OBJECTIVE　To detect the trace glutaraldehyde in the fluid from venous circuit tube of hemodialysis,a HPLC assay was developed.METHODS　20 mL fluid taken from the venous circuit tube were derived with DNPH for 3 h,then filtered and injected. Chromatography was conducted on C18 column at 29℃.The mobile phase was consisted of 60% CH3N and 40% H3PO4 with the flow rate at 1 mL*min-1, and the detetion wavelength was at 365 nm.RESULTS　The concentrations of glutaraldehyde in the fluid ranged from 14.99 to 37.40 μg.mL-1.CONCLUSION　This HPLC method is simple and accurate to detect the trace glutaraldehyde remained in the fluid from venous circuit tube.

Objective To verify the indicator reference value ranges of blood cells analytic report by quoting the WS/T 405-2012 industry standard in Changsha area and to establish the reference value ranges of research parameters suitable for local area.Methods The blood cells analytic results in 3138 cases of reference group were analyzed according to the program of the Formulation of Reference Intervals in Clinical Laboratory Detection Items and the results were compared with the WS/T 405-2012 industry standard.Results WBC,Neu#,Mon%,MCHC,PLT,MPV,PCT,P-LCC,P-LCR and InR‰ in the report parameters and WBC-D,PDW-SD in the research parameters had no statistical differences between males and females(P>0.050);WBC,Neu#,Mon%,MCHC and PLT had no difference between genders,the indicators were narrowed compared with the reference value ranges in standard;the reference value ranges of Neu%,RBC,HGB,MCV and MCH in the male blood cells analytic indicators were narrowed,the lower limits of Bas%,Lym#,Mon# and HCT reference value ranges were elevated,the upper limits of Bas%,Lym#,Mon# and HCT reference value ranges were elevated,while the upper limits of Eos# and Eos% reference value ranges were decreased,the Lym% lower limit was decreased;Neu%,Lym#,Mon# and RBC in the female blood cells analytic indicators were narrowed,the lower limits of Bas# and Bas% were increased,the lower limits of Eos%,Lym%,HGB,MCV and MCH were decreased,the upper limits of Eos#,Eos%,Lym%,HGB,MCV and MCH were decreased and the HCT upper limit was elevated.Conclusion The Changsha area should have the reference value ranges of selective quoted standards and establishes the reference value ranges suitable for local area or laboratory.

Objective To study the efficacy and safety of four surgical techniques of tuberculosis of lumbosacral junction retrospectively. Methods Between Jul 2001 and Jan 2013, 79 patients with lumbosacral spinal tuberculosis underwent surgery. Antituberculous chemotherapy and nutrition support prior to surgery were used for at least two weeks. 45 patients underwent single stage radical debridement, fusion and anterior instrumentation (A group). 18 patients underwent combined anterior and posterior spinal surgery (AP group), 10 patients underwent transpedicular drainage, posterior instrumentation, and fusion (P group), and 6 patients underwent anterior radical debridement (D group). All the patients were treated by antituberculous chemotherapy for 18 months and followed regularly. The operation duration, blood loss, clinical status, ESR, VAS, ODI, roentgenogram and 3D?CT were concerned to estimate the progress of tuberculosis. Radiographs were analyzed before surgery, immediately after surgery, and at the final follow?up examination to assess the result of anterior fusion and maintenance of correction. Results There was no inju?ry of blood vessel, ureter or cauda equina during surgery. The mean follow?up period was 23 months (range 18-42 months). No obvious loss of deformity correction was observed. There was no recurrence, no tuberculous peritonitis, and no incidence of im?potence or retrograde ejaculation in any of these patients. The average operating duration(min) were 144.31 ± 23.18, 444.72 ± 141.63, 351.50 ± 85.25, 90.00 ± 29.66, respectively; The average blood loss(ml)were 266.67 ± 104.45, 988.99 ± 488.26, 890.00 ± 306.23, 200.00±104.88, respectively; The average Pre?op VAS were 4.71±1.79, 5.22±1.48, 3.30±1.64, 2.50±1.52, respectively;The average last follow?up VAS were 0.89±0.68, 0.90±0.74, 1.00±0.63, respectively; The average Pre?op ODI(%)were 29.64± 7.85, 32.17±7.59, 28.20±4.26, 20.67±4.63, respectively; The average last follow?up ODI(%)were 5.09±3.59, 4.78±3.78, 4.80± 3.39, 4.00 ± 1.18, respectively; The average Pre?op lumbosacral angle(°)were 20.61 ± 4.92, 23.78 ± 5.84, 25.10 ± 4.28, 21.67 ± 4.27, respectively; The average Post?op lumbosacral angle were 27.17±3.66, 30.56±5.31, 32.10±4.01, 24.83±2.32, respectively;The average last follow?up lumbosacral angle were 23.89 ± 3.12, 27.00 ± 5.46, 29.00 ± 4.85, 23.33 ± 2.50, respectively. Conclu?sion Single stage anterior interbody fusion with anterior instrumentation worked effectively to stabilize lumbosacral junction (less invasive, short surgical duration, no injury of posterior column). Anterior interbody fusion combined with posterior instrumentation was recommended for patients with extensive bone defect and low iliocava junction.

