Objective To observe the effect of Astragalus injection on the expressions of inflammatory cytokines in human primary macrophages stimulated by lipoteichoic acid (LTA) and lipopolysaccharide (LPS),and investigate its effects on inflammatory reactions of Gram-positive (G+) and Gram-negative (G-) bacteria sepsis and its mechanisms.Methods Percoll density gradient centrifugation was used to isolate the human peripheral blood mononuclear cells,then they were purified by immune Anti-Biotin Microbeads with magnetic character and under the induction of recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF),the cells were cultivated for 12 days in vitro,eventually the human monocyte-derived macrophage was formed.The cultured human macrophages were inoculated in 96-well plates (each group 3 wells) and 6-well plates (each group 3 wells).The cells were divided into control group (200 μL DMEM added in each well),LTA 1 mg/L group,LPS 0.1 mg/L group and low astragalus injection (0.1 mg/L) and high astragalus injection (0.2 mg/L) dose groups.After the incubator plates were put in an incubator for 24 hours,the protein content of IL-8 and IL-10 in supernatant were detected by enzymelinked immunosorbent assay (ELISA),and the mRNA expression levels of IL-8 and IL-10 were detected by real time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-qPCR).Results LTA and LPS all can obviously up-regulate the expression levels of pro-inflammatory factor IL-8 and anti-inflammatory factor IL-10 of macrophage.The expressions of IL-8 and IL-10 protein and mRNA in LTA group and LPS group were significantly higher than those in control group after cuhure for 8 hours and 24 hours,the degrees of increment were more significantly at 24 hours [LTA stimute group:IL-8 protein (ng/L,× 103):41.57± 1.90 vs.1.58 ±0.24,IL-8 mRNA (A value):21.49±8.05 vs.1.00±0.16;IL-10 protein (ng/L):5.90±3.02 vs.2.91 ± 1.54,IL-10 mRNA (A value):1.35±0.34 vs.0.95±0.14;LPS stimute group:IL-8 protein (ng/L,× 103):345.00±22.80 vs.5.60±0.31,IL-8 mRNA (A value):29.84 ± 8.93 vs.1.00 ± 0.16,IL-10 protein (ng/L):122.37 ± 39.26 vs.44.79 ± 3.67,IL-10 mRNA (A value):7.38 ± 1.58 vs.1.35 ± 0.34,all P ＜ 0.05].The Astragalus injection could regulate LTA and LPS to stimulate the macrophage to decrease the expression levels of pro-inflammatory factor IL-8 protein and mRNA and increase the expression levels of anti-inflammatory factor IL-10 protein and mRNA in the macrophage;the changes of regulatory effect in the 24 hour-culture of Astragalus injection high dose group was the most significant [LTA stimute group:IL-8 protein (ng/L,×103):22.63±1.91 vs.41.57±1.90,IL-8 mRNA (A value):12.10±1.93 vs.21.49±8.05,IL-10 protein (ng/L):14.03±2.22 vs.5.90±3.02,IL-10 mRNA (A value):10.37±6.08 vs.1.35±0.34;LPS stimute group:IL-8 protein (ng/L,× 103):167.75 ± 19.90 vs.345.01 ±22.80,IL-8 mRNA (A value):15.61 ± 3.63 vs.29.84±8.93;IL-10 protein (ng/L):243.22±14.41 vs.122.37±39.26,IL-10 mRNA (A value):16.14±4.10 vs.7.38± 1.58,all P ＜ 0.05].Conclusions In the process of inflammatory response,the pro-inflammatory and anti-inflammatory factors co-exist simultaneously.Astragalus injection can inhibit the expression levels of pro-inflammatory factor gene and protein in the inflammatory response of G+ and G-bacteria sepsis and in the mean time,it can promote the expression levels of anti-inflammatory factor gene and protein,thus the immune mechanism of sepsis is affected,achieving the balance between pro-inflammation and anti-inflammation.

