazole-2）-2,5-diphenyltetrazolium bromide（MTT）colorimetric method were used to detect cell viability，toxicity and proliferation activity of scaffolds. Two sets of scaffolds were stained with immunofluorescence for osteopontin（OPN），and protein expression was observed under an inverted fluorescence microscope. After co-culture for 7 days，tetrazolium nitro blue/5-bromo-4-chloro- 3-indolyl-phosphate（NBT/BCIP）staining method was used for alkaline phosphatase（ALP）staining. After co-culture for 14 days，two sets of scaffolds were stained with Alizarin Red to detect their osteogenic activity.Results:Two sets of stents showed relatively smooth and flat topography under the scanning electron microscope. There were no significant differences in initial setting time，final setting time，injection rate and compressive strength of stents between two groups（ P > 0.05）. After co-culture for 3 days，the osteoblast clusters were adhered to the surface of the EMO-CPC scaffold，with good shape. Viable cell rate reached（98.2 ± 0.1）% in EMO-CPC group and（90.2% ± 0.1）% in CPC group（ P <0.05）. Cell proliferation activity in EMO-CPC group was stronger than that in CPC group（ P < 0.05）. OPN-specific staining showed that EMO-CPC group had stronger OPN protein fluorescence expression compared to CPC group. After co-culture for 7 days，expression of ALP in EMO-CPC group was higher than that in CPC group. After co-culture for 14 days，staining intensity of Alizarin Red in EMO-CPC group was more significant than that in CPC group. Conclusions:The EMO-CPC scaffold can provide a suitable environment for the growth of osteoblasts for it has better biocompatibility，cell proliferation and osteogenic activity than the CPC scaffold.

Objective To avoid complications associated with the complete Senning procedures and to assist right-heart hemodynamics, this study was undertaken to assess the risks and benefits of the double-switch operation using a hemi-Mustard atrial switch procedure and the bidirectional Glenn operation for congenitally corrected transposition of the great arteries. we fa-vor a modified atrial switch procedure. Methods Between January 2014 and March 2016, anatomic repair was achieved in 26 patients. A hemi-Mustard procedure was the atrial switch procedure for 26. 9％(7/26). The Rastelli-atrial switch procedure was performed in 5 patients( RAS) and the arterial-atrial switch procedure was performed in 2 patients( AAS) . Results There was no in-hospital death after anatomic repair. There were no late deaths in a median follow-up of 7-21 months. Glenn opera-tion complications were uncommon limited to the perioperative period, atrial baffle-related reoperations or sinus node dysfunc-tion had not been observed. Tricuspid regurgitation decreased to mild for 1 case and mild to moderate for 6 cases. Right ventri-cle-pulmonary artery conduit longevity was significantly improved. Conclusion We recentely describe an experience with the double-switch operation using a modified atrial switch procedure with favorable earlyterm results. The risks of the hemi-mustard and bidirectional Glenn operation are minimal and are limited to a well-defined patient subset. The benefits include reduced baffle-and sinus node-related complications, technical simplicity and inspiring results.

Objective:To explore influence on physical development of children aged 18 months from HIV-positive mothers for prevention mother to child transmission of HIV (PMTCT) in Guangxi Zhuang autonomous region, and provide evidence for the improvement PMTCT program.Methods:This retrospective case control study was conducted in 554 HIV negative infants aged 18 months whose HIV positive mothers had received PMTCT services reported through PMTCT system database from January 1, 2010 to December 31, 2017 and 1 109 healthy infants born in 2017, whose mothers were healthy, in Lingshan, Luzhai, and Hengxian counties, ranking top three counties with high HIV infection prevalence, in Guangxi. PMTCT data and physical development data such as height, weight and head circumference of children aged 18 months were collected. The physical dysplasia in the infants was defined as at least one of the three main indicators of height, weight and head circumference below the normal range.Results:The number of HIV-positive mother and their infants in the case group were 667 and 554 respectively, and the PMTCT rates were 91.15 % (608/667) and 96.57 % (535/554) respectively. HIV positive rate, mortality rate and mother to child transmission rate of the infants aged 18 months were 1.44 % (8/554), 3.07 % (17/554) and 1.91 % (8/418) respectively, and the physical examination results of the infants aged 18 months showed that the physical dysplasia rate was 30.51 % (169/554). Among the 1 109 infants in the control group, the physical dysplasia rate was 9.83 % (109/1 109). The difference between the case group and the control group was significant ( P<0.01). Conclusion:The PMTCT rates of HIV positive mother and their children were more than 90.00 %, respectively. However, poor physical development rate of infants aged 18 months were more than 30.00 %. The possible influence of PMTCT on physical development of the infants aged 18 months of HIV positive mother’s needs to be further studied.