Objective To observe and compare the clinical effect of aerosol inhaled and systemic glucocorticoid treatment and its influence on the expression of procalcitonin ( PCT) and hyper-sensitive C reactive protein ( hs-CRP) in elderly patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD).Methods 146 elderly patients with AECOPD were enrolled prospectively.The patients were divided in three groups randomly, including inhalation group (48 cases referred as 2 mg of budesonide nebulization every 8 hours with therapeutic course of 5 days), intravenous group (50 cases referred as 40 mg methylprednisolone intravenous injection once daily with therapeutic course of 5 days), and control group (48 cases with no glucocorticoid ) .All the patients were treated with routine oxygen therapy, anti-inflammation, bronchodilators (β2 receptor agonist and doxofylline) and mucolytics, and so on.The levels of serum PCT and hs-CRP were detected before and after five days treatment, and also the clinical parameters such as symptoms, blood gas analysis, lung function, hospital days, fasting blood glucose and adverse reaction were simultaneously recorded. Results The general clinical data demonstrated no significant difference among the groups.Compared with the control group, the total effective rate, improvement rate of lung function (FEV1%pred and FEV1/FVC) and of blood gas parameters (pH value, PaO2 and PaCO2) in the inhalation group and intravenous group were higher, with shorter hospital days(P<0.05), while with no obvious difference between the two group.The levels of the serum PCT and hs-CRP were decreased in the patients enrolled after treatment, compared with the prior treatment (P<0.05).Compared with the control group, the levels of serum PCT and hs-CRP in the inhalation group and intravenous group decreased significantly, while with no obvious difference between the two groups.The levels of fasting blood glucose in the intravenous group were the highest;compared with the control group, the adverse effect incidence of the inhalation and intravenous groups were higher obviously, with the most obvious in the intravenous group ( P<0.05 ). Conclusion Application of glucocorticoid is benefit to improving the clinical symptom and lung function, and to rectifying hypoxemia and to shortening hospital days.The aerosol inhaled and systemic glucocorticoid application shows equivalent clinical efficiency on anti-airway inflammation.The glucocorticoid inhalation has fewer adverse effects and is recommended to treat the elderly patients with AECOPD .

Deflazacort (DFZ, a prodrug) is well absorbed and rapidly metabolized into the active metabolite 21-hydroxydeflazacort (21-OH DFZ) after oral administration. The aim of this study is to evaluate the pharmacokinetic properties of 21-OH DFZ in healthy Chinese volunteers after a single and multiple oral administration of DFZ tablets under fed condition. Twelve volunteers (six males and six females) were administered a single dose of 6 mg or 12 mg or 24 mg of DFZ in three different periods separately, according to the 3 x 3 Latin square design. Between each administration period there was a washout period of one week. The multiple-dose study of 12 mg dose DFZ per day for 7 consecutive days was started after a 1 w washout period when the single-dose study completed. The pharmacokinetic parameters of 21-OH DFZ after the single oral administration of 6 mg, 12 mg and 24 mg DFZ tablets were as follows: (37.7 +/- 11.6), (61.5 +/- 17.7) and (123 +/- 23) ng x mL(-1) for C(max); (1.90 +/- 0.32), (1.96 +/- 0.27) and (2.13 +/- 0.34) h for t1/2; (96.6 +/- 25.9), (190 +/- 44) and (422 +/- 107) ng x h x mL(-1) for AUC(0-14 h), respectively. After the multiple dose administration, the mean plasma concentration at steady-state C(av) was (7.00 +/- 1.66) ng x mL(-1) and the degree of plasma concentration fluctuation DF was 7.7 +/- 1.2. The results showed that the pharmacokinetic characteristics of 21-OH DFZ in healthy Chinese volunteers were linear over the dose range of 6 to 24 mg. No significant gender differences were found in the pharmacokinetics of 21-OH DFZ in healthy Chinese volunteers. After the multiple dose administration of 12 mg DFZ for 7 d, no accumulation of 21-OH DFZ in healthy Chinese volunteers was observed.