Objective To explore the clinico-pathological features, immunophenotype, treatment and prognosis of urologic primary primitive neuroectodermal tumor (PNET). Methods The clinical data of 3 patients with urologic PNET were analyzed retrospectively. All patients were male, aged 29, 32 and 75 years respectively. Two of the lesions were located in the kidney, and the third was located in the bladder. The sizes of renal tumors were 7.7 cm×6.2 cm and 12.6 cm×9.4 cm respectively. Imaging examinations revealed a well-defined mass with inhomogeneous echo inside. The size of bladder tumor was 10.0 cm×10.0 cm. CT scan demonstrated irregular thickening of the bladder wall, and the density of the wall was inhomogeneous. In the 2 cases of renal PNET radical surgery was performed, while an emergency palliative surgery to remove a blood clot and biopsy were performed in the bladder PNET case. Results In light microscope, the tumors were characterized by uniform small round or oval cells and nest-like or dense sheet structures surrounded by sparse fibrovascular stroma. Homer-Wright rosettes or pseudorosettes were observed, as well as mitoses. Immunohistochemical study revealed that all cases showed positive staining for CD99, synaptophysin and vimentin. One of the renal tumor cells showed positive for CD56, and the other renal tumor and urocystic tumor cells were focally positive for chromogranin A. Additionally, in 1 of the cases of renal tumor there was a high positive rate of 80% for Ki67 staining while the other case showed less than 5%. All 3 cases were eventually diagnosed as PNET. The first renal tumor case was not treated with radiotherapy and chemotherapy postoperatively, and the patient died of recurrence 14 months after surgery. Both the second renal tumor case and the bladder tumor case underwent chemotherapy postoperatively, and they died 4 and 6 months after surgery respectively. Conclusions The urologic primary PNET is a very rare, highly malignant soft tissue tumor, and the diagnosis must be based on pathologic findings and immunohistochemical phenotypes. The multimodal treatment for urologic primary PNET consists of surgery, chemotherapy and radiotherapy.

Objective To analyze and summarize the clinical-pathological features,immunophenotype and prognosis of the lymphoepithelioma-like gastric carcinoma (LELGC).Methods The clinical,radiographic and histological data of four patients with LELGC were retrospectively analyzed.The expression of cytokeratin (CK),CD20,CD3,CD4,CD8,E-cadherin,β-catenin,bcl-2,p16,p53,p63,c-erbB-2,cyclin D1,Ki67 and DNA methyl-transferase 1 (DNMT1) in tumor was detected by Epstein-Barr virus-encoded small RNA (EBER) in situ hybridization and immunohistochemical methods.Results The size of four tumor was 1.8 cm× 1.6 cm,1.5 cm× 2.0 cm,2.5 cm× 2.0 cm and 4.0 cm× 2.5 cm.Under light microscope,tumor cells appeared like cords,small lumps or scattered single infiltration with vacuolized nucleus; clear nucleoli and obvious interstitial lymphocytic infiltration.The results of immunohistochemical examination indicated that in four tumors CK was positive in membrane of all the tumor cells,while EBER was positive in all cell nucleus.The number of lymphocytes with CD3 positive was over those with CD20 positive,which was mainly CD8 positive lymphocytes.The percentage of E cadherin and β-catenin positive in the cell membrane of four tumors was between 10 ％ and 90 ％,and two cases with β-catenin positive in cytoplasm.The expressions of DNMT1,cyclin D1 and bcl-2 were all positive,while p16 and c-erbB-2 were all negative.The expression of p63 was positive in only one case,and p53 was negative in one case.The percentage of Ki67 positive was 40％,15％,60％ and 40％,respectively.Conclusions LELGC is a rare neoplasm with better prognosis.The features of clinical pathology and immunohistology may help to make a correct diagnosis.

