[Objective]To explore the differentiation of bone marrow stromal cells(BMSCs) induced by OECs conditioned medium.[Method]The purified OECs were acquired by seleclive culture medium and cultivated in the polylysine coated plate.After 9～12 days culture, OECs conditioned medium was obtained by 2000 r/min configuration and then co-cultivated with purified BMSCs which were 3th generation.The differentiation of BMSCs were observed under inverted microscope and were identified by immumofluorescence method.[Result]The OECs conditioned medium had obviously effects on BMSCs' differentiation.BMSCs differentiation into neuron-like cells ratio was 55% and differentiation into astroctes cells ratio was 23%.[Conclusion]The conditioned medium which comes from OECs culture supernatant has obviously effects on BMSCs' differentiation into neuron-like cells.

[Objective] To develop a new type nerve tissue engineering scaffolds to repair the defect of peripheral nerve and observe the growth of bone marrow stromal cells cultured in vitro on the scaffolds.[Method]The biomaterial was made of I-collagen and gelatin by freeze-drying method,the alignment regularities of microscopic channels and there course directions were observed under the scanning electronic microscope.The size of the micropores and the factor of porosity were also measured.The bone marrow stromal cells were seeded on the collagen scaffolds and cultured for 5 days,then the growth of bone marrow stromal cells cultured in the scaffolds was observed.[Result]All the scaffolds were circular cylinder,the microscopic channels were arranged in parallel manners,and the pore sizes of the channels were uniform.The bone marrow stromal cells were seeded in the scaffolds successfully.[Conclusion]The developed nerve tissue engineering scaffolds have good structure and biocompatibility,which can be used in the repair of the nerve injuries.

Objective To evaluate the patient-controlled paravertebral block (PCPB) in optimizing the cellular immune function when used after radical resection of pulmonary carcinoma performed via video-assisted thoracoscope in patients.Methods Forty-one ASA physical status Ⅰ or Ⅱ patients of both sexes,aged 50-64 yr,with body mass index of 20-25 kg/m2,of TNM staging Ⅰ or Ⅱ,undergoing radical resection of pulmonary carcinoma performed via video-assisted thoracoscope,were randomly divided into 2 groups using a random number table:PCIA group (n =21) and PCPB group (n =20).PCIA solution contained sufentanil 2 μg/kg in 100 ml of normal saline.The PCIA pump was set up to deliver a 2 ml bolus dose with a 15-min lockout interval and background infusion at 2 ml/h.In PCPB group,the patients received paravertebral injection of 0.2％ ropivocaine 5 ml at T5 level on the affected side under ultrasound guidance at the end of operation,and then received PCPB.PCPB solution contained 0.75％ ropivacaine 67 ml in 250 ml of normal saline,and the pump was set up to deliver a 5 ml bolus dose,with a 15-min lockout interval and background infusion at 5 ml/h.VAS score was maintained ≤ 3,and analgesia lasted until 50 h after operation.Before induction of anesthesia (baseline),at end of operation,and at 1,3 and 5 days after operation,peripheral venous blood samples were collected to determine the levels of regulatory T cells,natural killer cells and natural killer T cells (by flow cytometry) and plasma concentrations of interleukin-10 and transforming growth factor-β (by ELISA).Results Compared with group PCIA,the level of regulatory T cells was significantly decreased,the levels of natural killer cells and natural killer T cells were increased,and the plasma concentrations of interleukin-10 and transforming growth factor-β were decreased at 1 and 3 days after operation,and no significant change was found in the rate of cellular immune function decline after operation in group PCPB.Conclusion PCPB provides no significant difference clinically in optimizing the cellular immune function when used after radical resection of pulmonary carcinoma performed via video-assisted thoracoscope in the patients.

