ObjectiveTo investigate the effect of Homebox A6 （HOXA6） on the proliferation， invasion， metastasis， and apoptosis of HepG2 hepatoma cells and its association with the PI3K/AKT signaling pathway. MethodsHepG2 hepatoma cells were cultured， and HOXA6 overexpression plasmid and siRNA were constructed and transfected into cells. The cells were randomly divided into empty plasmid group， HOXA 6 overexpression group， siRNA negative control group， and siRNA HOXA6 interference group. CCK8 assay was used to measure cell proliferation， Transwell assay was used to observe cell invasion， and wound healing assay was used to observe cell migration （related proteins TIMP3， MMP9， and MMP3）. Flow cytometry was used to measure cell apoptosis （related proteins BAX and BCL2）， the BCA method was used to measure protein concentration， and Western Blot was used to measure the expression of related proteins. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the SNK-q test was used for further comparison between two groups. ResultsCompared with the empty plasmid group， HOXA6 overexpression significantly promoted the proliferation， invasion， and migration of HepG2 hepatoma cells （all P<0.001）， and there was a significant reduction in the protein expression of TIMP3 （P<0.001）， while there were significant increases in the expression levels of MMP9 and MMP3 （both P<0.001）. Compared with the siRNA negative control group， HOXA6 interference significantly inhibited the proliferation， invasion， and migration of HepG2 hepatoma cells （all P<0.001）， and there was a significant increase in the protein expression of TIMP3 （P<0.001）， while there were significant reductions in the expression levels of MMP9 and MMP3 （both P<0.001）. Flow cytometry showed that compared with the empty plasmid group， HOXA6 overexpression inhibited the apoptosis of HepG2 hepatoma cells （P<0.001）， with a significant reduction in the expression of the apoptosis-related protein BAX and a significant increase in the expression of BCL2 （both P<0.001）. Compared with siRNA negative control group， HOXA6 interference promoted the apoptosis of HepG2 hepatoma cells （P<0.001）， with a significant increase in the expression of BAX and a significant reduction in the expression of BCL2 （both P<0.001）. Compared with the empty plasmid group， the HOXA6 overexpression group had significantly higher ratios of p-AKT/AKT and p-PI3K/PI3K （both P<0.001）， and compared with the siRNA negative control group， the siRNA HOXA6 interference group had significantly lower ratios of p-AKT/AKT and p-PI3K/PI3K （both P<0.001）. ConclusionHOXA6 can promote the proliferation， invasion， and metastasis of HepG2 hepatoma cells and inhibit their apoptosis by activating the PI3K/AKT signaling pathway through phosphorylation.

Objective Based on a complete Freund's adjuvant(CFA)-induced chronic inflammatory pain model,we compared and analyzed the differences in anti-inflammatory and analgesic effects of moxibustion intervention initiated at different timepoints,aiming to identify the optimal timing for moxibustion intervention.The goal is to establish standardized intervention protocols for basic research on the anti-inflammatory and analgesic effects of moxibustion.Methods Male C57BL/6 mice were randomly divided into 3 groups based on the moxibustion initiation timepoints of 4,7,and 10 d after modeling.Then,the mice in each group were randomly assigned to 3 subgroups,including a control group,a model group,and a moxibustion group,with 8 mice in each subgroup.Chronic inflammatory pain was induced by injecting 20 μL of CFA into the sole of the right hind paw.Moxibustion applied at the"Zusanli"acupoint for 30 minutes started on the 4th,7th,and 10th days after modeling,and the intervention continued for 7 days.The latency of paw withdrawal to thermal radiation was measured to evaluate the pain threshold before modeling,after modeling,and on the 1st,4th,and 7th days of treatment.Foot volume was measured to assess toe swelling before modeling,after modeling,and on the 1st and 7th days of treatment.Results Compared with the control group,the model group exhibited a reduced pain threshold(P＜0.0001)and increased paw volume(P＜0.0001).Compared with the model group,the subgroups receiving moxibustion intervention initiated on the 4th,7th,and 10th days post-modeling exhibited an increased pain threshold(P＜0.05,P＜0.0001).However,the paw volume of the subgroups receiving moxibustion intervention initiated on the 4th day post-modeling increased(P＜0.0001),while those of the subgroups receiving moxibustion intervention initiated on the 7th and 10th days post-modeling decreased(P＜0.0001).Among the intervention subgroups receiving moxibustion initiated on days 4,7,and 10,the day 7 intervention-initiating subgroup showed significant increase in pain threshold(P＜0.05,P＜0.0001),and the day 7 and day 10 intervention-initiating subgroups showed significantly reduced paw volume(P＜0.0001).Conclusion Considering both the analgesic and anti-inflammatory effects of moxibustion,day 7 post-modeling may be the optimal time for moxibustion to achieve effective anti-inflammatory and analgesic outcomes.

