BACKGROUND:Tissue and cellimplantation entails high-quality seed cells. In order to satisfy this requirement, it is crucial to produce adequate wel-conditioned, high-purity and strong proliferation ability bone marrow-derived mesenchymal stem cells.
OBJECTIVE:To establish a simple, rapid and effective in vitro isolation and culture method of bone marrow-derived mesenchymal stem cells, and to define the biological features of bone marrow mesenchymal stem cells.
METHODS:Rat bone marrow mesenchymal stem cells were isolated from the bilateral tibial and femoral bones by the method of whole bone marrow, then purified and passaged by attachment method. The morphology and features of bone marrow mesenchymal stem cells were observed, the growth curve was drawn and the cellsurface antigen was detected by flow cytometry. The bone marrow mesenchymal stem cells were induced to differentiate along the osteogenic, chondrogenic and adipogenic lineages.
RESULTS AND CONCLUSION:Bone marrow mesenchymal stem cells isolated by the whole bone marrow adherence method grew vigorously and were highly purified. The cultured cells were spindle-shaped. The growth curve was S-shaped and the population doubling time was 29 hours. The cells stil maintained a strong proliferative capacity after they were passaged for 10 generations. The surface markers such as CD44, CD29, CD90 were positive, while CD45, CD34, CD11b were negative. At the third passage, bone marrow mesenchymal stem cells were induced to differentiate along the osteogenic, chondrogenic and adipogenic lineages, respectively. Fol owing induction, Alizarin red staining, alkaline phosphatase staining, von-kossa mineralized nodules staining, toluidine blue staining, and oil red O staining were al positive. This shows that the whole bone marrow adherence method is a simple and reliable method for the in vitro isolation, culture and proliferation of bone marrow mesenchymal stem cells. Moreover, they have multi-lineage differentiation capacity under different inducers. The third passage bone marrow mesenchymal stem cells have the highest biological activity and can act as the ideal seed cells for subsequent experiments.

Objective To explore the prevalence and influential factors of chronic respiratory system diseases among resi-dents over 1 5 years old in Hubei province and provide evidence for disease prevention.Methods During October to November in 2013,through stratified cluster sampling,we sampled 20 cities or counties.The survey included the the general condition of family, individual,chronic diseases.Results A total of 28 563 residents answered the questionnaire and 423 of them reported chronic re-spiratory system diseases by themselves.The prevalence rate was 14.8‰.These included 229 cases with COPD(54.1%),44 cases with asthma(10.4%),35 cases with chronic pharyngolaryngitis(8.3%)and 1 1 5 cases with other chronic respiratory system disea-ses(27.2%).In urban and rural area,the prevalence rate were 13.6‰ and 1 5.7‰ respectively.Multivariate logistic analysis showed that gender,age,economic status and medical insurance are influential factors of chronic respiratory system diseases.Conclusion Prevalence rate of chronic respiratory system diseases among residents over 1 5 years old in Hubei province was slightly increased and disease control measures should be brought out.

Objective : To investigate the willingness to receive HIV testing in primary health service institutions (PHSIs) among young students in Wuhan City, so as to provide the evidence for improving the detection of HIV testing among young students. Methods: Fifteen PHSIs were sampled using a stratified random sampling method in 14 districts of Wuhan City, and school students at ages of 15 to 24 years were sampled from each district using a convenience sampling method. Participants' demographics, awareness of AIDS-related knowledge, HIV testing and willingness to receive HIV testing were collected using questionnaires, and factors affecting the willingness to receive HIV testing in PHSIs were identified among school students using a multivariable logistic regression model. Results : A total of 301 questionnaires were allocated, and 299 valid questionnaires were recovered, with an effective recovery rate of 99.34%. The respondents included 143 men (47.83%) and 156 women (52.17%), and had a mean age of (19.36±2.40) years; there were 223 respondents with an educational level of diploma and above (74.58%). The awareness of AIDS-related knowledge was 71.57% among the respondents, and 144 respondents had received AIDS-related health education in PHSIs (48.16%). There were 34 respondents that had received HIV testing (11.37%) and 203 respondents that were willing to receive HIV testing in PHSIs (67.89%). The respondents that were unwilling to receive HIV testing in PHSIs were mainly attributed to considering to be unlikely to get HIV infections (82.29%). Multivariable logistic regression analysis showed that school students who knew AIDS-related knowledge (OR=2.797, 95%CI: 1.583-4.941), knew free HIV counseling and testing services in PHSIs (OR=2.070, 95%CI: 1.123-3.814), and had received AIDS-related health education in PHSIs (OR=2.814, 95%CI: 1.573-5.032) were more willing to receive HIV testing in PHSIs. Conclusions There were 67.89% of school students that were willing to receive HIV testing in PHSIs in Wuhan City, and the willingness to receive HIV testing was correlated with the awareness of risk of HIV infections, and awareness and experience of AIDS control services in PHSIs.

