Objective To summarize the clinical presentations, the findings of lab tests and procedures and the genetic investigation of collagen type Ⅵ related myopathy, and to help clinicians recognize and diagnose this rare disease.Methods Seven familiar or spontaneous collagen type Ⅵ related myopathy patients diagnosed by gene detection were analyzed.We emphasized on the features of clinical manifestations, serum creatine kinase level, electromyography, lower-limb muscle MRI, muscle biopsy and correlation between genotype and pZenotype.Results Among 7 patients, 3 were caused by COL6A1 mutation, 1 was caused by COL6A2 mutation and 3 were caused by COL6A3 mutation.Two patients were familiar wZile 5 were spontaneous.HigZligZted clinical presentations were proximal weakness in lower limbs and joint contrature.Serum creatine kinase level was sligZtly elevated.ElectromyograpZy sZowed sligZt myogenic damage.Muscle MRI of tZigZ sZowed distinct pattern of muscle involvement.Muscle patZology revealed dystropZic myogenic cZanges with proliferation of connective tissue between muscle fibers.Conclusions Neurologists should recognize the features of collagen type Ⅵ related myopathy, such as progressive weakness, early-onset joint contraetures, slightly elevated serum creatine kinase and selective muscle involvement on leg MRI scan, and then perform next-generation sequencing based genetic test on suspected patients.This approach would improve the diagnostic rate of the disease.

To improve 3-ketosteroid-△1-dehydrogenase(KSDH) activity and the transformation level for androst-4-ene-3,17-dione, 3-ketosteroid-△1-dehydrogenase gene(ksdD) from Arthrobacter simplex was cloned into plasmid pWB980 and expressed in B. subtilis WB600 under the control of promoter P43. The molecular weight of expressed enzyme was about 55kDa by SDS-PAGE analysis. The activitities assayed by spectrophotometrical method of intracellular and extracellular soluble enzyme were 110?0.5mU and 15?0.6mU per milligram of protein respectively. The transformation rate of androst-4-ene-3,17-dione by the B. subtilis recombinant cells was 45.3%. Compared with Arthrobacter simplex, the enzyme activity of KSDH expressed in B. subtilis was improved about 30 fold, and the transformation level of androst-4-ene-3,17-dione by the B. subtilis recombinant cells was improved about 10 fold. The recombinant B. subtilis cells used in biotransformation of steroids provided a new way for steroid medicines production.

Objective To investigate the imaging findings and key diagnostic points of Brunner’s gland adenoma on CT.Methods The CT imaging findings and pathological features of 9 cases of Brunner’s gland adenoma confirmed by pathology were retrospec-tively analyzed,including the lesions number,site,size,shape,margin,density and the enhancement pattern.Results Of the 9 ca-ses,1 case located in the antrum and 8 cases in the duodenaum [6 cases in the duodenal bulb (75%)and 2 in the papillary (25%)]. Of the 6 cases of duodenal lesions,3 were found at the anterior wall and 3 at the posterior wall.Except 1 case which complicated with enteritis and had an obscure margin,the other 8 cases were clear margined ,and were round or nodular in shape.The maximum size of tumors ranged 1 5-68 mm in diameters (mean 35.0 mm ± 1 6.2 mm).The density of tumors was homogeneous on CT scan without necrosis or hemorrhage.In the arterial phase after administration of contrast agent,the lesions were similar to the adjacent intestinal wall enhancement,and mucosal annular enhancement (halo sign)showed in 6 cases,and the dot-shape non-enhancement area within the lesion (black star sign)showed in 5 cases,and the thickening or tortuous enhanced blood vessel showed in 6 cases.In the venous phase,9 cases were progressive enhancement,and the “black star sign”or “the black line sign”showed more clearly in 5 cases.In the pathology,the lesions were polypoid-like,solid or cystic.Under the microscope,the hyperplasic Brunner’s glands were covered with normal duodenal mucosa and separated by bundles of smooth muscle cells with dilated duct,cyst,and adipose cells,1 case with atypical hyperplasia of the glandular epithelium and 1 case with ectopic pancreas.Conclusion There are some spe-cific CT imaging features in Brunner’s gland adenoma,which is of important clinical value in accurate preoperative diagnostic.