Background and purpose:Δ133p53 can promote tumor cell growth, but the exact mechanism is not clear. This study was aimed to observe the expression and signiifcant of the p53 isoformsΔ133p53 and p53 gene downstream molecules MDM2 and cyclin G1 genes by 5-FU-MKN45 gastric cancer cell line model. Methods:Af-ter using different concentrations of 5-FU (50μg/mL, 100μg/mL) to human gastric cancer cell line MKN45, inhibition rate should be detected by MTT assay, the changes ofΔ133p53 mRNA, MDM2 mRNA and cyclin G1 mRNA express-ing were detected by reverse transcription-polymerase chain reaction (RT-PCR). Differences between these groups were analyzed by ANOVA, comparisons within groups were analyzed by t-test, bivariate correlation was analyzed by Pearson linear correlation. Results:MTT results showed that with the increased concentration of 5-FU and the extension of time, the cell inhibition rates increased gradually. The inhibition rates of 50μg/mL 5-FU were 41.10%, 54.79%and 68.48%, for culturing 24, 48 and 72 hours. There were statistically signiifcant differences between the groups(F=45.52, P=0.00). The inhibition rates of 100μg/mL 5-FU were 69.53%, 78.21%and 86.92%, for culturing 24, 48 and 72 hours. There were statistically signiifcant differences between the groups(F=85.58,P=0.00). The inhibition rates of 50 and 100μg/mL were 41.10%and 69.53%, for culturing 24 hours. There were statistically signiifcant differences between the groups(F=51.29, P=0.00). The inhibition rates of 50 and 100μg/mL were 54.79%and 78.21%, for culturing 48 hours. There were statistically signiifcant differences between the groups(F=51.29, P=0.00). The inhibition rates of 50 and 100μg/mL were 68.48%and 86.82%, for culturing 72 hours. There were statistically signiifcant differences between the groups(104.91, P=0.00). RT-PCR results showed that with the increase of the concentration of 5-FU, theΔ133p53 mRNA, MDM2 mRNA and cyclin G1 mRNA expression gradually declined in gastric cancer cell line MKN45 cells, and there were statistically signiifcant differences between the groups(F=738.532, 1 396.607, 2 785.56,P=0.00). Cor-relation analysis showed that the expressions ofΔ133p53 mRNA and MDM2 mRNA in gastric cancer were positively correlated (r=0.871, P=0.01), while the expression of cyclin G1 mRNA and p53 mRNA had no obvious relevance (P=0.13). Conclusion:In the 5-FU-MKN45 gastric cancer cell line model, anti-tumor pathway ofΔ133p53 isomers is related with MDM2 but was not related with cyclin G1.

Background and purpose: Little about the function of p53 isoforms in gastric cancer was reported. This study was designed to explore the role ofΔ133p53 in the effect of recombinant mutant human tumor necrosis factor (rmhTNF) on gastric cancer cells, and provide a new basis for the diagnostics and therapeutics of gastric carcinoma. Methods: MKN45 (withΔ133p53 expression) or SGC7901 (withoutΔ133p53 expression) cells were treated with rmhTNF of different concentrations only or combined with 5-FU (a traditional gastric cancer cellular killer), and the growth inhibition rate and apoptosis was detected by CCK-8 and lfow cytometry. mRNA expressions ofΔ133p53, Gadd45αand CyclinB1 were measured by nested reverse transcription-polymerase chain reaction (nRT-PCR) or real-time polymerase chain reaction(RT-PCR). Results:On MKN-45 cells with positiveΔ133p53 expression, the inhibitory effect