Objective To analyze the level of 25-hydroxyvitamin D[25(OH)D]in patients with type 2 diabetes mellitus,and preliminarily investigate the correlation between serum 25(OH)D level and metabolic indicators such as glycosylated hemoglobin(HbA1c)and pancreatic islet function in patients with type 2 diabetes mellitus.Methods A total of 459 patients with type 2 diabetes mellitus who were hospitalized in the Department of Endocrinology,Xinxiang First People's Hospital from January 2020 to December 2020 were selected as the research subjects.Clinical data of the patients were collected,including gender,age,serum 25(OH)D,fasting insulin,C-peptide,HbA1c,fasting blood glucose,postprandial blood glucose,urinary microalbumin,urinary albumin-to-creatinine ratio,blood calcium,blood uric acid(UA),triglyceride(TG),total cholesterol(TCH),low-density lipoprotein(LDL),and high-density lipoprotein(HDL).According to the serum 25(OH)D level,the patients were divided into sufficient group[n=20,25(OH)D≥30 μg·L-1],insufficient group[n=95,20 μg·L-125(OH)D＜30 μg·L-1],deficient group[n=231,10 μg·L-1 ≤25(OH)D＜20 μg·L-1],and severely deficient group[n=113,25(OH)D＜10 μg·L-1].Differences in metabolic indicators of patients in the four groups were compared,and the correlation between 25(OH)D level and metabolic indicators was analyzed by using the Pearson correlation.Results The serum 25(OH)D level of patients with type 2 diabetes mellitus was 3.00-46.59(15.75±0.35)μg·L-1;the serum 25(OH)D level of male patients was significantly higher than that of female patients(P＜0.05).The prevalence of 25(OH)D deficiency in patients with type 2 diabetes mellitus was 74.9％(344/459).The 25(OH)D deficiency mainly occurred in January,February,March,April,November,and December.Patients in the insufficient,deficient,and severely deficient groups had significantly higher HbA1c levels than those in the sufficient group(P＜0.05),and the HbA1c levels of patients in the deficient and severely deficient groups were significantly higher than those in the insufficient group(P＜0.05);there was no statistically significant difference in the HbA1c level between the deficient group and the severely deficient group(P＞0.05).There was no statistically significant difference in fasting blood glucose between the sufficient group and insufficient group,and between the deficient group and severely deficient group(P＞0.05);fasting blood glucose of patients in the deficient and severely deficient groups was significantly higher than that in the sufficient and insufficient groups(P＜0.05).There was no statistically significant difference in fasting insulin,urinary microalbumin,daily total urinary albumin,and urinary albumin-to-creatinine ratio of patients among the sufficient,insufficient,and deficient groups(P＞0.05);the fasting insulin of patients in the severely deficient group was significantly lower than that in the sufficient,insufficient,and deficient groups(P＜0.05);the urinary microalbumin,daily total urinary albumin,and urinary albumin-to-creatinine ratio of patients in the severely deficient group were significantly higher than those in the sufficient,insufficient,and deficient groups(P＜0.05).There was no statistically significant difference in the homeostasis model assessment for insulin resistance(HOMA-IR),serum albumin,blood creatinine,1-hour postprandial blood glucose,2-hour postprandial blood glucose,3-hour postprandial blood glucose,fasting C-peptide,1-hour postprandial C-peptide,2-hour postprandial C-peptide,3-hour postprandial