Adrenal Glands ; Female ; Hematopoiesis, Extramedullary ; Humans ; Spherocytosis, Hereditary ; complications ; Young Adult

Objective To investigate a new surgical method by using pectoralis minor muscle for its partial transplantation to reconstruct the function of the opposite thumb anastomosed vessels and nerves. Methods The proposed method was evaluated by taking 20 cases of adult cadaveric thoraxes and hands,then compared with the morphology and dimension of both pectoralis minor muscle and palmer muscle to study the feasibility of the new method.Based on the observed data.We selected five suitable cases,in which the opposi- tion function was lost,and then applied the new operation on them with partial transplantation of pectoralis mi- nor muscle anastomosed vessels and nerves according to the results of anatomic study.The follow-up study was conducted to observe the functional recovery of opposition of the thumb.Results The main results can be summarized as follows.First,the anatomical position of pectoralis minor muscle was stable,and every peetoralis minor muscle was provided with self-sustaining artery,vein and nerve.The oppostition process of cadaveric hand succeeded after similar transplantation to clinic operation.Second,follow-up studies conducted 6 - 12 months after the operation showed that all five patients recovered fully.The muscle strength in all five cases re- covered to level four or higher.The shape of palm eminence was satisfactory.Conclusion The surgical method of pectoralis minor muscle transplantation for reconstructing the opposition function of the thumb was based on the clinical and anatomical application.The function of opposition of the thumb reached the satisfacto- ry requirement after the operation.So,the new surgical method can achieve better results than other existing operation methods.

OBJECTIVETo observe the role of green fluorescent protein (GFP) in tracing rhesus bone marrow stromal cells (rBMSCs) during tissue-engineered bone formation in vivo.

METHODSAd5.CMV-GFP was amplified by infecting QBI-293A cells, and the bone marrow was harvested from the ilium of adult male rhesus to obtain rBMSCs, which were cultured and passaged in vitro. GFP was transfected into the third-passage rBMSCs via adenovirus vector and the labeled cells were inoculated into absorbable HA scaffold and cultured for 3 days, with untransfected rBMSCs as control, before the cell-matrix compounds were implanted into the latissimus dorsi muscles of rhesus. Samples were harvested at 6 week and embedded in paraform, and ground sections of the bone tissue were prepared to observe green fluorescence under laser scanning confocal microscope. Propidium iodide staining of the sections was also performed for observation.

RESULTSThe rBMSCs grew well after GFP transfection, and green fluorescence could be seen 24 h after the transfection and became stronger till 48 h, with a positive transfection rate beyond 80%. Six weeks after cell implantation, the rBMSCs labeled by GFP-emitted green fluorescence were detected in the bone tissue under laser scanning confocal microscope.

CONCLUSIONGFP can effectively trace BMSCs during bone tissue engineering, and the transplanted BMSCs constitute the main source of bone-forming cells in bone tissue engineering.

Animals ; Bone Substitutes ; Cell Differentiation ; Cells, Cultured ; Green Fluorescent Proteins ; genetics ; metabolism ; Macaca mulatta ; Male ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Microscopy, Confocal ; Tissue Engineering ; methods ; Transfection

OBJECTIVETo investigate the effects of hepatitis B virus (HBV) in semen on human semen parameters and sperm DNA integrity.

METHODSWe detected HBV DNA in the semen samples of 153 HBsAg-seropositive patients by real-time fluorescence quantitative PCR and calculated the sperm nuclear DNA fragmentation index (DFI) by sperm chromatin dispersion (SCD) assay. We compared the semen parameters between the HBV DNA-positive group (A, n = 43) and HBV DNA-negative group (B, n = 110) and analyzed the correlation of sperm DFI with the number of HBV DNA copies in the semen.

RESULTSHBV DNA was detected in 43 (28.1%) of the 153 semen samples. No statistically significant differences were observed in age, semen volume and sperm concentration between groups A and B (P >0.05). Compared with group B, group A showed significantly decreased sperm viability ([58.0 +/- 18.8]% vs [51.4 +/-17.1]%, P<0.05), progressively motile sperm ([29.6 +/- 13.3]% vs [24.5 +/- 10.1]%, P<0.05), average straight-line velocity ([23.7 +/- 4.0] microm/s vs [19.9 +/- 4.5 ] microm/s, P<0.01) and average path velocity ([26.5 +/- 7.0] microm/s vs [23.4 +/- 5.3] microm/s, P<0.01), but remarkably decreased sperm DFI ([19.3 +/- 8.0]% vs [24.2 +/- 9.4]%, P<0.01). The number of HBV DNA copies in semen exhibited a significant positive correlation with sperm DFI (r = 0.819, P < 0.01).