Objective To observe the clinical effects of Huoxue Dingxuan Capsules on treatment of vertebral artery type of cervical spondylosis, and evaluate its efficacy and safety. Methods Totally 130 cases of vertebral artery type of cervical spondylosis were randomly assigned to treatment group and control group, and treated with Huoxue Dingxuan Capsules and Sibelium capsules respectively. Integration method was used to compare the differences of main symptoms and signs, the overall efficacy and safety were evaluated, and relapse after six months was followed up. Results Both groups had a significant effect on the improvement of signs and symptoms in vertebral artery type of cervical spondylosis, the total efficiency was 94.03%(63/67) in treatment group and 82.54% (52/63) in control group. The treatment group had a better effect on the overall efficacy and relieving symptoms and signs compared with the control group (P<0.01). The mean blood flow velocity had a significant change after treatment (P<0.05), and the treatment group was better than the control group (P<0.01). Within six months, the treatment group had 4 cases (6.34%) with mild recurrence of symptoms and signs, and the control group had 18 cases (34.61%), the difference between the two groups was statistically significant (P<0.05), and no severe recurrence was found. The two groups showed no side effects. Conclusion Huoxue Dingxuan Capsules is safe and effective for treating vertebral artery type of cervical spondylosis, and the long-term efficacy and subjects’ compliance are better than oral Sibelium capsules.

Objective To explore the miRNA regulating the potential cancer-promoting gene CCL18 in cutaneous malignant melanoma.Methods Bioinformatics analysis was conducted by using online software miRanda and TargetScan,so as to predict the miRNA targeting CCL18 gene.Three kinds of C CL18 3'UTR dual-luciferase reporter vectors,including mutant 3'UTR vector (mutant 3'UTR group),wildtype 3'UTR vector (wild-type 3'UTR group) and empty vector (blank control group),as well as miRNA vectors carring selected miRNAs were constructed according to human gene sequence analysis,and then were used to co-transfect 293T cells.After 48-hour treatment,the cells were lysed for detection of luciferase activity.Real-time fluorescence-based quantitative PCR was performed to measure the expression of CCL 18 and selected miRNA in 14 fresh malignant melanoma tissue specimens and 14 paracancerous normal skin tissue specimens (control tissues),and their correlations were analyzed.Results Online software analysis showed that some miRNAs were identified to target the 3'UTR of CCL18 gene,including miR-183,miR-128 and miR-33a.Luciferase reporter vectors and miRNA vectors were constructed successfully.As luciferase activity assay showed,when miR-183 and miR-128 were bound to the CCL18 3'UTR,the luciferase activities were significantly higher in their mutant 3'UTR groups (11.63 ± 0.42;8.80 ± 0.49) than in their wild-type 3'UTR groups (4.86 ± 0.39;5.01 ± 0.54;both P ＜ 0.05) and blank control groups (2.41 ± 0.13;2.39 ± 0.05;both P ＜ 0.01),while there were no significant differences between miR-33a-hinding mutant 3'UTR group (6.41 ± 0.47) and miR-33a-binding wild-type 3'UTR group (6.16 ± 0.22,P ＞ 0.05).Real-time fluorescence-based quantitative PCR revealed higher mRNA expression of the CCL18 gene (3.52 ± 1.68),but lower expression of miR-183 (0.49 ± 0.32),miR-128 (0.30 ± 0.20) and miR-33a (0.46 ± 0.40) in the malignant melanoma tissues compared with the control tissues.The mRNA expression of the CCL18 gene was negatively correlated with the expression of miR-128 (rs =-0548,P ＜ 0.05),but showed no significant correlations with the expression of miR-183 and miR-33a (both P ＞ 0.05).Conclusion miR-128 may play a role in regulating the potential malignant melanoma-promoting gene CCL18.

Objective To study the mechanism of glutamine dipeptide on wound healing after operations on bums. Methods Totally, 30 burned patients were randomly divided into study group and control group (15 cases in each group). All patients received parental nutrition support after operation. Study group patients were supplemented with glutamine dipeptide at a dose of 0. 5 g · kg±1 · d±1. The plasma free hydroxyproline level was measured by a standard amino acid analyzer 1 day before operation and 7 days after operation and the wound healing time was recor± ded. Results The plasma free hydroxyproline levels of both groups were higher than the normal values before opera± tion but without significant difference [control group: (2. 24 ±0. 84) fig/ml, study group: (2. 32 ±0.92) μg/ml, normal value: (1.27 ±0.44) μg/ml]. On the post±operative 7 day, the plasma free hydroxyproline level of study group [ (4. 31 ±1. 05) μg/ml] was significantly higher than that of control group [ (3. 04± 1. 01) μ/ml] (P = 0.002). The wound healing time of study group [ (29. 7±5.3) d] was shorter than that of control group but with± out significant difference [ (33. 3 ±7.5) d, P = 0. 14 ]. Conclusion The intravenous supplementation of glutamine dipeptide may increase the plasma hydroxyproline level after operation and thus promote wound healing.