Objective To investigate the causes of peripheral cytopenia in patients with posthepatitic cirrhosis and portal hypertensive splenomegaly.Methods The clinical data of 183 patients with hepatitic cirrhosis and portal hypertensive splenomegaly complicated by peripheral cytopenia who were operated in our hospital in the past 17 years were retrospectively studied.Results All these patients underwent splenectomy.Before operation,all these patients had one or more types of peripheral cytopenia (cumulative cytopenia:390 patient-times).After splenectomy,blood counts in 79.2％ returned to normal;in 15.9％ increased but failed to reach normal levels;and in 4.9％ became lower than before operation.5 patients died soon after operation.Conclusion Hypersplenism is the main cause for the peripheral cytopenia most cirrhotic portal hypertension patients.Splenectormy is an effective method to treat hypersplenism.

Objective To investigate the difference in the microstructure of gray matter nucleus in different movement subtypes of Parkinson’s disease (PD) by diffusion kurtosis imaging ( DKI) technique, and to analyze the correlation with clinical manifestations. Methods Ninety-seven patients with PD and 83 healthy controls performed conventional MRI sequence and DKI sequence scan. The PD patients were classi-fied into gait disorder subtype (PIGD,n=57) and tremor dominant subtype (TD,n=40)subtypes according to motor symptoms. Fractional anisotropy (FA),mean diffusivity (MD),axial diffusivity (Da),radial diffu-sivity(Dr),mean kurtosis (MK),axial kurtosis (Ka) and radial kurtosis (Dr) maps and data were genera-ted by software after processing. DKI was performed for all subjects and data was collected from different brain regions in both hemispheres,including red nucleus(RN),substantia nigra pars reticulate( SNr),sub-stantia nigra pars compacta(SNc),putamen(PUT),globus pallidus(GP),head of caudate nucleus (CN)and thalamus(THA). Results TD showed a higher MMSE score(P=0. 019),but lower modified Hoehn-Yahr score than that in PIGD (P<0. 001),there was no significant difference of age of onset,sex,limbs of onset or disease duration between two PD subgroups. Compared with healthy controls, both TD and PIGD showed down-regulated MD,Da and Dr and up-regulated Ka values(P<0. 001); MK(0. 83±0. 26,0. 80±0. 18) was increased in SNr both in TD and PIGD,while SNc,PUT and GP (0. 84± 0. 20,0. 75± 0. 07,0. 81± 0. 14) were decreased only in TD (P=0. 017,P=0. 010,P=0. 020,P<0. 001,P=0. 002). The Kr values of PUT and CN(0. 71±0. 17,0. 72±0. 14) were reduced in PIGD,while CN(0. 70±0. 14) were reduced in TD re-spectively (P=0. 002,P=0. 031,P=0. 007). The MK was lower in TD than that in PIGD (t=-2. 214,P=0. 029),and no significant difference was found in other grey matter nuclei between TD and PIGD ( P>0. 05). Moreover,there was no significant correlation between DKI value and disease duration,MMSE score or Hoehn-Yahr scale (P>0. 05) in TD and PIGD. Conclusion There is heterogeneity of clinical symptoms between these two subgroups of PD. DKI can quantify the microstructural changes of grey matter nucleus in different type PD patient.

Objective:To investigate the effectiveness and advantage of visual laryngoscope in the treatment of patients with sudden cardiac arrest who need spontaneous respiration tracheal intubation.Methods:Totally 60 patients who suffered from cardiac arrest and needed spontaneous respiration tracheal intubation were enrolled from June 2020 to February 2021 in the Affilicated Hospital of Chifeng University. Theywere randomlydivided into two groups-traditional laryngoscope (TL) group and visual laryngoscope (VL) group, with30 patients in each group. Then the success rate of glottis exposure, the operative time, success rate and complication rate of tracheal intubation were compared between the two groups. Subgroup assessment between the junior emergency resident doctor (A group) and the senior emergency resident doctor (B group) was conducted.Results:According to Cormack Lehan grades, the success rate of glottis exposure in VL group was higher than that in TL group ( P>0.05), and the success rate of Grade I in VL group was significant higer than that in TL group: 56.70%(17/30) vs. 30.00%(9/30), P<0.05. The trial times of successful intubation cases and the operative time of successful intubation cases were significantly less than those in TL group (1.30 vs 1.67, P = 0.049) and (56.37 s vs 67.12 s, P<0.05). In the subgroup, the one-time success rate of tracheal intubation in A-TL group was significantly lower than that in B-TL group (4/15 vs. 11/15, P<0.05), while the one-time success rate of tracheal intubation in A-VL group was 60.00%, which is lower than that in B-VL group ( P>0.05). The operative time consumed for successful intubation in A-TL group was significantly longer than that in B-TL group: 78.00 s vs. 55.57 s, P<0.05, while the operative time in A-VL group was a little longer than that in B-VL group ( P>0.05). Conclusions:Visual laryngoscope used in spontaneous respiration tracheal intubation can not only increase the success rate of glottis exposure, decrease trial times and shorten operative time of intubation, but also improve the success rate and decrease the complication rate of emergency tracheal intubation.

