Objective To evaluate the characteristics of dose distribution of neuronal networks in vitro on microelectrode arrays(MEAs)under 2.6 GHz radiofrequency(RF)exposure.Methods The MEAs were coupled with a real-time RF exposure setup,and electromagnetic simulation software was used to calculate the RF dose absorbed in cultured neuronal networks.A fiber-optic temperature probe was used for experimental validation and monitoring of the cell temperature during RF exposure.The MEAs were used to record the electrical activity of neurons.Results For an input power of 1 W,a specific absorption rate(SAR)level of(15.51±2.48)W/kg was calculated,and the variability of the SAR distribution was 16%.In our experimental system,the temperature elevation of neurons was up to 0.15℃for an SAR of 4 W/kg RF exposure.Conclusion The exposure device can provide high SAR efficiency and uniformity in the 2.6 GHz band,which is suitable for studying the real-time effects of RF fields on the electrical activity of neuronal networks in the 5G network band.

Objective: To investigate the distribution of toxoplasma cysts in the brain of infected mice and the effect of pathological changes on the behavior and neuropsychiatry of the mice during chronic infection with Toxoplasma gondii( T．gondii) ． Methods : Mice were infected with Prugniaud strain of T．gondii by oral gavage．The brain tissues of infected mice were collected on the days of 10，30，40，90，120 and 160 after infection respectively，and the hippocampal hypothalamus，prefrontal lobe ，striatum and cerebellum regions were separated．The number of cysts and neuropathological changes in each infected area were observed and recorded by HE staining．The number of cysts and neuropathological changes in each infected area were observed and recorded. Results : T．gondii infected mice showed symptoms of vertical hair and arched back，which were the most significant on the 40th day，and then gradually recovered with hemiplegia and circling in circles． At each time point ，the number of toxoplasma cysts was the largest in hippocampal hypothalamus，followed by prefrontal lobe and striatum，and the least in cerebellum．The diameter of toxoplasma cysts increased with time．During chronic infection，specific pathological manifestations of toxoplasma encephalitis，such as neuronophagy，were observed in all regions of the brain tissue．The above pathological changes of toxoplasma encephalitis reached the peak on the 40th day，and gradually recovered， and increased to the stimulation peak on the 120th day，and then gradually recovered． Conclusion The behavioral and neuropsychiatric symptoms of T．gondii during chronic infection were correlated with the localization and distribution of toxoplasma cysts in the brain of infected mice，and showed dynamic changes.

Objective:To compare 3D-printing-assisted surgery and conventional surgery in the treatment of Schazker type Ⅵ tibial plateau fractures.Methods:A retrospective study was conducted to analyze the clinical data of 50 patients with type Ⅵ tibial plateau fracture who had been treated from January 2019 to December 2021 at the 5 Departments of Orthopedics in The First Affiliated Hospital of Nanchang University, The First People's Hospital of Jiujiang, Pingkuang General Hospital, Ganzhou People's Hospital, and Nanchang Hongdu Hospital of Traditional Chinese Medicine. The patients were divided into 2 groups according to their different treatment methods. In the 3D printing group of 25 cases treated by 3D-printing-assisted surgery, there were 14 males and 11 females, with an age of (42.5±9.1) years; in the conventional group of 25 cases treated by conventional surgery, there were 13 males and 12 females with an age of (42.2±9.3) years. The 2 groups were compared in terms of operation time, intraoperative blood loss, intraoperative fluoroscopy frequency, fracture healing time, postoperative complications, the Rasmussen radiological scores and the American Hospital for Special Surgery (HSS) knee function scores at 6 and 12 months after operation.Results:There was no significant difference in the preoperative general data between the 2 groups, indicating comparability ( P>0.05). The operation time [(125.4±10.6) min], intraoperative blood loss [(206.2±16.3) mL], intraoperative fluoroscopy frequency [(9.2±2.7) times] and fracture healing time [(3.0±0.7) months] in the 3D printing group were all significantly less than those in the conventional group [(168.2±14.1) min, (303.2±20.4) mL, (15.5±3.5) times and (4.1±0.8) months] while the Rasmussen radiological scores (17.6±1.2 and 17.9±0.6) and HSS knee scores (90.8±6.4 and 91.5±5.6) at 6 and 12 months after operation in the 3D printing group were all significantly higher than those in the conventional group (16.2±2.6 and 16.7±2.2; 84.5±9.2 and 87.6±8.0) (all P<0.05). In the 3D printing group, there were 1 case of wound infection and 1 case of wound dehiscence after operation. In the conventional group, there were 2 cases of wound skin necrosis, 3 cases of wound dehiscence, 1 case of traumatic arthritis, 2 cases of wound infection, and 1 case of screw loosening. The incidence of complications in the 3D printing group (8.0%, 2/28) was significantly lower than that in the conventional group (36.0%, 9/25) ( P<0.05). Conclusion:In the treatment of Schatzker type VI tibial plateau fractures, compared with conventional surgery, 3D-printing-assisted surgery can lead to better curative outcomes, because it is conducive to lowering surgical difficulty, reducing postoperative complications, and promoting fracture union and functional recovery of the knee.