Objective: To explore the application of procalcitonin (PCT) and C-reactive protein(CRP) detection in acute exacerbation of chronic obstructive pulmonary disease(AECOPD) and its guiding value in the use of antimicrobial agents. Methods: From June 2014 to June 2018, 82 patients with AECOPD treated in the Third People's Hospital of Yuyao were selected.According to the difference of PCT and CRP levels at admission, the patients were divided into three groups: A, B and C group.A group (n=17): the serum PCT level was less than 0.10μg/L, treated with phlegm and asthma relief and symptomatic therapy without antibiotic therapy.B group (n=33): the serum PCT level was 0.10-0.25 μg/L, B group was not treated with antibiotics, if the level of PCT≥0.25 μg/L on the next day or the amplitude of increase≥30%, the treatment method was the same as that in C group.C group (n=32): the serum PCT level was more than 0.25 μg/L, C group was given expectoration and asthma relief, symptomatic treatment and antibiotics.The clinical data, laboratory test indicators and clinical efficacy of the three groups were compared. Results: There were no statistically significant differences in the general clinical data of the three groups (all P>0.05). The incidence of fever in the three groups were 2 cases (11.76%), 16 cases (50.00%) and 19 cases (57.58%), respectively; expiratory dyspnea in the three groups were 6 cases (35.29%), 25 cases (78.13%), 31 cases (93.94%), respectively; wheezing rale in the three groups were 4 cases (23.53%), 26 cases (81.25%), 33 cases (100.00%), respectively; wet sound in the three groups were 12 cases (70.59%), 27 cases (84.37%), 33 cases (100.00%), respectively, the differences of the above indicators among the three groups were statistically significant(χ²=15.827, 24.361, 30.284, 18.644, all P<0.05). The percentages of sputum positive patients in the three groups were 4 cases (23.53%), 10 cases (31.25%) and 23 cases (69.69%), respectively; the percentages of sputum culture positive patients were 5 cases (29.41%), 24 cases (75.00%), 28 cases (84.85%), respectively, the differences of the above indicators among the three groups were statistically significant(χ²=16.871, 24.644, all P<0.05). The WBC counts of the three groups were (4.27±1.92)×10⁹/L, (8.64±3.77)×10⁹/L, (18.06±4.87)×10⁹/L, respectively; the ratios of neutrophils were (54.12±3.48), (82.19±5.67), (90.07±9.33) , respectively; the levels of PCT of the three groups were (0.09±0.08) μg/L, (0.21±0.12) μg/L, (0.74±0.33) μg/L, respectively, there were significant differences in WBC count, ratio of neutrophils and PCT among the three groups (F=14.827, 25.825, 19.873, all P<0.05). The level of CRP among the three groups had no statistically significant difference (P>0.05). Patients in A group were not treated with antimicrobial agents and their clinical symptoms were improved.Fourteen patients without antimicrobial agents and 18 patients with improved clinical symptoms were treated with antimicrobial agents in B group.In C group, 20 patients with symptoms improved after treatment with antimicrobial agents, 12 patients with replacement of antimicrobial treatment regimen and 1 patient died. Conclusion PCT and CRP can be used as an index to evaluate the severity of AECOPD and a reference tool for the use or replacement of antimicrobial agents.