of rmhTNF was signiifcant, the inhibition rates of 50 and 500 IU/mL rmhTNF were 24.82%, 72.33%after culturing for 24 h (t=-9.558, P<0.01);also, the inhibitory effect of 5-FU was improved by rmhTNF remarkably in time-and dose-dependence, the inhibition rates of 5-FU (25 μg/mL), rmhTNF (50 IU/mL) combined with 5-FU (25 μg/mL), rmhTNF (500 IU/mL) combined with 5-FU (25 μg/mL) were 18.20%, 48.66%, 59.83%, separately, after culturing for 24 h (F=82.742, P<0.01);the inhibition rates of rmhTNF (50 IU/mL) combined with 5-FU (25 μg/mL) were 48.66%, 68.20%, 85.23%, separately, after culturing for 24 h, 48 h and 72 h (F=128.583, P<0.01). However, on SGC-7901 cells with negativeΔ133p53 expression, no growth inhibition was showed by rmhTNF only, the inhibition rates of 50 and 500 IU/mL were 2.74%, 3.25%after culturing for 24 h (t=-0.121, P>0.05). In apoptosis test, the apopto-sis-enhancing effect of rmhTNF was signiifcant on MKN45 cells, and the apoptosis-enhancing effect of 5-FU was fur-ther promoted signiifcantly by rmhTNF, the apoptosis of rmhTNF (50 IU/mL), rmhTNF (50 IU/mL) combined with 5-FU (25 μg/mL), rmhTNF (500 IU/mL) combined with 5-FU (25 μg/mL) were 18.20%, 48.66%, 59.83%, separately, after culturing for 24 h (F=123.931, P<0.05). In mRNA measurement, down-regulation ofΔ133p53 and CyclinB1, up-regula-tion of Gadd45αwere signiifcant in MKN45 cells treated by rmhTNF alone or combined with 5-FU. In nRT-PCR anal-ysis, the mRNA levels ofΔ133p53 were relatively 0.886, 0.499, 0.330, 0.161 (F=240.927, P<0.01);In real-time PCR analysis, the mRNA levels of Gadd45αwere 1.227, 1.694, 3.394, and the mRNA levels of CyclinB1 were 1.221, 0.722, 0.316, relatively. The expression ofΔ133p53 was positively related to CyclinB1 (r=0.977, P<0.01), but negatively re-lated to Gadd45α(r=-0.950, P<0.01). Conclusion:InΔ133p53 positively expressed MKN45 cells, rmhTNF showed as an effective tumor inhibitor and an enhancer of 5-FU as well, and this effect might be helped by two p53 down-stream molecules CyclinB1 and Gadd45α. The results suggest thatΔ133p53 might be a key target for the biological effect of rmhTNF against gastric cancer.

Objective To investigate the learning outcome of clinical scenarios discussion im-plemented in teaching medical students before clinical probation. Methods Using standardized pa-tients (SP) to simulate the clinical common difficult situation, all eight year program clinical medicine students (84) of Grade 2008 who were about to enter the clinical probation were given clinical commu-nication education guidance. Through five classification variables questionnaire and return visit, the teaching effect was evaluate. Epidata 3.0 was used to input data, and the SPSS 15.0 was used to make descriptive analysis of the questionnaire and proportion comparison. Results We found 35.4%(29) and 63.4%(52) of subjects liked and approval this training, respectively;90.2%(74) of the subjects thought the diffi-culty of this train was moderate; 77.3% (65) of subjects believed that it was necessary to set the clinical scenarios discussion before clinical probation. Conclusions The implementation of clinical scenarios discussion before clinical probation was effective on inducing the medical students to pay attention to the doctor-patient communication, and deep thinking about the communication notice mat-ters and cop-ing styles in clinical environment.

