Chromatography, High Pressure Liquid ; methods ; Drug Residues ; analysis ; Electrochemistry ; Ethidium ; analysis ; Luminescent Measurements ; methods ; Sensitivity and Specificity

Objective To analyse the clinical characteristics,gene mutation and enzymatic activity of αgalactosidase A(α-GalA)in a 15-year-old male patient with typical Fabry disease,whose mother was without any clinical manifestations.Methods Clinical features and laboratory data were collected from the patient and his mother.Genomie DNA was extracted from peripheral blood of the patient.his mother,and a healthy control subject.Seven exons of the GLA gene were amplified by PCR.PCR products were purified.cloned into T vector,and then sequenced.The enzymatic activity of α-GalA Was measured by fluorimetrie substrate assay. Results DNA sequencing results showed that a missense mutation of 10036-10038delAAG in exon 7 WaS identified in the patient,resulting in the replacement of 374 lysine and 375 glyeine by arginine,which Was not previously reported.The patient Was a hemizygote with gene mutation,his mother WaS a heterozygote carrying gene mutation,and the healthy control without mutation.α-GalA enzymatic activity assay showed that the enzymatic activity of the patient with GLA gene mutation was only 50%of the healthy control subject,while the enzymatic activity of the patient's mother Was about 70%of the heahhy control SObject.Conclusiolls Detecting GLA gene mutation and α-GalA enzymatic activity in patients with Fabry disease who have been clinically diagnosed seelns to be helpful in finding other patients in the family and in further understanding the molecular pathogenesis of that disease.

Objective To review the experience of colon interposition in the treatment of benign esophageal stricture.Methods 58 patients who had undergone colon interposition for esophageal replacements were studied retrospectively,including 53 patients with corrosive burn esophageal strictures,3 traumatic esophageal strictures and 2 congenital esophageal strictures.The interposition colon for all patients went through substemum paths.Results There was no postoperative death in the duration of hospital stay.14 cases developed postoperative complication including 2 total colon necrosis,7 anastomotic leak,2 anastomotic stricture and 3 recurrent laryngeal nerve injury.52 patients were followed-up(ranged 1 to 16 years),40 cases were extremely satisfied(1 grade),9 very satisfied(2 grade),2 satisfied(3 grade)and 1 unsatisfied(4 grade).Conclusion Colon interposition is an ideal procedure for esophageal benign stricture.

PurposeAs a main method to detect thyroid nodules, ultrasonography seems to have a rather low accuracy in differentiating benign and malignant ones. The present study aims to explore the diagnostic value of thyroid imaging reporting and data system (TI-RADS) in identifying benign and malignant thyroid nodules.Materials and Methods A total of 168 patients with thyroid nodules conifrmed pathologically (with 251 suspicious nodules) underwent ultrasonography and were further grouped into category 3-5 according to TI-RADS classiifcation standard. The results were retrospectively compared with histopathological ifndings. The sensitivity, speciifcity, accuracy, positive predictive value and negative predictive value of TI-RADS classiifcation in the diagnosis of benign and malignant thyroid nodules were calculated, and the differences in ultrasonic features between benign and malignant thyroid nodules were also compared.Results The surgical and pathological findings showed that 94 nodules were benign and 157 were sinister; TI-RADS regarded 93 nodules were benign and 185 were malignant. The difference of the two means of identification was significant (χ2=149.6,P<0.01). The sensitivity, speciifcity, accuracy, positive predictive value and negative predictive value of TI-RADS were 91.2% (144/157), 85.1% (80/94), 89.3% (224/251), 91.2% (144/158) and 86.0% (80/93), respectively. The ultrasonic manifestations of benign and malignant nodules were signiifcantly different with regard to aspect ratio, echogenicity, shape, calciifcation inside the nodule (χ2=8.7-121.4,P<0.01).Conclusion TI-RADS classiifcation standards have a high accuracy in the differentiation of benign and malignant thyroid nodules as a means of ultrasound examination, and thus can serve as an important guiding method in clinical diagnosis and treatment.

