Objective To express the molecular chaperone Acr2 protein of Mycobacterium tuberculosis(Mtb)in E.coli and analyze the function. Methods The recombinant plasmid pET-28a-Acr2 was transformed into competent E. coli BL21(DE3),and induced by IPTG. The expressed His-Acr2 protein was purified by Ni-NTA chromatography and SuperdexTM200 10/300 GL gel filtration chromatography to obtain Acr2 protein. The Acr2 protein was refolded by spontaneous refolding and reassembly after thermal denaturation(100 ℃ for 15 min)and chemical denaturation(8 mol/L urea,37 ℃ for 4 h).The secondary structure of Acr2 protein before and after denaturation-renaturation was detected by circular dichroism spectroscopy and non-denaturing SDS-PAGE,and the molecular chaperone function of Acr2 protein in vitro was detected by substrate binding assay. Results The purified Acr2 protein had the relative molecular mass of about 232 000,the purity of over 90%,and the concentration of about 2 mg/mL,which recovered its natural secondary structure after denaturationrenaturation,and formed stable complexes with the denatured malate dehydrogenase(MDH)at 48 ℃. Conclusion The Acr2protein can restore its natural molecular conformation with molecular chaperone activity in vitro after denaturation-renaturation treatment,providing a new strategy for the preparation of Mtb protein antigen with natural activity.

Objective:To fabricate the composite scaffolds for bone regeneration with silk fibroin (SF), bacterial cellulose nanofibers (BCNR) and hydroxyapatite (HAp) and evaluate their osteogenic activity.Methods:HAp particles, BCNR and bone morphogenetic protein-2 (BMP2) were added into SF aqueous solution in turn, poured into molds of different sizes after being mixed evenly and processed at -25 ℃ for 24 hours to obtain frozen molds, and the composite scaffolds were frozen-dried by freezing-drying machine. The composite scaffolds with different mass ratios of SF and BCNR were divided into groups A (2∶1), B (4∶1) and C (6∶1), and the inactive composite scaffolds without BMP2 fell into group D. The surface morphology and pore structure of the scaffolds were detected by scanning electron microscopy. The porosity of the scaffolds was measured by mercury intrusion porosimeter. The stress-strain curve was obtained by using the universal material testing machine to compress the scaffolds, with which their compressive strength and Young′s modulus were analyzed. Immortalized mouse embryonic fibroblasts (iMEF) were inoculated on the composite scaffolds of group A, B, C and D. At 4 and 8 days after cell inoculation, the proportion of alive and dead cells in each group was detected by cell survival/death staining; the cell counting kit-8 (CCK-8) was used to detect cell proliferation activity in each group; the positive staining cells were detected in each group by alkaline phosphatase (ALP) staining; the ALP activity was observed in each group with ALP activity detection. A total of 15 female SD rats were selected to establish osteogenesis models with ectopic muscle bag. The composite scaffolds implanted with different SF/BCNR mass ratios and the inactive composite scaffolds without BMP2 fell into group A′ (2∶1), B′ (4∶1), C′ (6∶1) and D′ respectively, and a sham operation group was set at the same time, with 3 rats in each groups. In the sham operation group, the muscle bag and skin were sutured without scaffold implantation after the incision of skin, the blunt separation of the quadriceps muscle, and the formation of muscle bag in the muscle. In the other four groups, the corresponding scaffolds were implanted in the muscle bag and the muscle bag and skin were sutured. X-ray examination was performed at 2 and 4 weeks after operation to observe the osteogenesis in each group. At 4 weeks after operation, the implanted scaffolds and tissue complexes were collected by pathological tissue sectioning, HE staining and Masson staining, and for observing the osteogenesis by in each group. Immunohistochemical staining was also performed on the tissue sections to observe the expression of osteogenic markers type I collagen (COL1) and osteopontin (OPN) in each group.Results:Scanning electron microscopy showed that the lamellar and micropore structures of group B were more regular and uniform than those of groups A and C. The porosity rate analysis showed that the porosity rates of groups B and C were (89.752±1.866)% and (84.257±1.013)% respectively, higher than that of group A [(81.171±1.268)%] ( P<0.05 or 0.01), with the porosity rate of group C lower than that of group B ( P<0.01). The mechanical property test showed that the compressive strengths of groups B and C were (0.373±0.009)MPa and (0.403±0.017)MPa respectively, higher than that of group A [(0.044±0.003)MPa] ( P<0.01), and the Young′s moduli of groups B and C were (7.413±0.094)MPa and (9.515±0.615)MPa respectively, higher than that of group A [(1.881±0.036)MPa] ( P<0.01), with the compressive strength and Young′s modulus of group C higher than those of group B ( P<0.05 or 0.01). The cell survival/death staining showed that the number of dead cells of group B was significantly smaller than that of groups A, C and D at 4 days after cell inoculation, and that group B had the most living cells and the fewest dead cells at 8 days after cell inoculation. The results of CCK-8 experiment showed that at 4 days after cell inoculation, the cell proliferation activity of groups A and B was 0.474±0.009 and 0.545±0.018 respectively, higher than 0.394±0.016 of group D ( P<0.01); the cell proliferation activity of group C was 0.419±0.005, with no significant difference from that of group D ( P>0.05), while the cell proliferation activity of groups A and C were both lower than that of group B ( P<0.01). At 8 days after cell inoculation, the cell proliferation activity of group B was 1.290±0.021, higher than 1.047±0.011 of group D ( P<0.01); the cell proliferation activity of group C was 0.794±0.032, lower than that of group D ( P<0.01); the cell proliferation activity of group A was 1.086±0.020, with no significant difference from that of group D ( P>0.05); the cell proliferation activity of groups A and C was lower than that of group B ( P<0.01). At 4 and 8 days after cell inoculation, ALP staining showed that more positive cells were found in groups A, B and C when compared with group D, and more positive cells were found in group B than in groups A and C. At 4 days after cell inoculation, the ALP activity detection showed that the ALP activity of groups A, B and C was 1.399±0.071, 1.934±0.011 and 1.565±0.034 respectively, higher than 0.082±0.003 of group D ( P<0.01), while the ALP activity of groups A and C was lower than that of group B ( P<0.01). At 8 days after cell inoculation, the cell activity of groups A, B and C was 2.602±0.055, 3.216±0.092 and 2.145±0.170 respectively, higher than 0.101±0.001 of group D ( P<0.01), while the ALP activity of groups A and C was lower than that of group B ( P<0.01). X-ray examination results showed that at 2 weeks after operation, no obvious osteogenesis was observed in the sham operation group, group D′, A′ and C′, while it was observed in group B′. At 4 weeks after operation, obvious osteogenesis was observed in group A′, B′ and C′, with significantly more osteogenesis in group B′ than in the other two groups, while there was no obvious osteogenesis in the sham operation group and group D′. At 4 weeks after operation, the HE staining and Masson staining showed that a large number of uniformly distributed new bone tissue was formed in group B′, while only a small amount of new bone tissue was found locally in groups A′ and C′, and only part of new tissue was found to grow in group D′ with no obvious new bone tissue observed. The maturity of new bone tissue formed in group B′ was higher than that in group A′ and C′. Immunohistochemical staining showed more COL1 and OPN positive staining in group B′ when compared with groups A′ and C′. The expression intensity analysis of COL1 and OPN showed that in groups A′, B′ and C′, the expression intensity of COL1 was 2.822±0.384, 22.810±2.435 and 12.480±0.912 respectively and the expression intensity of OPN was 1.545±0.081, 5.374±0.121 and 2.246±0.116 respectively, with higher expression intensity of COL1 and OPN in groups B′ and C′ than that in group A′ ( P<0.01) and lower expression intensity of COL1 and OPN in group C′ than that in B′ group ( P<0.01). Conclusions:The composite scaffold for bone regeneration is successfully fabricated with SF, BCNR and HAp. The composite scaffold with a mass ratio of SF to BCNR of 4∶1 has uniform pore structure, high porosity, good mechanical properties and biocompatibility, excellent pro-osteogenic properties in vitro, as well as excellent osteo-inductivity and osteo-conductivity.