Objective:To investigate the effects of early applying of basic fibroblast growth factor (bFGF) on corneal haze formation after surface ablation surgery in rabbits.Methods:The right eyes of 60 healthy New Zealand white rabbits received photorefractive keratectomy (PRK) and were randomized into PRK+ normal saline group, PRK+ bFGF group and simple PRK group, with 20 rabbits in each group.Normal saline solution and bFGF were topically administered according to grouping, respectively, 3 times per day, 1 drop for each time until the sacrifice of the animals, and no drug was used in the PRK group.Another 8 normal rabbits were served as blank control group.The corneal healing response and haze formation were evaluated by anterior segment photography and anterior segment optical coherence tomography (AS-OCT) and graded based on Fantes criteria.Corneal histopathology was examined by hematoxylin-eosin staining.Immunohistochemistry was used to detect the expression of transforming growth factor-β 1(TGF-β 1), α-smooth muscle actin (α-SMA) and matrix metalloproteinase-2 (MMP-2) in cornea.This study protocol was approved by the Experimental Animal Ethic Committee of Affiliated Hospital of Binzhou Medical University (20180209-03). The use and care of the animals complied with the Statement of ARVO. Results:The corneal epithelium was completely healed in 3-4 days following surgery and there was not significantly different in healing time among the three groups.( F=0.57, P=0.57). The haze grading was significantly different among different groups at different time points ( Fgroup=41.736, P<0.01; Ftime=129.445, P<0.01) and showed the highest score in the PRK+ bFGF group on the 28th day after operation.On the 7th day after surgery, AS-OCT image showed that the surface reflection of corneal epithelium was continuous and smooth and corneal epithelium was not tightly attached to the superficial stromal layer; the reflection of the superficial stromal layer was enhanced in all the operation groups.The proliferation of corneal epithelial cells and superficial stromal layer in the operation area were seen under the optical microscope, and the arrangement of collagen fibers in the stromal layer was disordered with the most obvious changes in the PRK+ bFGF group in comparison with the PRK+ normal saline group and the simple PRK group, and these findings became worse on postoperative 28 days.The corneal epithelial surface reflection in the blank control group was continuous and smooth.Immunohistochemistry showed that a few MMP-2 positive cells were seen in the blank control group.TGF-β 1, α-SMA and MMP-2 proteins were positively expressed in the corneas 7 days after surgery in the three groups, and their expressions were the most obvious in the PRK+ bFGF in comparison with the PRK+ normal saline group and the PRK group and were enhanced 28 days after operation, showing statistically differences (all at P<0.05). Conclusions:Early application of bFGF following surface ablation surgery promotes the proliferation of corneal epithelial cells and irregular arrangement of collagen in the superficial stromal layer, which is associated with the expressions of haze-related factors TGF-β 1, α-SMA and MMP-2 in corneas.

BACKGROUND:There are differentially expressed genes in acute intracerebral hemorrhage,which are related to the occurrence and development of intracerebral hemorrhage. OBJECTIVE:To screen differentially expressed genes and key genes in brain tissue of a rat model with acute intracerebral hemorrhage,to validate them through qPCR,and to analyze the relationships between key genes and the neurological function and brain tissue water content after intracerebral hemorrhage. METHODS:Seventy-eight Sprague-Dawley rats were randomly divided into two groups:in intracerebral hemorrhage group,a rat model of acute intracerebral hemorrhage was made using collagenase injection at the right caudate nucleus;and in sham-operated group,rats were injected with equal amount of saline at the same site.RNA was extracted from rat brain tissues of both groups using the TRIzol method and transcriptome sequencing technology was used to identify differentially expressed genes in brain tissues of acute intracerebral hemorrhage,which were then verified by qPCR and analyzed for the relationships between the genes and neurological function and brain tissue water content after intracerebral hemorrhage.And the key genes were analyzed by GO and KEGG functional enrichment analysis in combination with bioinformatics. RESULTS AND CONCLUSION:Ten key genes were identified,including CXCL8,SERPINE1,TFPI2,CXCR4,GDA,KCNQ5,ERICH3,SCN3B,CACNA1E,and CCL20.The contents of GDA,KCNQ5,ERICH3,SCN3B,and CACNA1E in the intracerebral hemorrhage group were lower than those in the sham-operated group(P＜0.05).The contents of CXCL8,SERPINE1,TFPI2,CXCR4 and CCL20 in the intracerebral hemorrhage group were higher than those in the sham-operated group(P＜0.05).The contents of GDA,KCNQ5,ERICH3,SCN3B,and CACNA1E were positively correlated with brain tissue water content and neurologic deficit score(P＜0.05),while the contents of CXCL8,SERPINE1,TFPI2,CXCR4 and CCL20 were negatively correlated with brain tissue water content and neurologic deficit score(P＜0.05).GO analysis indicated that differentially expressed genes were mainly enriched in two biological processes(leukocyte chemotaxis and chemokine-mediated signaling pathways),two cell components(cation channel complexes and ion channel complexes),and two molecular functions(gated channel activity and ion channel activity).KEGG analysis indicated that differentially expressed genes were concentrated in tumor necrosis factor signaling pathway,glutamatergic synapses and GABAergic synapses.To conclude,the differentially expressed genes in intracerebral hemorrhage include CXCL8,SERPINE1,TFPI2,CXCR4,GDA,KCNQ5,ERICH3,SCN3B,CACNA1E,and CCL20,and these genes are related to brain tissue water content and neurological function after intracerebral hemorrhage.These genes are mainly enriched in cell components,binding functions,cellular protrusions,and other related biological functions.