Objective To discuss the clinical value of sonographic patterns of thyroid nodules in 2015 American Thyroid Association (ATA) management guidelines for adult patients with thyroid nodules and differentiated thyroid cancer.Methods From January 2008 to December 2010,483 thyroid nodules in 480 patients who underwent ultrasound-guided fine-needle aspiration biopsy (US-FNAB) in Peking Union Medical College Hospital were included in this study.Ultrasound images of the thyroid nodules were reviewed and their sonographic patterns were determined as high suspicion,intermediate suspicion,low suspicion and very low suspicion according to 2015 ATA guidelines.112 nodules had surgical pathology.Diagnosis of other nodules depended on both US-FNAB pathology and follow-up of patients.Independent-sample t test was used to compare ages and sizes between benign and malignant nodules.Independent sample rank sum test was used to compare the malignancy risks between nodules of male and female patients,and between nodules that were solid,hypoechoic,with irregular margins,with microcalcifications and with taller than wide shape and each control group.Independent sample rank sum test was also used to compare malignancy risks of nodules with different sonographic pattems in 2015 ATA guidelines.Defining high-suspicion as positive,and intermediate to very low suspicion as negative,the diagnostic performance of sonographic patterns in 2015 ATA guidelines was calculated,including sensitivity,specificity,positive predictive value,negative predictive value,and accuracy.Results Of the 483 thyroid nodules,381 (78.9％) were benign and 102 (21.1％) were malignant proven by operation and follow-up.The malignancy rates were higher in nodules that were solid,hypoechoic,with irregular margins,with microcalcifications and with taller than wide shape than each control group,all of which had statistically significant differences (z=-6.255,-6.893,-13.000,-11.080,-6.718,P ＜ 0.001).Actual malignancy rates of nodules determined as high,intermediate,low and very low suspicion according to the ATA guidelines were 53.3％ (90/169),5.5％ (6/109),3.9％ (6/154),and 0 (0/51),respectively,with statistically significant differences between the four patterns (x2=161.462,P ＜ 0.001).When defining high suspicion as positive,and intermediate to very low suspicion as negative,the negative predictive value of sonographic patterns in ATA guidelines was relatively high (96.2％).Conclusion Sonographic patterns in 2015 ATA guidelines provide effective malignancy risk stratification for thyroid nodules.High suspicion is a good indication of US-FNAB for thyroid nodules and has relatively high negative predictive value.

The prescription of Xiangsu Powder （香苏散） reflects the exquisite balance in treating both the superficial and deep layers， harmonizing the body's vital energy and blood， and addressing the functions of the triple energizer simultaneously. Its application in treating cold epidemics has been a long time， yet its importance in modern disease treatment is often overlooked. Based on the treatment principles demonstrated by Xiangsu Powder in treating cold epidemics， it is proposed that in the treatment of modern epidemic diseases， the dual principles of cold and heat should not be neglected. When cold pathogens stagnate at the surface， they should be dispersed and detoxified， using methods that induce sweating to expel them from the surface. When cold and dampness coexist， efforts should be made to regulate the flow of vital energy， blood， and body fluids to restore the proper functioning of the three burners. When cold-induced pathogenic factors are severe and toxic， aromatic herbs should be utilized to dispel turbidity and avoid contamination. Throughout the entire course of treating epidemic diseases， attention should be paid to inducing sweating without damaging the body's vital energy， while also protecting the body's righteous qi， with the aim of providing refe-rence for the treatment of modern epidemic diseases.

Objective:To investigate the efficacy and safety of acute stent implantation during endovascular treatment for patients with emergent large vessel occlusion due to intracranial atherosclerotic stenosis.Methods:A retrospective analysis was carried out on 46 patients with emergent large vessel occlusion due to intracranial atherosclerotic stenosis who received endovascular treatment at the Strategic Support Force Medical Center from January 2015 to August 2022. Twenty-seven patients underwent balloon angioplasty alone and 19 patients underwent acute stent implantation. The baseline characteristics, modified thrombolysis in cerebral infarction (mTICI) score of the responsible vessels, modified Rankin scale (mRS) score 90 days after operation, incidence of symptomatic intracranial hemorrhage and mortality of the two groups were evaluated.Results:The proportion of effective recanalization of the offending vessels (mTICI≥2b) in the acute stenting group was slightly higher than that in the balloon angioplasty group (16/19 vs. 81.5%), but the difference was not statistically significant ( P>0.05). Besides, there was no significant difference in the median of mRS between the acute stenting group [3.0(0, 4.0)] and the balloon angioplasty group [4.0(1.0, 5.0)] 90 days after operation ( P>0.05). In terms of safety, the incidence of symptomatic intracranial hemorrhage and mortality were comparable between the two groups ( P>0.05). Conclusions:The effect of acute stent implantation during endovascular treatment for patients with emergent large vessel occlusion due to intracranial atherosclerotic stenosis is not inferior to that of balloon angioplasty, and it does not increase the risk of intracranial bleeding complications.