C-peptide,TG,TCH,LDL,HDL,blood UA,and blood calcium of patients among the four groups(P＞0.05).Pearson correlation analysis showed that serum 25(OH)D levels in patients with type 2 diabetes mellitus were negatively correlated with HbA1c,urinary microalbumin,and urinary albumin-to-creatinine ratio(r=-0.093,-0.166,-0.157;P＜0.05),and positively correlated with fasting insulin(r=0.089,P＜0.05).Serum 25(OH)D levels in patients with type 2 diabetes mellitus had no correlation with fasting blood glucose,HOMA-IR,serum albumin,blood creatinine,1-hour postprandial blood glucose,2-hour postprandial blood glucose,3-hour postprandial blood glucose,fasting C-peptide,1-hour postprandial C-peptide,2-hour postprandial C-peptide,3-hour postprandial C-peptide,TG,TCH,LDL,HDL,blood UA,and blood calcium(P＞0.05).Conclusion 25(OH)D deficiency and insufficiency are common in patients with type 2 diabetes mellitus,especially in female patients.In patients with type 2 diabetes mellitus,25(OH)D level is positively correlated with fasting insulin and negatively correlated with HbAlc,urinary microalbumin,and urinary albumin-to-creatinine ratio.25(OH)D deficiency in patients with type 2 diabetes mellitus is mainly distributed in January,February,March,April,November,and December.

OBJECTIVE To explore the influence of reusing and recycling times on the quality of ethanol recovered from ethanol deposition solution of aqueous extract of Lonicerae Japonicae Flos. METHODS The ethanol in five consecutive batches of Lonicerae Japonicae Flos water extract ethanol deposition was recovered by vacuum distillation. The impurities in five batches of recovered ethanol and the recovered ethanol when partial coking caused by temperature runaway were qualitatively analyzed by headspace solid phase micro extraction combined with gas chromatography-mass spectrometry(HS-SPME/GC-MS), and the ultraviolet absorbance of each batch of recovered ethanol was also analyzed. RESULTS Styrene was the only impurity in the first and second batch of recovered ethanol; there were five impurities in the third and fourth batch of recovered ethanol; the number of impurities in the fifth batch of recovered ethanol increases to 17. When partial coking was occurred, the exogenous impurities in the recovered ethanol increased dramatically to forty-one, and there were harmful substances such as ninhydrin and naphthalene. The ultraviolet absorbance of the third to fifth batch of recovered ethanol exceed the upper limit stipulated in the second part of Chinese Pharmacopoeia, 2020 Edition. While the ultraviolet absorbance of the recovered ethanol when partial coking takes place exceeds the standard of the pharmacopoeia greatly. CONCLUSION The recovered ethanol should be treated after being reused two times, so that its ultraviolet absorbance level and impurity type are equivalent to that of fresh ethanol before it can be used again. The method provides reference for the quality control and compliance use of recovered ethanol.

Intranasal drug administration can avoid the blood-brain barrier, gastrointestinal degradation and liver first-pass effect, and may directly deliver drugs to the brain through the olfactory nerve pathway, so it is being used for the treatment of various neurological diseases. This article reviewed the research progress of intranasal drug delivery in terms of nasal anatomy, transport pathways of intranasal drug delivery for central nervous system diseases, mechanisms of action, clinical application, challenges and novel technologies.