Objective To explore the CT and MRI imaging and clinicopathological features of extranodal NK/T cell lymphoma (NK/TCL). Methods Sixty-six patients with NK/TCL diagnosed from 2002 June to 2016 April in Beijing Tongren Hospital with intact CT and/or MRI imaging results were enrolled in this study. All the patients had tailed clinical information and follow-up. The imaging and clinicopathological features were analyzed retrospectively and their prognostic value on overall survival was analyzed. Results There were 49 males and 17 females with median age of 42 years. The median follow-up time was 18 months. The cases showed surrounding invasions including 10 cases (15.2 %) in soft palate, 5 cases (7.6 %) in hard palate, 2 cases(3.0 %) in tonsil, 8 cases(12.1 %) in upper lip, 13 cases(19.7 %) in maxillofacial soft tissue, 9 cases (13.6 %) in eyelid, 10 cases (15.2 %) in orbital, 3 cases (4.5 %) in maxilla, 6 cases (9.1 %) in pterygopalatine fossa,6 cases(9.1 %)in infratemporal fossa,3 cases(4.5 %)in skull base, 3 cases(4.5 %) in eyeball and 2 cases (3.0 %) in brain tissue. Kaplan-Meier survival analysis found that the 2-year overall survival rates of the patients with the involvement of hard palate, upper lip, maxillofacial soft tissue, eyelid, orbital, maxillary, eyeball and brain organizer were lower than those of the patients without the involvement of these sites(χ2values were 4.470,4.041,4.456,13.933,8.986,4.000,44.121,6.527,16.822,respectively, all P< 0.05). Further multivariate Cox regression analysis showed that maxilla and brain involvement were independent adverse factors (RR=34.717, 95 % CI 3.404-354.035, P=0.003; RR=37.545, 95 % CI 3.188-442.187, P= 0.004). Conclusions MRI and CT examinations are of great value in diagnosis and prognostic assessment of NK/TCL. Clinicians can make correct and timely diagnosis by comprehensive clinical, radiological and pathological features and can make a detailed clinical assessment to give patients appropriate treatment,thus improving the outcome of the NK/TCL patients.

OBJECTIVE： To study the in vitro inhibitory effects and antioxidant activity of different solvent extracts of Boehmeria nivea leaves against influenza A virus（H1N1）， and to expand the medicinal parts of B. nivea and develop natural antiviral and antioxidant drugs. METHODS： The leaves of B. nivea were extracted with 95％ ethanol. The ethanol extract was dissolved by water heating， and extracted with different solvents to obtain petroleum ether phase， trichloromethane phase， ethyl acetate phase， n-butanol phase and aqueous phase extracts of B. nivea leaves. The toxicity of aqueous extract of B. nivea leaves （50-400 μg/mL） on Madin-Darby canine kidney （MDCK） cell line was investigated. Using ribavirin as positive control， MDCK cells were attacked by influenza A virus（H1N1）. Western blotting assay was used to detect the expression of nucleoproteins （NP） in viral infected cells after treated with same concentrations of petroleum ether phase， trichloromethane phase， ethyl acetate phase， n-butanol phase and aqueous phase extracts of B. nivea leaves （100 μg/mL）， different concentrations of aqueous phase extract solution of B. nivea leaves （50， 100， 200， 400 μg/mL） and different concentrations of ribavirin solution （0.31， 0.63， 1.25 μg/mL）. Using vitamin C as a positive control， hydroxyl radical（·OH） scavenging test， DPPH radical scavenging test and reduction test were used to investigate in vitro antioxidant activity of the extracts. RESULTS： Aqueous phase extract of B. nivea leaves with concentration less than 400 μg/mL was nontoxic to MDCK cells. The petroleum ether phase， trichloromethane phase， ethyl acetate phase and aqueous phase extracts at 100 g/mL could significantly reduce the expression of NP protein in influenza A virus（H1N1） infected cells （P＜0.01）. Different concentrations （50-400 μg/mL） of aqueous extract could significantly reduce the protein expression of NP （P＜0.01） in concentration-dependent manner. The in vitro antioxidant activity of petroleum ether phase and ethyl acetate phase was similar to that of vitamin C. CONCLUSIONS： B. nivea leaves extract have better anti-influenza A virus（H1N1） effects in vitro， and the extracts of petroleum ether phase and ethyl acetate phase show good antioxidant activity in vitro.