Objective:To investigate the influence of glucose regulatory protein 78 (GRP78) on prognosis and tumor cell proliferation of hepatocellular carcinoma.Methods:The experimental study and retrospective cohort study were conducted. Based on hepatocellular carcinoma tissue chip, in vitro culture of Huh7 and Hep3B hepatoma cells and LO2 normal hepatic cell, and combined with immunohistochemical staining, cell transfection, quantitative real-time polymerase chain reaction (qRT-PCR), Western blot detection, cell proliferation experiments, cell clone formation experiments and high-throughput transcription histological analysis, the GRP78 expression in hepatoma cells was analyzed. Huh7 and Hep3B hepatoma cells being transfected with the GRP78 gene-specific shRNA lentiviruses or the negative control shRNA lentivirus were set as the GRP78 gene-specific shRNA lentivirus group and the negative control shRNA lentivirus group respectively. Observation indicators: (1) GRP78 expression in hepatocellular carcinoma tissue and adjacent tissue and its correlation with the clinicopathological characteristics of hepatocellular carcinoma patients; (2) analysis of factors affecting the prognosis of hepatocellular carcinoma patients; (3) effects of inhibiting of GRP78 expression on the proliferation of hepatoma cells; (4) effects of inhibiting of GRP78 expression on the gene and protein expression of p53, p21, CDK2, CDK4, and CDK6 in hepatoma cells; (5) effects of HA15 on the proliferation and the gene and protein expression of p53, p21, CDK2, CDK4, and CDK6 in hepatoma cells. Measurement data of the normal distribution were expressed as Mean± SD, and comparison of groups was conducted using the t test or ANOVA. Repeated measurement data were analyzed using repeated ANOVA. Count data were expressed as absolute numbers, and comparisons between groups was conducted using the chi-square test. COX proportional hazards regression model was used for univariate and multivariate analysis. The Kaplan-Meier method was used to calculate the survival time and draw survival curve, and the Log-rank test was used for generative analysis. Results:(1) GRP78 expression in hepatocellular carcinoma tissue and adjacent tissue and its correlation with the clinicopathological characteristics of hepatocellular carcinoma patients: results of immunohistochemical staining of hepatocellular carcinoma tissue chip showed that GRP78 was low-expressed in 53 cases and high-expressed in 37 cases of the 90 hepatocellular carcinoma tissues. GRP78 was low-expressed in 84 cases and high-expressed in 6 cases of the 90 paracancerous tissues. There was a significant difference in GRP78 expression between hepatocellular carcinoma tissues and paracancerous tissues ( P<0.05). (2) Analysis of factors affecting the prognosis of hepatocellular carcinoma patients: all 90 patients were followed up for 5 to 56 months, with a median follow-up time of 49 months. The median overall survival time and median disease progression-free survival time were 56 months and 53 months in the 53 hepatocellular carcinoma patients with GRP78 as low-expressed, versus 32 months and 19 months in the 37 hepatocellular carcinoma patients with GRP78 as high-expressed, respec-tively, showing significant differences ( χ2=17.482, 12.097, P<0.05). Results of univariate analysis showed that alanine aminotransferase (ALT), tumor pathological grading and GRP78 expression were related factors affecting the 3-year overall survival rate and disease progression-free survival rate of hepatocellular carcinoma patients ( hazard ratio=2.317, 2.039, 3.740 and 2.194, 2.177, 2.927, 95% confidence interval as 1.150?4.671, 1.201?3.462, 2.116?6.612 and 1.048?4.593, 1.093?4.336, 1.492?5.742, P<0.05). Results