The standardization of classification methods of Traditional Chinese Medicine(TCM) ancient books can provide a clear and reliable reference for all kinds of TCM ancient books collection units, which can also promote the sharing and utilization of TCM ancient books. We studied and investigated the classification methods of TCM ancient books in past dynasties. The standard on classification of TCM ancient books was formulated by compared with the classification table of Zhongguo Zhongyi Guji Zongmu, and referred to the classification table of Zhonghua Guji Zongmu. This standard specified three-level categories and classification principles of TCM ancient books, and mainly composed of basic categories, three-level category table, classification principles and examples, and instructions for use.

Objective:To explore the epidemiological characteristics of geriatric hip fractures in Beijing so as to provide evidence for effective prevention and control measures.Methods:This multicenter study was conducted in 3 urban (Beijing Jishuitan Hospital, Beijing Hospital and Beijing Anzhen Hospital) and 3 suburban hospitals (Beijing Shunyi District Hospital, Beijing Changping District Hospital and Beijing Liangxiang Hospital) in Beijing from November 2018 to November 2019. Eligible patients were those aged ≥ 65 years with hip fracture confirmed by X-ray and being admitted to hospital within 21 days of injury. To explore the epidemiological characteristics of geriatric hip fractures in Beijing, such data were collected as patients' age, gender, comorbidities, as well as type, site, time and cause of the fracture.Results:① A total of 2,071 patients were included in this suevey. They were 653 males and 1,418 females (M∶F=1∶2.17). Their age ranged from 65 to 102 years (average, 79.8 years). The patients aged from 75 to 84 years were the most common, accounting for 44.81% (928/2,071). ② Femoral neck fractures accounted for 43.41% (899/2,071), and intertrochanteric fractures accounted for 56.59% (1,172/2,071). The age of the patients with femoral neck fracture was (78.6±7.7) years, which was significantly younger than that of those with intertrochanteric fracture [(80.7±7.4) years] ( P<0.05). ③ 94.69% of the hip fractures (1,961/2,071) were caused by falling, and 71.27% fractures (1,476/2,071) happened at home. ④ Approximately 83.00% of the patients (1,719/2,071) had one or more comorbid conditions. Hypertension was the most prevalent disease (57.89%, 1,199/2,071), followed by diabetes (27.09%, 561/2,071), and coronary atherosclerotic heart disease (22.02%, 456/2,071). Conclusions:In Beijing, significantly more geriatric females sustain a hip fracture than males, and the proportion of those aged from 75 to 84 year is the largest. The proportion of intertrochanteric fractures increases with age. Falls are the leading cause for geriatric hip fractures. Most of the patients have one or more chronic comorbid conditions. Corresponding prevention and intervention measures should be formulated according to the distribution characteristics of elderly hip fractures in Beijing.

Objective To analyze the antibiotic resistance of Branhemella catarrhalis strains isolated from sputum specimens of patients with lower respiratory tract infections from Linyi, Shandong Province, and to explore the relationship between bro genotypes of the strains and their resistance to antibiotic agents.Methods Sputum specimens were colleted from the patients with lower respiratory tract infections in Linyi People ’ s Hospital from the January 2010 to December 2014.The specimens were inoculated into 4 different disks for bacterial isolation and cultivation.β-lactamase detection and drug sensitivity tests were performed, and PCR coupled with restriction endonuclease analysis was employed for bro genotyping.χ2 test was used to compare drug resistance of strains with different bro genotypes.Results A total of 497 Branhemella catarrhalis strains were isolated in five years, among which 221 strains were isolated in winter.All strains were sensitive to ertapenem and chloramphenicol, and the resistance rates to amoxicillin/clavulanate and cefaclor were low (≤2.8%).The strains were highly resistant to compound sulfamethoxazole, erythromycin and ampicillin (47.6%-89.8%), and there was a trend of increasing resistance rates with the year, but no statistically significant difference was observed ( P >0.05 ) .β-lactamases was positive in 412 strains (82.9%), and all of these strains were positive for bro gene, and the resistances to erythromycin, compound sulfamethoxazole, levofloxacin and ampicillin were higher in bro positive strains than those in bro negative strains (χ2 =12.16, 16.18, 8.41 and 200.00,P<0.05).Among bro positive strains, 391 (94.9%) were of genotype bro-1, 21 (5.1%) were of genotype bro-2, and their resistance to antibiotic agents was not of statistical difference ( P >0.05 ).Conclusions Most of Branhemella catarrhalis clinical isolates are β-lactamase producing strains, and bro-1 is the most common genotype.Strains are highly sensitive to carbapenems, cephalosporins andβ-Lactamaseinhibitors, which can be recommended for the treatment of Branhemella catarrhalis-related respiratory tract infections.

OBJECTIVETo investigate the effects of Naokang Erhao decoction on the cognitive ability and the expression of Caspase-3, Bax and Bcl-2 proteins in hippocampus of type 2 diabetic rats.

METHODThe diabetes mellitus (DM) rat model was produced by intraperitoneal injection of streptozotocin and fed with high fat and sucrose diet. The Naokang Erhao-treated rats were intragastrically given different doses of Naokang Erhao, whereas the control and DM model groups were given double distilled water for 4 consecutive weeks. Learning and memory abilities of rats were tested with the Morris water maze. The expression of Caspase-3, Bax and Bcl-2 proteins in hippocampal CA1 region was measured by immunohistochemistry.

RESULTBoth escape latency and swimming distance of type 2 DM rats were significantly prolonged in comparison of those in normal control (P < 0.01), and swimming time in the platform of previous quadrant was significantly shorter in model group (P < 0.01). Furthermore, the expression of Bcl-2 protein was decreased, while Caspase-3 and Bax in the hippocampus were increased compared with the control group (P < 0.01). Four weeks of treatment with Naokang Erhao decoction remarkably improved the learning and memory abilities of DM rats, increased the expression of Bcl-2 and decreased the expression of Caspase-3 and Bax in hippocampal CA1 region of model rats (P < 0.01 or P < 0.05).

CONCLUSIONNaokang Erhao decoction may inhibit apoptosis by increasing the expression of Bcl-2 and reducing the expression of Caspase-3 and Bax in the hippocampus, and this may be one of the mechanisms by which Naokang Erhao decoction improves cognitive ability in DM rats.