ObjectiveTo investigate the effect of Zhizi Dahuang decoction （ZZDHT） in the treatment of alcoholic liver disease （ALD） by improving oxidative stress in hepatic neutrophils. MethodsNetwork pharmacology was used to obtain the chemical components of ZZDHT and their corresponding action targets and analyze the potential targets and functional pathways of ZZDHT in the treatment of ALD. The non-target metabolomics technology was used to observe the changes in the metabolites of ZZDHT in mouse serum and liver. The mice were given ZZDHT at a dose twice as much as the middle dose concentration by gavage， and serum and liver samples were collected at six time points after gavage （10 minutes， 30 minutes， 1 hour， 2 hours， 4 hours， and 6 hours） and were then mixed for mass spectrometry （administration group with 18 mice）， while the 18 mice in the control group were given an equal volume of normal saline by gavage. Ultra-performance liquid chromatography was used for rapid isolation and identification of the metabolites of ZZDHT in serum and liver tissue， and the effective constituents of ZZDHT were validated. Male C57BL/6J mice， aged 8 weeks， were randomly and equally divided into control group， model group， and low-， middle-， and high-dose ZZDHT groups， with 10 mice in each group. All mice except those in the control group were used to establish a mouse model of ALD （NIAAA model mice）， and at the same time， the mice in the administration groups were given low-， middle-， and high-dose ZZDHT by gavage， while those in the control group and the model group were given an equal volume of normal saline by gavage. The serum levels of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， and triglyceride （TG） were measured； PCR was used to measure the gene expression levels of related inflammation， oxidative stress， and neutrophil indicators in the liver； ELISA was used to measure the levels of related inflammation and oxidative stress indicators in serum； superoxide dismutase （SOD）， glutathione peroxidase （GSH-Px）， and malondialdehyde （MDA） were measured to observe the level of oxidative stress in the liver； HE staining， myeloperoxidase staining， and oil red staining were used to observe liver injury， neutrophil infiltration， and lipid deposition. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsA total of 53 active components and 227 target genes were obtained for ZZDHT， and there were 8685 target genes of ALD， resulting in 222 common target genes between these two groups of genes. Core pathways included the interleukin-6 signaling pathway and the TNF signaling pathway. The non-targeted metabolic analysis of ZZDHT obtained 225 metabolites in mouse liver and 227 metabolites in serum， among which there were 126 common metabolites. The core pathways of liver metabolites included glycerolipid metabolism and inflammatory mediator regulation of TRP channels， and the core pathways of serum metabolites included the AMPK signaling pathway and oxidative phosphorylation， all of which were associated with oxidative stress- and inflammation-related pathways. Compared with the model group， the low-， middle-， and high-dose ZZDHT groups had significant reductions in the serum levels of ALT， AST， and TG （all P<0.05）， and the middle-dose ZZDHT group had significant reductions in the levels of Ly6g， Ncf1， Ncf2， IL-6， TNF-α， IL-1β， MDA， 4-HNE， Gp91， and P22 in the liver （all P<0.05）， a significant increase in the level of SOD （P<0.05）， a significant reduction in the serum level of 4-HNE （P<0.05）， and a significant increase in the level of GSH-Px （P<0.05）. There were significant improvements in fat deposition and neutrophil infiltration in the liver of mice in the middle-dose ZZDHT group （both P<0.05）. ConclusionZZDHT significantly reduces oxidative stress and inflammatory response in NIAAA model mice.

Nasal preparations have unique advantages in drug delivery and are widely used in the treatment of local and systemic diseases. Nasal administration of traditional Chinese medicine （TCM） has a long history in China. In recent years， nasal preparations of TCM have attracted wide attention. Based on the information about nasal preparations of TCM from the database of National Medical Products Administration （NMPA）， Yaozh.com and China National Knowledge Internet （CNKI） in the recent 30 years， the formulation， the listed products， commonly used TCM， pharmaceutical excipients， clinical application and safety research of modern nasal preparations of TCM were summarized and expounded. Focusing on many problems in the development of modern nasal preparations of TCM， such as inaccurate dosage of some products， incomplete quality standard system of pharmaceutical excipients， imperfect safety evaluation， lack of research and development of nasal drug delivery devices and so on， the possible solutions and prospects were put forward from the aspects of optimizing the extraction and separation process of TCM， the quality control and application method of pharmaceutical excipients， the development of new dosage forms， the safety evaluation of nasal administration of TCM， and the design and development of nasal administration devices. The aim is to provide ideas for the development of nasal preparations of TCM and provide scientific basis for its sustainable utilization.