of multivariate analysis showed that ALT >40 U/L, tumor pathological grading as Ⅲ-Ⅳ grade and GRP78 as high-expressed were independent risk factors affecting the 3-year overall survival rate and disease progression-free survival rate of hepatocellular carcinoma patients ( hazard ratio=2.438, 2.245, 3.223 and 3.046, 2.473, 3.307, 95% confidence interval as 1.114?5.334, 1.047?4.814, 1.396?7.440 and 1.337?6.940, 1.141?5.360, 1.399?7.819, P<0.05). (3) Effects of inhibiting of GRP78 expression on the proliferation of hepatoma cells: ①results of qRT-PCR showed that the relative expression of GRP78 messenger RNA (mRNA) in Huh7, Hep3B, and LO2 cells were 3.06±0.33, 4.42±0.60 and 1.00±0.02. There were significant differences in GRP78 mRNA expression between Huh7 and LO2 cells or Hep3B and LO2 cells ( t=6.19, 5.42, P<0.05). ②Results of Western Blot detection showed that the relative expression of GRP78 protein in Huh7, Hep3B, and LO2 cells were 1.65±0.01, 1.77±0.01 and 0.99±0.02. There were significant differences in GRP78 protein expression between Huh7 and LO2 cells or Hep3B and LO2 cells ( t=75.09, 108.10, P<0.05). ③Results of cell proliferation experiments showed that the growth rates in Hu7 GRP78 gene-specific shRNA lentiviruses group cells and Hu7 negative control shRNA lentivirus group cells at 24, 48, 72 and 96 hours were 111.51%±0.35%, 144.85%±0.68%, 188.71%±3.62%, 282.51%±5.25% and 190.08%±0.58%, 285.76%±2.69%, 459.51%±4.29%, 597.88%±12.25%, showing signifi-cant differences ( Fgroups=1 360.000, Ftime=668.500, Finteraction=197.600, P<0.05). The growth rates in Hep3B GRP78 gene-specific shRNA lentiviruses group cells and Hep3B negative control shRNA lentivirus group cells at 24, 48, 72 and 96 hours were 124.47%±0.25%, 153.25%±1.25%, 195.45%±3.19%, 282.51%±10.76% and 179.69%±0.33%, 322.67%±2.46%, 486.27%±5.82%, 622.35%±12.58%, showing significant differences ( Fgroups=1 222.000, Ftime=706.200, Finteraction=179.600, P<0.05). ④Results of the cell clone formation experiments showed that the number of cells in Hu7 GRP78 gene-specific shRNA lentiviruses group cells and Hu7 negative control shRNA lentivirus group cells were 125±3 and 435±17, showing a significant difference ( t=17.86, P<0.05). The number of cells in Hep3B GRP78 gene-specific shRNA lentiviruses group cells and Hep3B negative control shRNA lentivirus group cells were 138±3 and 388±7, showing a significant difference ( t=32.29, P<0.05). (4) Effects of inhibiting of GRP78 expression on the gene and protein expression of p53, p21, CDK2, CDK4, and CDK6 in hepatoma cells: results of high-throughput transcription histological analysis showed that the relative expression rates of p53, p21, CDK2, CDK4, and CDK6 were 19%, 334%, 398%, 41% and 49% in the Hu7 GRP78 gene-specific shRNA lentiviruses group cells comparing to the Hu7 negative control shRNA lentivirus group cells. ①Results of qRT-PCR showed that the relative expression of GRP78, p53, p21, CDK2, CDK4, and CDK6 mRNA were 0.17±0.03, 4.05±0.71, 3.73±0.47, 0.49±0.09, 0.48±0.06, 0.36±0.07 in the Hu7 GRP78 gene-specific shRNA lentiviruses group cells, versus 1.00±0.05, 1.03±0.17, 1.00±0.07, 1.01±0.09, 1.02±0.14, 1.00±0.03 in the Hu7 negative control shRNA lentivirus group cells, showing significant differences ( t=14.62, 4.17, 5.72, 4.26, 3.49, 8.82, P<0.05). The relative expression of GRP78, p53, p21, CDK2, CDK4, and CDK6 mRNA were 0.11±0.01, 4.28±0.43, 4.19±0.22, 0.44±0.01, 0.25±0.03, 0.68±0.04 in Hep3B GRP78 gene-specific shRNA lentiviruses group cells, versus 1.01±0.09, 1.02±0.15, 1.00±0.06, 1.01±0.09, 1.01±0.08, 1.15±0.02 in Hep3B negative control shRNA lentivirus group cells, showing significant differences ( t=10.19, 7.14, 13.79, 6.37, 9.42, 9.61, P<0.05). ②Results of