Objective:To study the clinical effect of Maizhiling tablets combined with salmon calcitonin in the treatment of elderly femoral neck fracture after artificial femoral head replacement and the influencing factors of hip joint function 6 months after operation.Methods:Prospective research methods were used in this article.A total of 84 elderly patients with femoral neck fracture who underwent artificial femoral head replacement in the Affiliated Hospital of North China University of Science and Technology from January 2017 to June 2019 were selected and randomly divided into the treatment group (41 cases) and the control group (43 cases) by the random number table method.The treatment group was treated with Maizhiling tablet and salmon calcitonin nasal spray plus basic calcium; the control group was treated with basic calcium.Independent sample t test was used to compare the visual analogue score (VAS) on the 5th day after operation, the time of complete disappearance of congestion in thigh, and the time of first ambulation after operation between the two groups.The bone mineral density (BMD) of contralateral femoral neck and Harris score of hip joint function were compared between the two groups at 14 days, 3 months and 6 months after operation by repeated measurement variance analysis.Multivariate Logistic regression was used to analyze the influencing factors of hip joint function at 6 months after operation.Results:VAS score on the 5th day after surgery: (6.34±1.54) points in the treatment group and (7.02±1.50) points in the control group.Complete regression time of thigh congestion: (12.12±2.27) d in the treatment group and (13.88±2.58) d in the control group.The first postoperative activity time was (5.93±1.52) d in the treatment group and (7.84±1.05) d in the control group, and the differences of the above indexes between the two groups were statistically significant ( t value was 2.051, 3.313, 6.673, respectively, all P<0.05). The BMD values of the contralateral femoral neck in the treatment group were (0.543±0.123), (0.561±0.119), (0.606±0.105) g/cm 2 at 14 days, 3 months and 6 months after operation, and (0.530±0.092), (0.517±0.089), (0.526±0.090) g/cm 2 in the control group.The results of variance analysis of repeated measurements showed that Fintra-group=55.726, P<0.01; Finter-group=4.206, P<0.05; Finteraction=57.654, P<0.01.There were significant differences between 3 months, 6 months and 14 days after operation in the treatment group (all P<0.01), and between 3 months and 6 months after operation ( P<0.01). In the control group, there was significant difference between 14 days and 3 months ( P<0.01), and between 3 months and 6 months ( P<0.01); 6 months after operation, the difference was statistically significant ( P<0.01). The Harris scores of hip joint in the treatment group were (82.12±8.18), (85.49±6.61), (87.10±6.57) points and (78.91±5.75), (81.44±6.42), (83.67±6.97) points in the control group.The results of repeated measurement ANOVA showed that Fintra-group=64.526, P<0.01; Finter-group=6.529, P<0.05; Finteraction=0.501, P>0.05.There were significant differences between 3 months, 6 months and 14 days after operation (all P<0.01). There were significant differences between 3 months and 6 months after operation ( P<0.05). There were significant differences between the groups 14 days, 3 months and 6 months after operation ( P<0.05 or P<0.01). Multivariate logistic regression analysis showed that age( OR=1.318, 95% CI1.002-1.732, P=0.048) and treatment method ( OR=29.168, 95% CI 1.030-824.623, P=0.048) were risk factors for hip function on the replacement side, and Harris score of hip function 14 days after surgery ( OR=0.624, 95% CI 0.447-0.873, P=0.006) were protective factors for hip function 6 months after surgery. Conclusion:Maizhiling tablets and salmon calcitonin nasal spray have good effects on removing blood stasis, relieving pain, effectively improving bone mineral density and promoting the recovery of joint function after artificial femoral head replacement for senile femoral neck fracture.The younger age of patients, postoperative treatment with Maizhiling tablets combined with salmon calcitonin and good recovery of hip function in 14 days after operation are the protective factors of good hip function in 6 months after operation.

Objective To investigate the dynamic changes of interleukin-1 receptor-associated kinases (IRAK-4) in hypoxic neurons and explore their role in regulation of inflammatory reaction. Methods The B35 cells exposed to hypoxia of 3% O2,5% CO2 and 92% N2 were cultured for 1,3,6, 12,24,48,72 and 96 hours respectively. Then, mRNA and protein expressions of IRAK-4 were detected by RT-PCR and Western blot, the expression of IRAK-4 in the cells were observed by laser scanning con-focal microscope (LSCM), and the concentration of IL-6 was measured by ELISA method. Results After hypoxia, the mRNA and protein expressions of IRAK-4 were increased at one hour, reached the peak at six hours (P<0.05), kept at a high level at 12 hours (P<0.05) , but decreased gradually to the normal oxygen level at 24 hours (P < 0.05) and to below the normal oxygen level at 48 hours (P < 0.05). Immunofluorescence results showed that the fluorescence intensity of IRAK-4 was gradually increased with time. The changes of IL-6 in the supernatant were positively correlated with protein expression of IRAK-4 (r =0.84, P < 0.05 ). Conclusions Hypoxia can increase the expression of IRAK-4 at transcription and translation levels in a certain period of time, which may participate in down-stream inflammatory reaction and lead to increase of IL-6 expression.