CONCLUSIONHBV DNA in semen is not significantly associated with the number of sperm, but may affect sperm viability, velocity and DFI. There is a load-effect relationship between the number of HBV DNA copies in semen and sperm nuclear DNA integrity.

Adult ; DNA Fragmentation ; DNA, Viral ; isolation & purification ; Hepatitis B Surface Antigens ; blood ; Hepatitis B virus ; genetics ; Humans ; Male ; Semen Analysis ; Sperm Count ; Spermatozoa ; virology

OBJECTIVETo understand the characteristics of embB gene mutation of Mycobacterium tuberculosis (MTB) isolates from tuberculosis patients in Chongqing, and the value of embB306 as a molecular marker used to diagnose ethambutol (EMB)-resistant MTB strains.

METHODSDirect sequencing was used to analyze the polymorphism of embB mutation in 51 EMB-resistant MTB strains and 50 EMB-sensitive MTB strains. And diagnostic testing was used to evaluate the value of embB306 as a molecular marker of EMB -resistant MTB strains as compared with the traditional sensitivity test.

RESULTSAll 34 of 51 EMB-resistant strains (66.7%) and 3 of 51 EMB-sensitive strains (6%) had had embB306 mutation. The embB306 mutation rate in EMB-resistant strains coming from previously treated case was 87.5%, showing significantly higher than that from new cases (48.1%, P < 0.01); embB306 mutation rate was increased with the number of the resistant drugs; embB306 mutation serving as a marker to diagnose EMB-resistant MTB strains comparing with the traditional sensitivity test, had the rate of sensitivity = 66.7%, specificity = 94.0%, accuracy = 80.2% and Youden index = 60.7%.

CONCLUSIONembB306 mutation should be the main mechanism of MTB resistance to EMB in Chongqing, showing an association with the history of the treated and numbers of the resistant drugs. embB306 mutation should be a good marker to diagnose EMB-resistant MTB strains.

China ; DNA Mutational Analysis ; DNA, Bacterial ; genetics ; Genes, Bacterial ; Humans ; Mutation ; Mycobacterium tuberculosis ; drug effects ; genetics ; isolation & purification ; Pentosyltransferases ; genetics ; Tuberculosis, Multidrug-Resistant ; microbiology

To express and secrete native HBscFv (anti-HBsAg single-chain Fv) in P. pastoris, HBscFv was amplified from plasmid pGEM-HBscFv, and then sub-cloned into expression vector pPICZalphaA. The resulting plasmid pPIC-HBscFv was linearized and transformed into P. pastoris GS115. The recombinant Pichia strains, identified by direct PCR and Zeocin-resistant screening of Pichia transformants, were cultured and induced with methanol. It was found that recombinant HBscFv, lead by alpha-factor, could be secreted into the culture supernatant to a level of 80mg/L. The bioactivity of Pichia produced HBscFv was confirmed by indirect ELISA, which also suggested that the bioactivity of HBscFv in the culture supernatant reached its peak in 72h and decreased in the late-stage of the induction. PAS staining suggests that HBscFv produced by yeast is poorly glycosylated or none-glycosylated protein.
Blotting, Western ; Electrophoresis, Polyacrylamide Gel ; Hepatitis B Surface Antigens ; immunology ; Humans ; Pichia ; genetics ; metabolism ; Polymerase Chain Reaction ; Single-Chain Antibodies ; genetics ; immunology ; metabolism

OBJECTIVERecent study demonstrated that hypoxia could regulate the proliferation and differentiation of neural stem cells in vitro. In the present study, effects of low glucose and/or hypoxia on the proliferation and metabolism of neural stem cells were investigated in vitro.

METHODSThe neural stem cells were isolated from the rat embryonic mesencephalon (E13.5), and exposed to different oxygen concentrations (low oxygen: 3% O2 or normoxia: 20% O2) and different glucose concentrations (high glucose concentration: 4.5 g/L and low glucose concentration: 1.4 g/L) for 3 days. The proliferation of neural stem cells were examined by CCK-8 assay. Furthermore, the content of glucose, lactate, and pyruvic acid in the medium were measured after cultured in different condition for 1, 3, 5 days.

RESULTSLow oxygen and low glucose could increase the proliferation of neural stem cells respectively; in addition, the number of neurospheres under both low oxygen and glucose was the most among the four groups. The content of glucose and pyruvic acid in the medium from low oxygen or low glucose condition decreased, while the lactate concentration increased compared with the control group.

CONCLUSIONThe results indicate the neural stem cells prefer grow under the low glucose and low oxygen condition, and that is mainly under going glycolysis to maintain its self-renew ability. This study may provide us a useful clue for application of neural stem cells transplantation.