Acetophenones ; pharmacology ; Animals ; Apoptosis ; drug effects ; Benzoates ; pharmacology ; Bridged-Ring Compounds ; pharmacology ; CA1 Region, Hippocampal ; drug effects ; metabolism ; Caspase 3 ; drug effects ; metabolism ; Cognition Disorders ; drug therapy ; metabolism ; Diabetic Neuropathies ; drug therapy ; metabolism ; Diterpenes, Abietane ; pharmacology ; Drug Combinations ; Drugs, Chinese Herbal ; pharmacology ; Glucosides ; pharmacology ; Male ; Maze Learning ; drug effects ; Proto-Oncogene Proteins c-bcl-2 ; drug effects ; metabolism ; Pyrazines ; pharmacology ; Rats ; bcl-2-Associated X Protein ; drug effects ; metabolism

Objective To study the level of serum ICAM-1, VCAM-1, E-selectin, P-selectin, IL-6 and CRP in different Chinese medicine syndrome patients with carotid artery atherosclerosis. Methods Blood sample were collected from 176 patients with carotid artery atherosclerosis. The concentration of ICAM-1, VCAM-1, E-selectin, P-selectin, IL-6 and CRP were tested by means of ELISA. The subjects were divided into 3 groups according to the Chinese medicine syndrome, and the concentrations of inflammatory factors were compared among the 3 groups. Results Compared with control group, the level of inflammatory factors in the patients with carotid atherosclerosis was higher, and the levels of ICAM-1 and VCAM-1 in phlegm group were higher than blood stasis group and marrow deficiency group (P

Objective To analyze the internal relationship between phlegm stasis syndrome (痰浊内阻证) and blood stasis syndrome (血瘀证) in cases with carotid atherosclerosis. Methods The clinical data of 136 patients with atherosclerosis accompanied by phlegm stasis and blood stasis syndromes were collected from the investigation table of the 4 diagnostic methods in traditional Chinese medicine(TCM); the principal components of the clinical data were statistically analyzed, and the correlative relationships of these principal components were studied. Results There were two principal components of phlegm stasis syndrome, which were tz1 and tz2. And tz1 cumulated 42.645% to phlegm stasis syndrome, which could be considered as phlegm stasis in limbs and tunnel; tz2 cumulated 24.898% to phlegm stasis syndrome, which could be considered as phlegm stasis in the body that might impair the 7 orifices (such as eyes, ears, nose and mouth). And they could also be considered as special markers of phlegm stasis syndrome of carotid atherosclerosis. There were three principal components of blood stasis syndrome, which were xy1, xy2 and xy3. And xy1 cumulated 37.197% to blood stasis syndrome, which could be considered as blood stasis in the artery and tunnel; xy2 cumulated 21.627% to blood stasis syndrome, which could be considered as the blood outside the vessels located in the body and could obstruct tunnel and could not nourish muscle and skin; then, xy3 cumulated 13.685% to blood stasis syndrome, which could be considered as the blood stayed in the brain and could not nourish brain and might reflect as a special marker of blood stasis syndrome of carotid atherosclerosis. There was significant positive correlative relationship between phlegm stasis syndrome principal component 1 and blood stasis syndrome principal component either 1 or 2 (P

OBJECTIVE To investigate the distribution and resistance of Pseudomonas aeruginosa isolated during Jan-Dec 2006.METHODS The clinically isolated P.aeruginosa strains were collected cultured and identified by paper diffusing method or trace dilution method(MIC),the results were evaluated according to the relevant documents of NCCLS of USA.RESULTS The in vitro susceptibility test of 244 P.aeruginosa isolates to 16 kinds of antibacterials indicated the resistance rate to SMZ was the highest(98.8%);then to minocycline,tetracycline and ticarcillin/clavulanicacid(70.1%,58.6% and 54.5% respectively).CONCLUSIONS To strengthen the continuous survezillance of drug resistance of P.aeruginosa,to sum up the resistance rules of main pathogens of departments in hospital and to reduce of production of resistant bacterica have the important significance.

Objective: To analyses the traditional Chinese medicine syndromes of chronic fatigue syndrome(CFS),by the way of variable cluster analysis.Methods: We researched 237 CFS patients,and recorded their symptoms,tongues and pulses.Then we used cluster analysis way to analyses these patients' clinical data.Results: These patients could be divided into 4 types: marrow deficiency syndrome,yin fluid deficiency syndrome,yang deficiency of spleen and kidney syndrome and overabundant liver-fire syndrome.The variable proportion is 61.68%.Conclusion: CFS is mainly produced by deficiency and/or excessive in traditional Chinese medicine pathology mechanism.And,the way of variable cluster analysis could help CFS patients' TCM syndrome differentiation.