Objective:To investigate the risk factors of lower extremity venous thrombosis in elderly patients with femoral neck fracture during perioperative period after artificial bipolar femoral head replacement and the effect of Mailuoshutong in clinical prevention and treatment of thrombosis.Methods:A retrospective case-control study was conducted on the clinical data of 92 elderly patients with femoral neck fracture who were admitted to the Affiliated Hospital of North China University of Technology from January 2015 to January 2020.According to the treatment method, the patients were divided into an observation group (44 cases) and a control group (48 cases). Patients in the observation group were treated with the combination of Mailuoshutong Pill and low molecular weight heparin calcium.The control group was treated with low molecular weight heparin calcium.The patients in both groups started anti-coagulation therapy immediately after admission, and they stopped the drug one day before operation and continued to take the drug on the second day after operation.Venous ultrasound of both lower limbs was recorded at the time of admission, on the 7th day after admission, and 14 days after surgery.The changes in hemoglobin, red blood cell count and drainage volume before and after operation were recorded.Results:The incidence of thrombosis was 2 cases (4.54%) on the 7th day after admission in the observation group and 9 cases (18.75%) in the control group.The difference was statistically significant (χ 2=4.400, P=0.036). The incidence of thrombosis was 3 cases (6.82%) in the observation group and 11 cases (22.92%) in the control group 14 days after operation.The difference was statistically significant(χ 2=4.611, P=0.032). The hemoglobin changes of observation group and control group were (23.73±6.89) g/L and (22.10±5.18) g/L respectively on the first day before operation and 48 hours after operation.The red blood cell count changes were (0.67±0.32) × 1012/L and (0.56±0.36) × 1012/L respectively, and the drainage volume of drainage tube after operation was (100.27±23.73) ml and (102.40±20.90) ml, respectively.There was no significant difference in the above indexes between the two groups (all P>0.05). Multivariate Logistic regression analysis showed that only low molecular weight heparin calcium was used to prevent and treat thrombosis ( OR=10.281, 95% CI: 1.609-65.689, P=0.014); the elderly patients ( OR=1.190, 95% CI: 1.061-1.336, P=0.003) and the thrombosis at the time of admission ( OR=8.346, 95% CI: 1.773-39.281, P=0.007) were the risk factors for lower extremity venous thrombosis on the 14th day after surgery. Conclusion:Mailuoshutong pill combined with low molecular weight heparin calcium can safely and effectively treat lower extremity venous thrombosis in perioperative period of artificial femoral head replacement for femoral neck fracture in elderly patients.Mailuoshutong pill combined with low molecular weight heparin calcium was a protective factor for lower extremity venous thrombosis 14 d after operation, while advanced age and thrombosis at the time of admission were risk factors.

Objective:To report the clinical characteristics, diagnosis, and treatment of 2 cases of X-linked acrogigantism(X-LAG).Methods:The clinical information of two patients were retrospectively reported, and peripheral blood DNA was collected for copy number variations detection.Results:Both patients had onset at age of two, with common clinical characteristics including linear growth acceleration, mild facial coarsening, enlargement of hands and feet, increased appetite, and snoring, etc. The heights Z scores of the two patients before treatment were + 6.86 and + 6.53, respectively. Growth hormone(GH) glucose inhibition test showed that GH nadir values were over 1 ng/mL and insulin-like growth factor-Ⅰ(IGF-Ⅰ) were 586.0 ng/mL and 1 042.0 ng/mL, respectively. Patient 1 received three cycles of octreotide microspheres therapy followed by surgery, and achieved clinical and biochemical remission. Patient 2 had lanreotide for 5.5 years but failed biochemical remission. Microduplication of Xq26.3, which contained pathogenic gene G-protein coupled receptor 101(GPR101), was found in germline DNA of two patients through copy number variation detection, leading to the diagnosis of X-LAG.Conclusion:It should be cautious of X-LAG when children below 2 years old presents symptoms such as overgrowth and so on. Medication combined with surgery is effective.