Objective:To identify the genetic variation in a mucopolysaccharidosis type Ⅱ(MPS Ⅱ)family, and conduct a functional study of iduronate-2-sulfatase(IDS): c.323A>C.Methods:A five-generation MPS Ⅱ family of 83 individuals including 4 patients from northern China was collected. Urine mucopolysaccharide and Alder-Reilly body were tested to assist the clinical diagnosis of MPS Ⅱ. IDS enzyme activity was detected on core family members. By the whole exome sequencing of a MPS Ⅱ patient in this family and bioinformatics analysis, the variant was screened and further identified by PCR-Sanger sequencing. Finally, to validate the function of the variant in vitro, the wild-type IDS overexpression plasmid(pCMV-hIDS-WT)and the IDS overexpression plasmid carrying the mutation site(pCMV-hIDS-c.323A>C)were transfected into COS-7 cells and the IDS activity was detected. Results:The proband(Ⅳ3)and Ⅳ4 were diagnosed as MPS Ⅱ by urine mucopolysaccharide, Alder-Reilly body, and IDS enzyme activity tests. Ⅳ3, Ⅳ4, Ⅲ19, and Ⅲ32 were determined to carry IDS: c.323A>C missense variant through the whole-exome sequencing, and diagnosed as MPS Ⅱ. Meanwhile, Ⅱ2, Ⅱ4, Ⅱ8, Ⅱ12, Ⅱ14, Ⅲ5, Ⅲ7, Ⅳ14 in the MPS Ⅱ family carried IDS: c.323A>C missense variant, and were excluded as MPS Ⅱ. The in vitro experiment in COS-7 cells showed that the missense mutation led to a significant decrease in IDS enzyme activity. Conclusion:The variant IDS: c.323A>C: p.Y108S significantly decreases the activity of IDS enzyme in vivo and in vitro, and it is identified as a pathogenic variant for MPS Ⅱ.

Objective:To evaluate the effects of CD34 + cell transplantation on radiation-induced brain injury (RIBI) and the relationship with the activity of astrocytes in rats. Methods:Healthy adult male Sprague-Dawley rats, weighing 210-230 g, were divided into 3 groups ( n=36 each) using a random number table method: control group (C group), RIBI group, and CD34 + group.RIBI model was established by computed tomography (CT) scanning in anesthetized rats.Another 6 rats were selected, and CD34 + cells were eluted by flow cytometry and labeled with BrdU.CD34 + cells were transplanted at day 7 after establishing the model.Brain tissues were obtained at 7, 14 and 28 days after establishing the model in C and RIBI groups and at 14 and 28 days after establishing the model in CD34 + group for determination of Evans blue (EB) extravasation ratio and expression of GFAP (by immuno-histochemistry). Results:Compared with group C, the EB extravasation ratio was significantly increased after establishing the model, and the expression of GFAP was up-regulated in group RIBI ( P<0.05), and no significant change was found in EB extravasation ratio after establishing the model in group CD34 + ( P>0.05). Compared with group RIBI, the EB extravasation ratio was significantly decreased after establishing the model, and the expression of GFAP was down-regulated in group CD34 + ( P<0.05). Conclusion:CD34 + cell transplantation can reduce RIBI, and the mechanism may be related to inhibiting the activity of astrocytes in rats.