OBJECTIVE: To determine the prevalence and incidence of illness of two-week duration, and the factors influencing these, among residents 15 years and older in four counties of Hunan Province. METHODS: Data were sampled from four counties of Hunan Province for the Fourth National Health Service Survey. Incidence and two-week prevalence of disease were used to assess the health service needs of residents. A non-conditional, stepwise logistic regression was employed to explore the influencing factors. RESULTS: The two-week prevalence and incidence were 11.5% and 3.9%, respectively, in four counties of Hunan. The three leading diseases of two-week prevalence were: respiratory diseases, digestive diseases, and musculoskeletal diseases. Non-conditional stepwise logistic regression showed that urban residents had 0.64 times the risk of two-week illness compared with the rural residents (P< 0.05); residents in the 45-59 year age group and the 60+ year age group had 1.69 and 2.62 times the risk of two-week illness compared with residents in the 15-44 year age group, respectively (P<0.05). The widowed had 1.91 times the risk of prevalence of two-week illness contrasted to singles (P<0.05); the students had 0.29 times the risk of two-week illness contrasted to the workers (P<0.05); urban residents had 0.63 times the risk of two-week illness compared with the rural (P<0.05); the widowed had 2.37 times the risk of incidence of two-week illness compared with singles (P<0.05). CONCLUSION The majority of health service needs of residents of four counties is generated by three diseases: respiratory diseases, digestive diseases, and musculoskeletal diseases. Relatively, rural residents, the elderly, employed persons and the widowed have higher health service needs than others and deserve specific attention.
Adolescent ; Adult ; Aged ; China ; epidemiology ; Community Health Services ; statistics & numerical data ; Digestive System Diseases ; epidemiology ; Female ; Humans ; Incidence ; Male ; Middle Aged ; Musculoskeletal Diseases ; epidemiology ; Respiratory Tract Diseases ; epidemiology ; Sampling Studies ; Surveys and Questionnaires ; Young Adult

Objective:To compare the changes of serum calcium level before and after surgical resection in patients with primary hyperparathyroidism.Methods:Two hundred and seventy-one patients with primary hyperparathyroidism were enrolled from Dec 1992 to Dec 2020 in Beijing Jishuitan Hospital. Serum calcium concentrations were measured before operation, 20 min during surgery, then 2 weeks 1-6 months , 7-12 months and 1 year respectively after operation. The baseline data of postoperative serum calcium such as sex, age, other genetic endocrine diseases, osteopathia and urolithiasis were calculated. The generalized estimation equation was used to analyze the changes of serum calcium in different types of patients before and after operation.Results:The most common postoperative hypocalcemia occurred within 2 weeks, and it occurred frequently half a year after surgery. There was no significant difference in blood calcium between male patients ( t=0.875, P=1.000) and patients with bone lesions ( t=0.034, P=3.049) from 1 to 6 months after surgery and 2 weeks after surgery. Blood calcium level in patients aged 15-35 years old from 1 to 6 months ( t=0.239, P=1.000) , from 7 to 12 months ( t=1.380, P=0.935) and 2 weeks after surgery was not statistically different. The change of bone mineral density was correlated with the change of blood calcium after operation ( F=6.895, P=0.004). Conclusions:The incidence of hypocalcemia was the highest in patients with hyperparathyroidism 2 weeks after surgery, and the blood calcium level was stable within the normal range 1 year later. The blood calcium value of male patients was still at a lower level than that of female patients within six months after surgery. In patients with bone disease, the blood calcium value was lower and recovered slowly 2 weeks after surgery. The blood calcium value of patients aged 15-35 was at a low level within 1 year after surgery.