Objective:Cushing′s disease(CD) is caused by the pituitary adrenocorticotroph hormone(ACTH) secreting adenomas, leading to increased serum cortisol levels and various abnormal metabolic processes. Untreated CD is linked to high mortality, thus it is critical to elucidate its pathogenesis. This study aims to explore the pathogenesis of pituitary ACTH adenomas using whole-genome sequencing analysis.Methods:Fresh tumor tissues and peripheral blood samples were collected in 9 confirmed cases of pituitary ACTH adenomas who underwent surgery. Whole genome sequencing was then performed, followed by analysis and verification of single nucleotide mutations, copy number variation(CNV) and chromosome structure variations.Results:Somatic USP8 mutations(p.Ser718del, p. Ser718Pro, p. Pro720Arg, p. Pro720Gln) were found in 5 patients, with a rate of 55.6%; CNV of USP8 was detected in 1 patient; TP53(p.Cys135Tyr), NF1(p.Val1049Glufs*11) and KMT2C(c.3323+ 1G>A) mutations were identified in 1 patient harboring wild-type USP8. CNV analysis showed a loss of heterozygosity in multiple chromosomes in a wild-type USP8 patient. Structural variations were found in 2 with unknown significance. No germline gene mutations were detected in this study.Conclusion:Somatic USP8 mutations, increased copy number of USP8, variations of tumor-related genes such as TP53 and extensive somatic CNV all contribute to pathogenesis of CD. Chromosomal structure variations may suggest high-risk pituitary ACTH adenomas, and call for frequent follow-up and aggressive treatment.

OBJECTIVE To establish the fingerprint of Zhuang medicine Jinmu granules and carry out chemometric analysis ， and determine the contents of three components . METHODS High performance liquid chromatography （HPLC） method was adopted. Using rutin as the reference ，HPLC fingerprints of 10 batches of Jinmu granules were drawn and similarity evaluation was performed by Similarity Evaluation of Chromatographic Fingerprint of Traditional Chinese Medicine （2012 edition）；the common peak was determined by comparing with mixed control ；SPSS 21.0 software was used for cluster analysis ，and SIMCA 14.1 software was used for principal component analysis and orthogonal partial least squares-discriminant analysis. The differential components affecting the quality of Jinmu granules were screened by taking the variable importance in projection （VIP）value ＞1 as the standard ；the HPLC method was used to determine the contents of astilbin ，polydatin and berberine hydrochloride in Jinmu granules. RESULTS There were 22 common peaks in 10 batches of Jinmu granules ，and the similarities were 0.962-0.997；five common peaks were identified ，namely gallic acid （peak 2），polydatin（peak 9），rutin（peak 11），astilbin（peak 13）and kaempferol （peak 20）. The results of cluster analysis showed that 10 batches of Jinmu granules could be clustered into 3 categories：S1 and S 3-S4 were grouped into one category ；S5-S6 and S 9 were grouped into one category ；S2，S7-S8 and S 10 were grouped into one category. The results of principal component analysis showed that the parameter of model interpretation was 0.951 and that of prediction ability was 0.723. The classification results were basically consistent with cluster analysis. The classifica tion results of orthogonal partial least squares- com discriminant analysis were also ba sically consistent with clus- ter analysis. The common peaks with VIP value ＞1 in the order were peak 7＞peak 11（rutin）＞peak 17＞peak 13（astilbin）＞ peak 3＞peak 8＞peak 6＞peak 16 respectively. The linear ranges of astilbin ，polydatin and berberine hydrochloride were 0.012 6- 1.225 0，0.010 8-1.052 5 and 0.020 0-1.562 5 mg/mL，respectively（all R 2＝0.999 9）. RSDs of precision ，stability（24 h）and repeatability tests were all less than 3％. The average recoveries were 99.48％（RSD＝2.67％，n＝9），98.57％（RSD＝1.77％，n＝ 9）and 100.84％（RSD＝2.49％，n＝9）. The contents were 1.221 0-7.011 6，2.251 1-4.462 9，1.252 4-3.328 7 mg/g，respectively. CONCLUSIONS Established fingerprint and the method of content determination are accurate ，stable and simple. Combined with chemometric analysis ，it can be used for the quality control and evaluation of Zhuang medicine Jinmu granules.