OBJECTIVE：  To improve the method for the content determination of astragaloside Ⅳ in Xiangju granules， and to evaluate the consistency of relevant preparations with the components of original formulation， so as to provide evidence for the modern preparation of TCM compound. METHODS： HPLC-ELSD method was established for the content determination of astragaloside Ⅳ in Xiangju granules， and compared with original standard TLC scanning. Using critrinin， ferulic acid， calycosin glucoside， liquiritin， glycyrrhizic acid， rosmarinic acid， buddleoside and magnoline as control， HPLC method was used to determine the release components of self-made Xiangju granules， Xiangju capsules， Xiangju tablets in water. Fingerprint characteristics chromatogram of different Xiangju preparations and original formulation extract were compared by using Similarity Evaluation System for Chromatographic Fingerprint of TCM （2012 version）. At the same time， HPLC-ELSD method was used to determine and compare the release rate of astragaloside Ⅳ from different Xiangju preparations and original formulation extract in water. RESULTS： Established HPLC-ELSD method was specific. The linear range of astragaloside Ⅳ was 0.13-2.10 mg/mL. RSDs of precision， repeatability and stability tests were all lower than 3％ （n＝6）， and average recovery was 97.66％ （RSD＝1.01％，n＝6）. Average content of astragaloside Ⅳ by this method was 0.398 mg/g （RSD＝1.01％， n＝3）， which had better reproducibility than TLC scanning. The comparative results of characteristic fingerprints showed that the similarity among Xiangju granules， Xiangju capsules， Xiangju tablets and the original formulation dry extract powder was more than 0.850. Average release rates of astragaloside Ⅳ in Xiangju granules， Xiangju capsules， Xiangju tablets and the original formulation extract were 0.392， 0.358， 0.349， 0.389 mg， respectively. Compared with original formulation extract， there was no statistical significance in release rate of astragaloside Ⅳ in Xiangju granules （P＞0.05）， while there was statistical significance in Xiangju capsules and Xiangju tablets （P＜0.01）. CONCLU- SIONS： Established HPLC-ELSD method is accurate and feasible， and is suitable for the content determination of astragaloside Ⅳ in Xiangju granules. The main components of Xiangju granules are consistent with original formulation.

Objective To investigate the immunodiagnostic valueofrecombinant granulocvte protein 5(rGRA5) protein in prokaryotic Toxoplasma gondii.Methods The PCR was used to ampliify GRA5 gene,then insert the target gene into the prokaryotic expression vector pET28a,and identified by double digestion and sequencing, then transfecte the correct targert gene into BL21 competent cells, SDS-PAGE and Western blot were used to detect the expression of the recombinant protein in BL21.ELISA was used to detect the serum of 100 cases of serologically Toxoplasma-positive individuals.Results The products of GRA5 gene of 363 bp in length was successfully amplified by PCR,the prokaryotic expression vector pET28a-GRA5 was constructed successfully, the result of double digestion showed that the insert fragment size was correct, and DNA sequencing results showed that the homology of GRA5 gene with GenBank was 100%.Also the expression of GRA5 protein was successfully detected in BL21 by Western blot(about 14 ku).ELISA method was used to detect 100 cases of patients with 73 cases showed positive results, the positive diagnosis rate was 73.0%.Among them, the positive detection rate of IgG positive samples was 72.5% in 40 cases,the positive detection rate of IgM positive samples was 53.3% in 30 cases,the positive detection rate of IgG and IgM positive samples was 93.3% in 30 cases.Conclusion The prokaryotic expression vector pET28a-GRA5 is constructed successfully, and the recombinant protein has potential for immunological diagnosis of toxoplasmosis.

A rapid and sensitive method of loop-mediated isothermal amplification(LAMP) was established to detect Pseudomonas aeruginosa(P.aeruginosa).Three pairs of LAMP primers(inner,outer and ring primers) were designed according to the gbca gene of P.aeruginosa.Since adding hydroxy naphthol blue(HNB) to the reaction system, a positive reaction was indicated by a colorchange before and after the reaction,and was verified by agarose gellectrophoresis.Both LAMP and PCR were applied to detect clinical specimens, the sensitivity and specificity of the detection method were evaluated,and were compared with those of conventional PCR.A LAMP method for detecting P.aeruginosa was successfully established.The LAMP method showed specificity for P.aeruginosa without other bacteria amplification.The established LAMP method in this study enables rapid,sensitive and specific detection of P.aeruginosa,and can be applied for grass roots and small scale laboratories as well as field surveillance.