Objective To establish an animal model of severe blast lung injury in baby rabbits,and to provide a way to study the char-acteristic and treatment of blast lung injury in minors.Methods Randomly selected sixteen 4-weeks old New Zealand white rabbits,and the blast lung injuries were made by BST-Ⅰ biological shock tube with different drive pressure (4.0 MPa and 4.5 MPa)respectively.Then compared the injury severity of the 4.0 Mpa group and the 4.5 MPa group.Selected forty-eight 4-weeks old New Zealand white rabbits and di-vided them into the control group (8 rabbits)and the blast lung injury group (40 rabbits)Rabbits in the blast lung injury group were injured with 4.5 MPa drive pressure.Observed the vital signs,physiological index,gross anatomy of the lung,pathology,and pulmonary water content at the time of injury immediately (0 hour),2 hours,4 hours,6 hours,12 hours,24 hours,48 hours and 72 hours after the injury.Results Rabbits inthe 4.0 Mpa group and the 4.5 MPa group were all alive.The overpressure of blast wave of the 4.0Mpa group was (328.16 ± 4.78)kPa,rate of severe pulmonary defense was 12.5%,and the AIS score was (3.38 ±0.52)points.In the 4.5 MPa group,the overpressure of blast wave was (395.04 ±11.74)kPa,rate of severe pulmonary defense was 87.5%,and the AIS score was (4.13 ±0.64) points.Rabbits in the control group and the blast lung injury group were all alive.The spirits of rabbits were drooping immediately after inju-ry,and it last about 0.5 hour.Then the breathing and heart rate was accelerated,pulmonary water content was increased significantly,and there were extensive hemorrhage and edema in the lung.Most of the rabbits suffered severe lung injury,and the AIS score was (3.98 ±0.55) points.Lung tissue rupture,hemorrhage,edema,and inflammatory cells infiltration were the main pathological manifestations under light microscopy. Conclusion The model of severe blast lung injury in baby rabbits could be established with BST-Ⅰbiological shock tube and drive pressure of 4.5 MPa.It is relatively simple,easily controllable and highly repeatable,which can be used as a feasible model for the study of blast lung injury.

Background Researches documented that retinal nerve fiber layer thickness (RNFLT) in unaffected carriers of Leber hereditary optic neuropathy (LHON) becomes thickened in different quadrants to different degrees.But the change of their macular thickness is still unclear.Objective This study was to clarify RNFLT and macular thickness by optical coherence tomography (OCT) in unaffected female carriers of LHON families.Methods Five female LHON patients (5 eyes) from 5 LHON families,eighteen unaffected female carriers (18eyes) from 18 LHON families and twenty-five age-matched healthy female controls (25 eyes) were included in this study.The patients and genetic carriers were diagnosed in PLA General Hospital from 2011 September to 2012 October.Regular ocular examination were performed followed by OCT measurement of retinas.The Optic Disc Cube 200×200 and Macular Cube 200×200 protocols were used during the OCT measurement.Average (360°) RNFLT,RNFLT at four quadrantic sections,cube average macular thickness and macular thickness of nine Early Treatment Diabetic Retinopathy Study (ETDRS) sub-areas were compared among the LHON genetic carriers,LHON patients and normal controls.Results Compared to the normal control group,significant reduced values were seen in temporal,superior,nasal and inferior side of sub-area macular thickness in the LHON female carriers (P=0.022,0.046,0.024,0.008).In addition,but no significant differences were found in cube average thickness,central subarea macular thickness,temporal,superior,nasal and inferior side of lateral sub-area macular thickness,average RNFLT,and temporal,superior,nasal and inferior quadrant RNFLT between the LHON female carriers and normal controls (P=0.102,0.051,0.238,0.663,0.1 10,0.104,0.419,0.371,0.158,0.063,0.563).Compared to the unaffected female carrier group,female patients showed significant reductions in cube average macular thickness,temporal,superior,nasal and inferior side of sub-area macular thickness,temporal,superior,nasal and inferior side of lateral sub-area mac ular thickness,average R NFLT and temporal,superior,and inferior quadrant RNFLT (P =0.000,0.000,0.000,0.007,0.002,0.002,0.000,0.000,0.040,0.000,0.016,0.000,0.000) except for the central subarea macular thickness and nasal quadrant RNFLT (P=0.388,0.580).Conclusions Unaffected LHON female carriers show a normal peripapillary RNFLT,but the macular thickness at medial sub-area is thinner.This first report offers an information of macular structure change in unaffected LHON female carriers,which suggest that macular damage appears prior to RNFLT change.