Objective To analyse the phenotypes of the drug?resistant tuberculosis, and investigate the outcomes of the individualize surgery and chemotherapy for these patients. Methods From January 2009 to June 2012, we retrospectively ana?lyzed 49 patients with drug?resistant tuberculosis spondylitis admitted in Southwest Hospital. 33 were initial cases and 16 were re?curring cases. All the 49 patients received individualized open operation or CT?guided percutaneous drainage and local chemother?apy depending on the characteristics of the focus. Individualized chemotherapy regimens were tailored for all patients according to the drug?resistant spectrum and all patients were followed up successfully at least 24 months. All the clinical data were collected and analyzed by statistical methods. Results Among the 49 patients, 14 were monoresistance tuberculosis, 11 were polyresis?tance tuberculosis, and 24 cases were multi?drug resistant tuberculosis. Frequence of the drug?restistance from high to low was Iso?niazid, Rifampicin, Streptomycin, Levofloxacin, Dipasic/Rifapentine, Ethambutol, Protionamide, Capreomycin, Paza?aminosalicy?late, and Amikacin. 43 patients received open operation and 6 patients received CT?guided percutaneous drainage and local che?motherapy. Time of the percutaneous drainage was (48±11) days (39-60 days), and all patients received Individualized chemother?apy with an average of (29.5±2.5) months (24-36 months) postoperatively. At the last follow?up, all patients had remarkable pain remission, 44 patients with paraplegia got slight or remarkable recovery and 17 patients with kyphosis got significant correction. Conclusion The main drug?resistant spectrums are Isoniazid、Rifampicin、Streptomycin、Levofloxacin. The individualized sur?gery combined with individualized chemotherapy made according to the drug?resistance is a feasible treatment for the drug?resis?tant tuberculosis especially the multi?drug resistant tuberculosis.

OBJECTIVETo construct a universal, highly attenuated orf virus expression vector for exogenous genes using green fluorescent protein (GFP) as the reporter gene.

METHODSThe flanking regions of the ORFV132 of orf virus DNA were amplified by PCR to construct the shuttle plasmid pSPV-132LF-EGFP-132RF. The shuttle plasmid was transfected into OFTu cells and GFP was incorporated into orf virus IA82Delta 121 by homologous recombination. The recombinant IA82Delta121-V was selected by green fluorescent signal. The deletion gene was identified by PCR and sequencing. The effects of ORFV132 knockout were evaluated by virus titration and by observing the proliferation of the infected vascular endothelial cells in vitro.

RESULTSThe recombinant orf virus IA82Delta121-V was obtained successfully and quickly, and the deletion of ORFV132 did not affect the replication of the virus in vitro but reduced its virulence.

CONCLUSIONGreen fluorescent protein is a selectable marker for rapid, convenient and stable selection of the recombinant viruses. Highly attenuated recombinant orf virus IA82Delta121-V can serve as a new expression vector for exogenous genes.

Cells, Cultured ; Endothelial Cells ; metabolism ; Gene Deletion ; Genes, Reporter ; Genetic Vectors ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Humans ; Orf virus ; Plasmids ; Transfection

Objective To investigate the treatment outcome,laryngeal preservation and side-effect in locally advanced hypopharyngeal carcinoma treated with combined Hilical tomotherapy (HT) or intensitymodulated radiotherapy (IMRT) and chemotherapy and/or EGFR inhibitor (Cetuximab or Nimotuzumab).Methods A total of 68 patients (20 cases with T1-2N1-3M0 and 48 cases with T3-4N1-3M0) with locally advanced hypopharyngeal cancer were treated individualy with non-surgical combined modality treatments including induction chemotherapy followed by concurrent chemoradiotherapy,induction chemotherapy followed by concurrent radiotherapy and EGFR inhibitor,concurrent chemoradiotherapy and EGFR inhibitor,and concurrent radiotherapy and EGFR inhibitor.HT was used in 40 patients and IMRT in 28 patients.Side-effects were evaluated with the established Common Terminology Criteria for Adverse Events (CTCAE) version 3.0 criteria.Results The average follow-up time was 25.7 months (range 3-69 months).All patients completed the planned radiotherapy without treatment breaks,and 66 (97.0％) of 68 patients completed the planned chemotherapy.The 2-year and 3-year overall survival rates were 78.8％ and 64.7％ respectively,with an organ preservation rate of 84.2％.The most common side-effect greater than or equal to grade 3 was oropharyngeal mucositis.No patient dependent on a percutaneous gastrostomy and tracheostomy tube.Conclusion Hypopharyngeal carcinoma can be treated with non-surgical combined modality treatment including HT or IMRT,with a high laryngeal organ preservation rate and minimal toxicities.