Objective: To investigate the comparative study of serum hepatitis B virus (HBV) large protein (HBV-LP) , HBV-DNA, and Pre S1 antigen (Pre S1-Ag) detection in the evaluation of serum HBV replication in patients with chronic hepatitis B. Methods: A total of 482 patients infected with chronic hepatitis B virus (CHB) were enrolled and the serums were collected in a hospital of Hefei city in Anhui province from June 2013 to March 2015. The serum HBV-LP, HBV markers(HBV-M) and Pre S1-Ag were detected using ELISA, and HBV-DNA were quantified using quantitative real-time PCR. The positive detection rate difference of HBV-DNA, HBV-LP and Pre S1-Ag were compared, the correlation between the logarithm of HBV-DNA copies number and the absorbance value of HBV-LP was analyzed using Spearman rank correlation. Results: The positive rates of HBV DNA, HBV-LP, and Pre S1-Ag were 67.22% (324/482), 73.86% (356/482), and 37.34% (180/482), respectively (P<0.01). The positive rates of the three markers were 54.57% (185/339), 64.90% (220/339), and 27.73% (94/339), respectively, in 339 HBeAg-negative CHB patients (P<0.01). In HBeAg negative patients, the positive rate of HBV-LP in 185 cases of HBV-DNA positive samples was 90.81% (168/185), which was higher than that of Pre S1-Ag with rate of 39.46% (73/185) (P<0.01).The positive rate of HBV-LP was 33.77% (52/154) in 154 cases of patients with negative HBV-DNA whose positive rate was higher than Pre S1-Ag with positive rate of 13.64% (21/154)(P<0.01). The positive rates of HBV-DNA, HBV-LP and Pre S1-Ag in HBsAg, HBeAg and HBcAb positive groups were 97.04% (131/135), 94.81% (128/135), and 60.00% (81/135), (P<0.01); The positive rates of three indexes of HBsAg, HBeAb, HBcAb positive group were 53.74% (122/227), 63.88% (145/227), and 27.31% (62/227); The positive rates of three indexes of HBsAg and HBcAb positive group were 55.79% (53/95), 67.37% (64/95), and 28.42% (27/95) (P<0.01). The absorbance value of HBV-LP was positively related with the logarithm of HBV-DNA copies number (Spearman rank correlation coefficient was 0.908, P<0.01). Conclusion There was a good correlation between HBV-LP and HBV-DNA load value, and could be used as an effective complement for the detection of HBV-DNA and HBV-M. Compared with Pre S1-Ag.

Objective To research the immuno-protection of SJIR-2 DNA vaccine with nanometer microspheres a-gainst Schistosoma japonicum infection in mice. Methods To construct eukaryotic expression plasmid pEGFP-SJIR-2, identified by double digestion and sequenced delivery. The recombinant plasmid pEGFP-SJIR-2 was ex-tracted and was encapsulated into PLGA nanometer microspheres which were modified by CHS. 40 female BALB/c mice were randomly divided into 4 groups (n=10), each group of mice were injected with PBS, empty pEGFP plasmid, CHS-PLGA nanometer microspheres and CHS-PLGA-pEGFP-SJIR-2 nanometer microspheres 100 μg, re-spectively. Two weeks after the last immunization, each mouse was infected by cercaria of Schistosoma japonicum, sera of mice in each group were collected before each immunization and challenge infection. ELISA was used to de-tect the change of IgG in each group of micesera. 42 days later, all mice were sacrificed. The adult worms and eggs were collected and counted, and the worm and egg reduction rates were calculated as well. Results The recombi-nant plasmid pEGFP-SJIR-2 was successfully constucted, and there was significant difference in the numbers of worm and egg between CHS-PLGA-pEGFP-SJIR-2 group and PBS group ( P<0. 01 ) . The worm andegg reduction rates in CHS-PLGA-pEGFP-SJIR-2 group were 37. 36% and 46. 82% respectively. The IgG levels in mice sera of CHS-PLGA-pEGFP-SJIR-2 group were remarkably higher (P<0. 01) compared with PBS group. On the contrary, there was no significant difference between both pEGFP plasmid group and CHS-PLGA group in the numbers of worm and egg compared with PBS group. Conclusion SJIR-2 nanometer microspheres nucleic acid vaccine has some immuno-protection against Schistosoma japonicum infection in BALB/c mice,while it is worth further studying for it’ s potential value to be a candidate antigen molecule of Schistosoma japonicum vaccine.