Animals ; Cell Hypoxia ; Cell Proliferation ; Cells, Cultured ; Female ; Glucose ; metabolism ; Glycolysis ; Neural Stem Cells ; cytology ; metabolism ; Pregnancy ; Rats ; Rats, Sprague-Dawley

L_9(3~4) orthogonal experiment design was used to optimize the preparation of the patches,and investigate its affecting factors and skin irritation. Eugenol was taken as the index component to study the release behavior in vitro and percutaneous penetration of Cangai oil transfersomes patches by HPLC.The results showed that the optimal prescription for preparing Cangai oil transfersomes patches were Eudragit E100 0.6 g, succinic acid 0.08 g,triethyl citrate 0.25 g,glycerol 0.2 g.Patches prepared by the preferred preparation had a flat appearance without obvious bubbles.The initial adhesion was 18.33±2.52, the stickiness was(30.01±2.45) min,and the peel strength was(5.62±0.95) kN·m~(-1).The results of affecting factors experiment showed the order of factors affecting its adhesion was humidity>temperature>lighting,and the skin irritation test results showed no significant skin irritation after 24 h of single administration. The results of drug release behavior in vitro showed that the release and the percutaneous penetration of both Cangai oil patches and Cangai oil transfersomes patches conformed to the Higuchi equation.The release amount of eugenol were 80.66% and 82.25% at 72 h, with no significant difference. The cumulative permeation area of eugenol per unit area reached(0.195 6±0.065 9),(0.131 0±0.045 5) mg·cm~(-2) at 72 h, with significant differences(P<0.05).The experiment results proved that the preparation process of Cangai oil transfersomes patches was stable,and the prepared patches had a good adhesion. At the same time,the preparation of transfersomes patches could alleviate and control the release of the drug to a certain extent, and provide a certain experimental basis for clinical pediatric drug safety.
Administration, Cutaneous ; Drug Carriers ; Drug Liberation ; Humans ; Plant Oils/pharmacology* ; Polymethacrylic Acids ; Skin/drug effects* ; Skin Absorption ; Transdermal Patch

OBJECTIVETo investigate the clinico-pathologic features, treatment and prognosis of Castleman disease.

METHODSThe clinico-pathologic data of 16 patients diagnosed as Castleman disease from January 2002 to December 2014 were analyzed retrospectively.

RESULTSThe median age was 28.5 (7-73)years old. There were 14 unicentric cases, 92.8% (13/14) of which was diagnosed as hyaline-vascular type. Two multicentric cases was diagnosed as plasmatcyic type. All the patients were treated by surgical resection and their median follow-up was 55.5 (2-150)months. As a result, 13 unicentric cases achieved sustained remission, 1 unicentric case with plasmatocytic type relapsed at 60th month after surgical resection.

CONCLUSIONClinical subtype and histopathogenic type are the dominating progonostic factors in Castleman patients. The clinical presentation of unicentric disease has been found to be benigns and the surgical resection can be used as first-line treatment method in clinic. The clinical presentation of multicentric disease may be stable or advanced, and the prognosis of advanced cases is poor as there are no effective treatments.

Adolescent ; Adult ; Aged ; Castleman Disease ; Child ; Humans ; Middle Aged ; Prognosis ; Retrospective Studies ; Treatment Outcome ; Young Adult

This study aimed to provide data support for resource utilization of the stems and leaves of Astragalus membranaceus var. mongholicus(SLAM) by analyzing and evaluating the chemical constituents. The crude protein, crude fiber, and soluble saccharide of SLAM were analyzed by Kjeldahl method, filtration method, and UV-Vis spectrophotometry, respectively. The nucleosides, amino acids, flavonoids, and saponins of SLAM were analyzed by ultraperformance liquid chromatography-triple quadrupole mass spectrometry(UPLC-TQ-MS). Combined with principal component analysis(PCA), the quality difference of resource components of SLAM was comprehensively evaluated. The results showed that the average content of crude protein, crude fiber, total polysaccharide, and redu-cing sugar in SLAM was 5.11%, 30.33%, 11.03 mg·g~(-1), and 31.90 mg·g~(-1), respectively. Six nucleosides, 15 amino acids, 22 flavonoids, and one saponin were detected, with an average content of 1.49 mg·g~(-1), 6.00 mg·g~(-1), 1.86 mg·g~(-1), and 35.67 μg·g~(-1), respectively. The content of various types of chemical components in SLAM differed greatly in different harvesting periods and growing years. The results of PCA showed that the quality of SLAM produced in Ningxia was superior. The results can provide references for the utilization of SLAM.
Astragalus propinquus/chemistry* ; Gas Chromatography-Mass Spectrometry ; Flavonoids/analysis* ; Plant Leaves/chemistry* ; Amino Acids ; Saponins/analysis*