ObjectiveTo investigate the effects of maltol aluminum exposure on miR-193a-3p, demethylase AlkB homolog 5 (ALKBH5), phosphatase and tensin homolog deleted on chromosome ten (PTEN) and protein kinase B (AKT), and whether miR-193a-3p is involved in aluminum-induced cognitive impairment by regulating ALKBH5/PTEN/AKT signaling pathway. Methods Specific pathogen-free male SD rats were randomly divided into control group and low-, medium- and high- dose groups according to their body weight, with eight rats in each group. Rats in the low-, medium-, and high- dose groups were intraperitoneally injected with maltol aluminum solution at concentrations of 10.00, 20.00, and 40.00 μmol/kg body weight, respectively, while the rats in control group were given an equal volume of 0.9% sodium chloride solution. Rats were injected for five days every week for three months. After injection, the novel object recognized test was used to assess the learning and memory ability of the rats. The relative expression of miR-193a-3p and B-cell lymphocytoma-2 (Bcl-2), Bcl-2 associated X protein (Bax) and cysteine aspartate protease-3 (Caspase-3) mRNA in rat hippocampus was detected using the real-time quantitative polymerase chain reaction. The relative protein expression of ALKBH5, PTEN, and AKT2 in the rat hippocampus was detected using Western blot. Results The discrimination index and the preference index of the new object recognition test of the rats in high-dose group were lower than those in control group and low-dose group (all P<0.05). The relative expression of miR-193a-3p and Bcl-2 mRNA in the hippocampus of the rats in high-dose group was lower than those in control group and low-dose group (all P<0.05). The relative mRNA expression of Bax in the high-dose group was higher than those in the control group and low-dose group (both P<0.05). The relative mRNA expression of Caspase-3 of the rats in the high-dose group was higher than that in the other three groups (both P<0.05). The relative protein expression of ALKBH5 in the hippocampus of the rats in the high-dose group was lower than that in the control group (P<0.05). The relative expression of PTEN protein was higher than those in the control group and low-dose group (both P<0.05). The relative protein expression of AKT2 was lower than those in the control group and low-dose group (both P<0.05). Conclusion Sub-chronic aluminum exposure can inhibit the expression of miR-193a-3p in the hippocampus of rats, which may disrupt the ALKBH5/PTEN/AKT pathway and affect normal neuronal homeostasis and cellular function. This pathway may play an important role in aluminum-induced cognitive impairment.

Objective: To explore the relationship between adolescents physical fitness and depressive symptoms, and to provide reference for the early prevention and intervention of depressive symptoms and improvement of physical fitness in Chinese adolescents. Methods: From September to December 2021, a total of 8 102 adolescents were selected by random cluster sampling method in Shanghai, Urumqi, Changsha and Kunming. The Center for Epidemiologic Studies Depression Scale(CES-D) was used to investigate the depressive symptoms, and completed grip strength, standing long jump, 50 m running, modified sitting forward flexion, 20 s repeated traverse, 30 s sit ups, 20 m round trip running (20 m SRT) test. χ 2 test, Goodman Kruskal Gamma and Logistic regression analysis were used to analyze the relationship between physical fitness index(PFI) and depressive symptoms. Results: The overall detection rate of depressive symptoms in adolescents with high level PFI was 23.4%, and the detection rate of low level adolescents was 26.3%, with a statistically significant difference ( χ 2=6.73, P =0.01). There was a significant positive correlation between PFI and depressive symptoms in the high school group ( G=0.09, P <0.05) and the boy group ( G=0.12, P < 0.05 ), and there was no significant association between PFI and depressive symptoms in the junior high school group and the girl group ( P >0.05). After adjusting for gender and age in the Logistic regression model, compared with those with high PFI, the risk of depressive symptoms in those with low PFI was 1.18 times (95% CI =1.05-1.33). Conclusion There is a correlation between physical fitness and depressive symptoms in adolescents. Adolescents with low PFI are at higher risk of developing depressive symptoms than those with high PFI.