Objective:To investigate the expression of circhipk3 in microglial cells in heat-induced neurological injury, and to preliminary analyze the effect of circhipk3 on microglial polarization in heat-induced neurological injury.Methods:Mice were randomly (random number) divided into a control group and a heat radiation disease 0.8 h group (HS 0.8), a heat radiation disease 8h group (HS 8), and a heat radiation disease 24 h group (HS 24). By establishing a mouse model of heat shock (HS), heat-damaged brain tissue was obtained, microglia were isolated and RNA was extracted. Quantitative PCR method was used to detect M1 and M2 marker molecules in microglia, and to evaluate the polarization direction and type of microglia. The expression level of circhipk3 was detected in microglial cells in heat-induced neurological injury, and the effect of circhipk3 on microglial polarization was further elucidated by intervening the expression of circhipk3 in microglial cells.Results:The expression of CD45 and CD11-b in the HS 8 group was significantly higher than that in the control group [(4.41±0.18) vs. (1±0.15), P=0.000], [(3.47±0.19) vs (1±0.15), P=0.000] , and the CD45 and CD11-b of the HS 24 group was significantly lower than that of the HS 8 group [(1.34±0.15) vs. (4.41±0.18), P=0.000], [(1.38±0.21) vs. (3.47±0.19), P= 0.001]. At the same time, the expression of CD206, FIZZ and Arg1 in the HS 8 group started to increase compared with the control group [(1.59±0.16) vs. (1±0.12), P=0.014], [(1.62±0.15) vs. (1±0.15), P=0.002 ], [(2.23±0.28) vs. (1±0.19), P=0.004], and CD206, FIZZ, and Arg1 in the HS 24 group were significantly higher than those in the control group [(2.67±0.20) vs. (1±0.12), P=0.002], [(2.19±0.15) vs. (1±0.15), P=0.000], [(3.04±0.18) vs. (1±0.19), P=0.001]; circhipk3 mimicis significantly increased the expression of Arg1 [(7.26± 0.06) vs. (3.86±0.06), P=0.000]; at the same time, circhipk3 inhibitor promoted the expression of CD45 and HO-1 [(2.96±0.03) vs. (1.63±0.09), P=0.000], [(2.52±0.10) vs. ( 1.30±0.02), P=0.000]. Conclusions:Microglial cells are predominantly M1-type in early neurological injury of heat radiation disease. HO-1 may be one of the microglial M1-type markers. The high expression of circhipk3 in microglial cells mainly promotes its transformation to M2 type.

At present, there is no cure for acquired immune deficiency syndrome (AIDS) due to HIV-1 latent reservoirs. Therefore, it urgently requires novel HIV-1 latency-regulating agents with high potency, low toxicity and favorable drug-like properties to achieve a functional cure for AIDS. Herein, we reviewed the advances in HIV-1 latency-regulating agents since 2019, including the drug discovery strategies, bioactivities, and mechanisms of these compounds. It is of great guiding significance in the development of latency-regulating agents with clinical value.