Objective: To investigate the difference in the microstructure of gray matter nucleus in different movement subtypes of Parkinson’s disease (PD) by diffusion kurtosis imaging (DKI) technique, and to analyze the correlation with clinical manifestations. Methods: Ninety-seven patients with PD and 83 healthy controls performed conventional MRI sequence and DKI sequence scan. The PD patients were classified into gait disorder subtype (PIGD, n=57) and tremor dominant subtype (TD, n=40)subtypes according to motor symptoms. Fractional anisotropy (FA), mean diffusivity (MD), axial diffusivity (Da), radial diffusivity(Dr), mean kurtosis (MK), axial kurtosis (Ka) and radial kurtosis (Dr) maps and data were generated by software after processing. DKI was performed for all subjects and data was collected from different brain regions in both hemispheres, including red nucleus(RN), substantia nigra pars reticulate(SNr), substantia nigra pars compacta(SNc), putamen(PUT), globus pallidus(GP), head of caudate nucleus (CN)and thalamus(THA). Results: TD showed a higher MMSE score(P=0.019), but lower modified Hoehn-Yahr score than that in PIGD (P<0.001), there was no significant difference of age of onset, sex, limbs of onset or disease duration between two PD subgroups. Compared with healthy controls, both TD and PIGD showed down-regulated MD, Da and Dr and up-regulated Ka values(P<0.001); MK(0.83±0.26, 0.80±0.18) was increased in SNr both in TD and PIGD, while SNc, PUT and GP (0.84±0.20, 0.75±0.07, 0.81±0.14)were decreased only in TD (P=0.017, P=0.010, P=0.020, P<0.001, P=0.002). The Kr values of PUT and CN(0.71±0.17, 0.72±0.14) were reduced in PIGD, while CN(0.70±0.14) were reduced in TD respectively (P=0.002, P=0.031, P=0.007). The MK was lower in TD than that in PIGD (t=-2.214, P=0.029), and no significant difference was found in other grey matter nuclei between TD and PIGD(P>0.05). Moreover, there was no significant correlation between DKI value and disease duration, MMSE score or Hoehn-Yahr scale (P>0.05) in TD and PIGD. Conclusion There is heterogeneity of clinical symptoms between these two subgroups of PD. DKI can quantify the microstructural changes of grey matter nucleus in different type PD patient.

[Abstract] Objective: To investigate the effect of HMGB1 gene on the growth of human epithelial ovarian cancer xenografts in nude mice, and to lay a foundation for finding new targets for the treatment of ovarian cancer. Methods: Human epithelial ovarian cancer SKOV3 cells in logarithmic growth phase were selected to establish a human epithelial ovarian cancer xenograft model in nude mice. Nude mice with successful model establishment were randomly divided into control group and HMGB1-siRNA group. On the 7th, 9th, 11th, 14th, and 16th days after cell inoculation, the same amount of saline and HMGB1-siRNA were respectively injected into two groups of mice under the armpit.After 3 weeks, the nude mice were sacrificed by cervical dislocation, the tumor tissues were separated, and the volume of the tumor was measured. The apoptosis of transplanted tumor cells was detected by Tunnel staining. The expressions of HMGB1, STAT3 and p-STAT3 were detected by Western blotting. The expression of vascular endothelial growth factorA(VEGF-A) and microvascularization were detected by immunohistochemistry. Results: Compared with the control group, the growth of tumor volume slowed down in HMGB1 siRNA group, and on the 21st day, the tumor volume of HMGB1-siRNA group was significantly smaller than that of the control group (P<0.05). HMGB1-siRNA successfully knocked down the expression of HMGB1 mRNA in transplanted tumor tissue. The apoptosis rate of tissue cells in HMGB1-siRNA group was significantly increased ([34±8]% vs [6±2]%, P=0.04), and the expressions of HMGB1 and p-STAT3 were significantly reduced (P<0.05). The expression of VEGF-Aand the number of microvessels were significantly lower than those of the control group (both P<0.05). Conclusion: Knockdown of HMGB1 gene reduces the expression of VEGF-A and microvessel formation possibly by inhibiting the HMGB1/STAT3 signaling pathway, thereby promoting the apoptosis of tumor tissues and slowing the growth of xenografts.