Objective To prepare a targeted antitumor drug delivery system using large-inner-diameter multi-walled carbon nanotubes (LID-MWCNTs) for sustained release and to study its performance. Methods LID-MWCNTs were puri?fied and oxidized,then use nanocarriers and USTs as homologous blockers. Folic acid and fluorescent labels were conjugat?ed onto the external surfaces of nanocarriers. CDDP (cisplatin) was encapsulated and ultrashort tubes (USTs) were employed to block the drug entry/exit paths. The microstructure of resulted drug delivery system (DDS) was observed, while drug load?ing efficiency and drug release profile in vitro were determined. The tumor-targeting property and cytotoxicity of DDS were also assessed. Results LID-MWCNT based sustained release targeted drug delivery system was established. Drug loading efficiency of CDDP@UST-FA-LID-MWCNTs was as high as 70.97%. A typical biphasic sustained release pattern was dem?onstrated, and the accumulating release time was 18 h. DDS exhibited a certain kind of tumor-targeting property, and inhibit?ed proliferation of tumor cells in a dose-dependent manner. Conclusion CDDP@UST-FA-LID-MWCNT drug delivery system exhibited an improved drug loading efficiency and a sustained drug release profile. It could specifically target the tu?mor cells and had a significant antitumor effect.

Objective To investigate the diagnosis,treatment and prognosis of isolated epidural hematoma (EDH) in children.Methods Clinical data of 83 children,45 males and 38 females (at age of 0.7-14 years),with EDH treated between January 2012 and October 2014 were retrospectively reviewed.Slipping and falling were the most commonly causes of injury.While motor-vehicle accident was noted with increased age.In 16 patients lesion of hematoma was frontal,in 28 patients temporal,in 13 patients parietal,in 9 patients occipital and in 17 patients posterior fossa.Results Of the 28 patients treated surgically,the outcome evaluated using the Glasgow Outcome Score (GOS) was good in 22 patients,mild disability in 2,moderate and severe disability in 1 and death in 3 (one died of venous sinus rupture and one large area infarction).Of the 55 patients treated conservatively,the outcome was good in 52 patients,mild disability in 1,and death in 2 (both presented with bilateral cerebral hernia on admission and one of them was unable to breath spontaneously).Conclusions Overall prognosis of the patients with isolated EDH in children is excellent.Complications of venous sinus rupture,foramen magnum hernia and large area cerebral infarction may contribute to poor prognosis.

Objective To investigate the diagnostic significance of synchronous observation on bone marrow biopsy,bone marrow smear and peripheral blood smear in stubborn hematonosis.Methods Thirty-seven patients with difficult and complicated hematologic diseases were detected by peripheral blood smear,bone marrow smear and bone marrow biopsy.Those slides and smears were observed.Results The 37 cases with blood disease was diagnosed of aplastic anemia in 3 cases,5 cases of myelodysplastic syndrome,6 cases of primary myelofibrosis,13 cases of secondary myelofibrosis,7 cases of lymphoma with bone marrow infiltration and 3 cases of metastatic carcinoma of bone marrow.In the diagnosis of aplastic anemia,myelodysplastic syndrome,primary and secondary myelofibrosis,lymphoma and metastatic carcinoma of bone marrow diagnosis,bone marrow biopsy is superior to smear.Conclusion Bone marrow biopsy,bone marrow smear and peripheral blood smear synchronous observation can effectively increase the diagnostic rate,decrease misdiagnosis rate,and correctly identify the stage of the disease.

Objective To evaluate the effect of surgical trauma on the cognitive function and expression of hepcidin and ferroportin 1 (FP1) in hippocampus in aged rats.Methods One hundred male Sprague-Dawley rats,aged 18 months,weighing 400-500 g,were randomly divided into 2 groups with 50 rats in each group:control group (group C) and surgical trauma group (group ST).The rats were anesthetized with chloral hydrate,but underwent no operation in group C.The rats Were anesthetized with chloral hydrate and underwent 30 min of modified exploratory laparotomy in group ST.Ten rats were chosen from each group at 24 h after operation and the cognitive function was assessed using Morris water-maze test for 6 consecutive days.Ten rats were sacrificed on 1st,3rd,5th and 7th days after beginning of Morris water-maze test and brains were removed for determination of hepcidin and FP1 expression in hippocampus by PCR and Western blot.Results Compared with group C,the escape latency was significantly prolonged,the time of staying at the original platform quadrant and frequency of crossing the original platform were decreased on 3rd,4th and 5th days after beginning of Morris water-maze test,and the expression of hepcidin was up-regulated and the expression of FP1 was down-regulated at each time point in group ST (P ＜ 0.05).Conclusion Surgical trauma can decrease the cognitive function in aged rats and the mechanism may be related to up-regulation of hepcidin expression and down-regulation of FP1 expression in hippocampus.