Western Blot detection showed that the relative expression of GRP78, p53, p21, CDK2, CDK4, and CDK6 protein were 0.45±0.01, 1.98±0.05, 2.31±0.12, 0.75±0.03, 0.69±0.04, 0.82±0.03 in the Hu7 GRP78 gene-specific shRNA lentiviruses group cells, versus 1.01±0.05, 1.03±0.01, 1.00±0.02, 1.00±0.01, 1.01±0.02, 1.00±0.03 in the Hu7 negative control shRNA lentivirus group cells, showing significant differences ( t=11.07, 14.56, 11.30, 11.29, 10.55, 11.37, P<0.05). The relative expression of GRP78, p53, p21, CDK2, CDK4, and CDK6 protein were 0.61±0.03, 1.98±0.16, 2.55±0.12, 0.85±0.03, 0.78±0.01, 0.54±0.02 in Hep3B GRP78 gene-specific shRNA lentiviruses group cells, versus 1.00±0.03, 1.05±0.02, 1.05±0.01, 1.05±0.02, 1.00±0.02, 1.00±0.02 in Hep3B negative control shRNA lentivirus group cells, showing significant differences ( t=10.97, 13.40, 12.35, 11.06, 12.45, 13.78, P<0.05). (5) Effects of HA15 on the proliferation and the gene and protein expression of p53, p21, CDK2, CDK4, and CDK6 in hepatoma cells: results of 50% inhibiting concentration (IC50) test of HA15 showed that the IC50 of HA15 for Huh7 and Hep3B cells at 48 hours were 9.98 μmol/L and 13.70 μmol/L. ①Huh7 and Hep3B cells were treated with 9.98 μmol/L and 13.70 μmol/L of HA15. Results of cell proliferation experiments showed that the growth rates at 24, 48, 72, and 96 hours were 112.81%±0.27%, 154.71%±1.45%, 237.66%±16.77%, 294.40%±14.92% in the HA15-Huh7 cells, versus 133.67%±0.49%, 352.93%±2.31%, 557.17%±4.89%, 662.60%±13.31% in the normal Huh7 cells, showing a significant difference ( Fgroups=766.800, Ftime=518.200, Finteraction=133.300, P<0.05). The growth rates at 24, 48, 72, and 96 hours were 121.27%±2.32%, 203.85%±3.18%, 240.80%±3.02%, 286.50%±7.10% in the HA15-Hep3B cells, versus 239.14%±1.02%, 362.00%±5.44%, 539.37%±10.80%, 694.79%±17.13% in the normal Hep3B cells, showing a signifi-cant difference ( Fgroups=594.300, Ftime=317.900, Finteraction=78.600, P<0.05). ②Results of qRT-PCR showed that the relative expression of GRP78, p53, p21, CDK2, CDK4, and CDK6 mRNA were 0.27±0.05, 3.64±0.28, 4.13±0.41, 0.51±0.07, 0.39±0.03, 0.17±0.02 in the HA15-Huh7 cells, versus 1.02±0.14, 1.00±0.03, 1.00±0.05, 1.01±0.08, 1.01±0.09, 1.03±0.17 in the normal Huh7 cells, showing significant differences ( t=5.00, 9.25, 7.63, 4.73, 6.82, 5.01, P<0.05). The relative expression of GRP78, p53, p21, CDK2, CDK4, and CDK6 mRNA were 0.28±0.03, 3.49±0.78, 4.31±0.53, 0.38±0.05, 0.36±0.04, 0.24±0.03 in the HA15-Hep3B cells, versus 1.01±0.11, 1.03±0.18, 1.01±0.08, 1.00±0.06, 1.02±0.15, 1.00±0.06 in the normal Hep3B cells, showing significant differences ( t=6.26, 3.08, 6.21, 7.97, 4.26, 11.08, P<0.05). ③Results of Western Blot detection showed that the relative expression of GRP78, p53, p21, CDK2, CDK4, and CDK6 protein were 0.52±0.05, 1.94±0.08, 1.58±0.02, 0.89±0.00, 0.86±0.02, 0.74±0.01 in the HA15-Huh7 cells, versus 1.02±0.03, 1.00±0.03, 1.02±0.02, 1.04±0.03, 1.00±0.01, 1.01±0.02 in the normal Huh7 cells, showing significant differences ( t=11.54, 10.28, 11.03, 12.81, 13.67, 10.09, P<0.05). The relative expression of GRP78, p53, p21, CDK2, CDK4, and CDK6 protein were 0.57±0.02, 1.67±0.04, 1.41±0.04, 0.82±0.03, 0.70±0.02, 0.74±0.01 in the HA15-Hep3B cells, versus 1.03±0.01, 0.98±0.03, 1.00±0.03, 1.03±0.03, 1.01±0.01, 1.04±0.01 in the normal Huh7 cells, showing significant differences ( t=10.81, 11.54, 12.26, 13.62, 14.23, 10.17, P<0.05). Conclusions:High expression of GRP78 is an independent risk factor affecting the overall survival and disease progression-free survival of hepatocellular carcinoma patients. Inhibiting of GRP78 expression can reduce cell proliferation and the expression of p53, p21, CDK2, CDK4, and CDK6 mRNA and proteins in hepatoma cells.