OBJECTIVE: To carry out genetic and metabolite analysis for an infant with cerebral creatine deficiency syndrome type 2 (CCDS2). METHODS: Clinical data of the child was collected. Whole-exome sequencing was carried out to identify potential variants by next generation sequencing. Candidate variants were confirmed by Sanger sequencing. Metabolites were determined by tandem mass spectrometry and magnetic resonance spectroscopy. Treatment was carried out following the diagnosis and genetic counseling for the affected family. RESULTS: Two novel heterozygous variants (c.289delC and c.392-1G>C) of the GAMT gene were identified in the proband, which were respectively inherited from her father and mother. In silico analysis suggested both variants to be pathogenic. Creatine (Cr) level of the child was very low, and cerebral guanidinoacetate (GAA) level was slightly increased. But both had recovered to normal in two weeks, and cerebral Cr level was significantly improved after two months. Intellectual and motor development of the child were significantly improved. CONCLUSION The child was diagnosed with CCDS type 2, for which pathogenic variants of the GAMT gene may be accountable. Treatment has attained a satisfactory effect for the patient.

Objective:To explore the relationship between pathogens in the olfactory cleft area and olfactory disorders in patients with upper respiratory inflammation (URI) during the prevention and control of 2019 novel coronavirus disease (COVID-19).Methods:A total of 234 URI patients including acute upper respiratory infection, chronic rhinosinusitis (CRS), allergic rhinitis (AR) were continuously selected from September 2020 to March 2021 in Beijing Anzhen Hospital and 98 healthy adults were enrolled as controls. The secretions from the olfactory cleft of all subjects were collected with nasal swabs under nasal endoscopy. Multiple real-time fluorescent quantitative polymerase chain reaction detection method was used to detect nucleic acids of 33 types of respiratory pathogenic microorganism. Sniffin′ Sticks olfactory test was performed on all patients with URI. URI patients with olfactory dysfunction were followed up for 9 (8, 10) months ( M ( Q1, Q3)). SPSS 20.0 software was used for statistical analysis. Results:Among the 98 controls, 9 (9.18%) were positive for pathogenic microorganisms, including 1 (1.02%) rhinovirus, 1 (1.02%) parainfluenza virus type 3, 3 (3.06%) enterovirus, 1 (1.02%) staphylococcus aureus and 3 (3.06%) Moraxella catarrhalis. Among the 234 URI patients, 111 (47.44%) had olfactory disorders and 123 (52.56%) had normal sense of smell. In the olfactory disorder group (111 cases), 38 cases (34.23%) were positive for pathogenic microorganisms, and 4 cases (3.60%) were mixed infection, including 11 cases of rhinovirus (9.91%), 5 cases of coronavirus 229E (4.50%), 2 cases of coronavirus OC43/NL63 (1.80%), 3 cases of parainfluenza virus type 1 (2.70%), 2 cases of enterovirus (1.80%), 1 case of influenza B virus type BV (0.90%), 11 cases of Staphylococcus aureus (9.91%), 7 cases of Moraxella catarrhalis (6.31%), and 1 case of Klebsiella pneumoniae (0.90%). In the normal smell group (123 cases), 18 cases (14.63%) were positive for pathogenic microorganisms, and 1 case (0.81%) was mixed infection, including 3 cases of rhinovirus (2.44%), 4 cases of coronavirus 229E (3.25%), 1 case of Influenza virus type 3 (0.81%), 3 cases of enterovirus (2.44%), 3 cases of Staphylococcus aureus (2.44%), 4 cases of Moraxella catarrhalis (3.25%), and 1 case of Klebsiella pneumoniae (0.81%). Univariate analysis between the two groups found that there were significant differences in the detection rate of pathogenic microorganisms, rhinovirus and Staphylococcus aureus between the groups (all P<0.05). The detection rate of parainfluenza virus type 1, Staphylococcus aureus, and rhinovirus were different between the patients with olfactory disorder and normal olfactory function in the three subgroups of acute upper respiratory tract infection, CRS and AR, respectively (χ 2 value was 3.88, 4.53 and 4.73, respectively, all P<0.05). During the follow-up period, among the 111 patients with olfactory disorder, 71 (63.96%) patients′ olfactory function returned to normal, 32 (28.83%) patients′ olfactory function improved but not completely returned to normal, 8 (7.21%) patients′ olfactory function did not improve. Conclusions:During the prevention and control of COVID-19, rhinovirus or Staphylococcus aureus infection or colonization of URI patients is closely related to olfactory disorders. Parainfluenza virus type 1 infection can cause relatively persistent olfactory disorders in patients with acute upper respiratory tract infection. Staphylococcus aureus and rhinovirus colonization are related to the occurrence of olfactory dysfunction in CRS and AR patients respectively.