Cephalosporins are widely used antibiotics owing to their broad activity spectra and low toxicity. Many of these medically important compounds are made chemically from 7-aminodeacetoxycephalosporanic acid. At present, this intermediate is made by synthetic ring-expansion of the inexpensive penicillin G to form G-7-ADCA, followed by enzymatic removal of the side chain to obtain 7-ADCA. The chemical synthetic process is expensive, complicated and environmentally unfriendly. Environmentally compatible enzymatic process is favorable compared with chemical synthesis. In our previous research, metabolic engineered Escherichia coli strain (H7/PG15) was constructed and used as whole-cell biocatalyst for the production of G-7-ADC with penicillin G as substrate. The whole-cell biocatalysis was studied by single factor experiment, including the composition of substrates and the conversion conditions (OD600, pH, concentration of penicillin G, MOPS, glucose, time and FeSO4). After optimization, 15 mmol/L of G-7-ADCA was obtained. The process is convenient, efficient and economic. This work would facilitate the industrial manufacturing and further product research.
Anti-Bacterial Agents ; biosynthesis ; Biocatalysis ; Cephalosporins ; biosynthesis ; Escherichia coli ; metabolism ; Metabolic Engineering

Abstract: Objective　To investigate the diagnostic value of joint detection of Mycobacterium tuberculosis rifampicin resistance gene (Xpert MTB/RIF), Mycobacterium tuberculosis ribonucleic acid (TB-RNA) and Mycobacterium tuberculosis deoxyribonucleic acid (TB-DNA) in bronchoalveolar lavage fluid for smear-negative pulmonary tuberculosis. Methods A total of 806 patients with suspected smear-negative pulmonary tuberculosis admitted to our hospital from May 2020 to July 2022 were selected, 506 patients diagnosed as bacterial negative pulmonary tuberculosis by clinical, X-ray and sputum samples were classified as bacterial negative pulmonary tuberculosis group, and the other 300 patients with non-tuberculous pulmonary disease were classified as non-tuberculous pulmonary disease group. XpertMTB/RIF, TB-RNA and TB-DNA in bronchoalveolar lavage fluid of all patients were detected. With clinical, X-ray and sputum specimen examination of mycobacterium tuberculosis as the gold standard, the diagnostic efficacy of alveolar lavage solution Xpert MTB/RIF, TB-RNA and TB-DNA alone and in combination was analyzed. Results The positive detection rates of Xpert MTB/RIF, TB-RNA and TB-DNA in bronchoalveolar lavage fluid of the smear-negative pulmonary tuberculosis group and the non-tuberculosis pulmonary disease group were 69.96% (354/506) and 2.67% (8/300), 61.46% (311/506) and 5.00% (15/300), and 63.64% (322/506) and 8.00% (24/300), respectively. The rates in the smear-negative pulmonary tuberculosis group were higher than those in the non-tuberculosis lung disease group, and the differences were statistically significant (χ2=342.005, 246.930, 235.687, P<0.01). Compared with the gold standard, the sensitivity, specificity, accuracy, positive predictive value and negative predictive value of Xpert MTB/RIF in the diagnosis of smear-negative pulmonary tuberculosis were 69.96%, 97.33%, 80.15%, 97.79% and 65.77%, respectively; those values of TB-RNA were 61.46%, 95.00%, 73.95%, 95.40% and 59.38%, respectively; those values of TB-DNA were 63.64%, 92.00%, 74.19%, 93.06% and 60.00%, respectively; those values of combined diagnosis with Xpert MTB/RIF, TB-RNA and TB-DNA were 61.26%, 100.00%, 75.68%, 100.00% and 60.48%, respectively; the specificity and positive predictive value of combined detection were higher than those of single detection (P<0.05). Conclusions The joint detection of Xpert MTB/RIF, TB-RNA and TB-DNA in bronchoalveolar lavage fluid can improve the diagnostic efficacy of smear-negative pulmonary tuberculosis and is worthy of clinical promotion and application.