AIMTo establish a method for determinate of the inhibitory activity of angiotensin-converting enzyme inhibitor captopril by high performance capillary electrophoresis.

METHODSThe characteristic absorptive wavelength of hippuric acid determined by ultraviolet spectrophotometer is 228 nm. The method employed a melted capillary column, 50 mmol.L-1 phosphoric acid (pH 8.3) buffer solution, inject pressure 4.8 kPa, inject time 3 s, separation voltage 20 kV and detection wavelength 228 nm.

RESULTSThe reactant and resultant was separated completed within 7 min. IC50 of captopril was 0.019 mumol.L-1. Captopril is a competitive inhibitor, which was proved by enzyme reaction dynamics.

CONCLUSIONThe method was shown to be accurate, simple and rapid and can be used for determination of the inhibitory activity of captopril.

Angiotensin-Converting Enzyme Inhibitors ; pharmacology ; Captopril ; pharmacology ; Electrophoresis, Capillary ; methods ; Hippurates ; analysis ; Peptidyl-Dipeptidase A ; metabolism

To improve 3-ketosteroid-△1-dehydrogenase (KSDH) activity and the transformation level for androst-4-ene-3,17-dione,3-ketosteroid-△1 -dehydrogenase gene (ksdD) from Arthrobacter simplex was cloned into plasmid pWB980 and expressed in B. subtilis WB600 under the control of promoter P43. The molecular weight of expressed enzyme was about 55kDa by SDS-PAGE analysis. The activitities assayed by spectrophotometrical method of intracellular and extracellular soluble enzyme were 110 ± 0.5mU and 15 ± 0.6mU per milligram of protein respectively. The transformation rate of androst-4-ene-3,17-dione by the B. subtilis recombinant cells was 45.3%. Compared with Arthrobacter simplex, the enzyme activity of KSDH expressed in B. subtilis was improved about 30 fold, and the transformation level of androst-4-ene-3,17-dione by the B.subtilis recombinant cells was improved about 10 fold. The recombinant B. subtilis cells used in biotransformation of steroids provided a new way for steroid medicines production.

BACKGROUNDRepair of large bone defects remains a challenge for clinicians. The present study investigated the ability of mesenchymal stem cells (MSCs) and/or periosteum-loaded poly (lactic-co-glycolic acid) (PLGA) to promote new bone formation within rabbit ulnar segmental bone defects.

METHODSRabbit bone marrow-derived MSCs (passage 3) were seeded onto porous PLGA scaffolds. Forty segmental bone defects, each 15 mm in length, were created in the rabbit ulna, from which periosteum was obtained. Bone defects were treated with either PLGA alone (group A), PLGA + MSCs (group B), periosteum-wrapped PLGA (group C) or periosteum-wrapped PLGA/MSCs (group D). At 6 and 12 weeks post-surgery, samples were detected by gross observation, radiological examination (X-ray and micro-CT) and histological analyses.

RESULTSGroup D, comprising both periosteum and MSCs, showed better bone quality, higher X-ray scores and a greater amount of bone volume compared with the other three groups at each time point (P < 0.05). No significant differences in radiological scores and amount of bone volume were found between groups B and C (P > 0.05), both of which were significantly higher than group A (P < 0.05).

CONCLUSIONSImplanted MSCs combined with periosteum have a synergistic effect on segmental bone regeneration and that periosteum plays a critical role in the process. Fabrication of angiogenic and osteogenic cellular constructs or tissue-engineered periosteum will have broad applications in bone tissue engineering.

Animals ; Bone Regeneration ; physiology ; Cells, Cultured ; Lactic Acid ; chemistry ; Mesenchymal Stromal Cells ; cytology ; Periosteum ; cytology ; Polyglycolic Acid ; chemistry ; Rabbits ; Tissue Engineering ; methods ; Tissue Scaffolds ; chemistry