Objective To investigate the effect of atractylone on the viability and apoptosis of hepatoma HepG2 cells and its mechanism of action. Methods Hepatoma HepG2 cells were selected and divided into low-, middle-, and high-dose atractylone groups (5, 10, and 20 μmol/L), and the cells in the control group were added with an equal volume of DMSO. MTT colorimetry was used to measure the viability of HepG2 cells after treatment with different concentrations of atractylone; flow cytometry was used to measure the apoptosis rate and mitochondrial membrane potential of HepG2 cells; the DCFH-DA fluorescent probe labeling method was used to measure the level of reactive oxygen species (ROS) in HepG2 cells; Transwell assay was used to evaluate the effect of atractylone on the migration ability of HepG2 cells; Western blot was used to measure the protein expression levels of Bcl-2, Bax, and cleaved caspase-3. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t -test was used for comparison between two groups. Results After 24 and 48 hours of treatment with atractylone, compared with the control group, the low-, middle-, and high-dose atractylone groups had a tendency of reduction in cell viability (all P < 0.05), with a half inhibitory concentration of 26.19 μmol/L in atractylone treatment of HepG2 cells for 72 hours. The low-, middle-, and high-dose atractylone groups had a significantly higher apoptosis rate than the control group (14.34%/29.32%/50.12% vs 0.32%, all P < 0.05). Compared with the control group, the low-, middle-, and high-dose atractylone groups had a significant increase in the fluorescence intensity of ROS in HepG2 cells (all P < 0.05). After 48 hours of treatment with atractylone, compared with the control group, the low-, middle-, and high-dose atractylone groups had a significant reduction in the number of migrated cells (132.67±18.36/57.00±9.26/31.00±2.45 vs 258.11±38.54, P < 0.05). Compared with the control group, the low-, middle-, and high-dose atractylone groups had a significant reduction in the expression of the anti-apoptotic factor Bcl-2 and significant increases in the expression of the apoptotic factors Bax and cleaved caspase-3 (all P < 0.05). Conclusion Atractylone can induce the apoptosis and inhibit the migration of HepG2 cells, which provides an experimental basis for further development and utilization of atractylone.

Objective:To compare the efficacy and safety of Tonghua Dongbao′s insulin aspart injection (Rishulin) and NovoRapid (Novo Nordisk) in the treatment of diabetes.Methods:A 26-week, randomized, open-label, parallel-group, positive control drug and non-inferiority trial was conducted in 23 centers in China. A total of 563 diabetes with poor blood glucose control treated with insulin for at least 3 months before were included. The subjects were randomized(stratified block random method) into those receiving Rishulin or NovoRapid at a ratio of 3∶1. Both groups were combined with basal insulin (Lantus). The primary endpoint was the change in glycosylated hemoglobin (HbA1c) from baseline to the end of 24 weeks of treatment.Results:For full analysis set, after 24 weeks of treatment, HbA1c level of Ruishulin group decreased from (8.66±1.28)% to (7.77±1.09)% ( P<0.001), and that of NovoRapid group decreased from (8.47±1.28) % to (7.65±0.97) % ( P<0.001). Treatment difference in HbA1c (NovoRapid group-Ruishulin group) was -0.061% (95% CI -0.320-0.199). HbA1c<7.0% target reacing rates were 24.26% and 21.21% ( P=0.456), and HbA1c<6.5% target reacing rates were 9.65% and 6.82% ( P=0.310) in Ruishulin group and NovoRapid group, repectively. The standard 2 hours postprandial blood glucose (2hPG) in Ruishulin group decreased from (16.23±5.22) mmol/L to (12.65±4.57) mmol/L ( P<0.001), and 2hPG in NovoRapid group decreased from (16.13±5.37) mmol/L to (11.91)±4.21) mmol/L ( P<0.001). The fingertips blood glucose at 7-point of both groups exhibited varying degrees of reduction compared with those at baseline, repectively. Positive ratios of specific antibodies were 31.68% in Ruishulin group and 36.36% in NovoRapid group ( P=0.320). Ratios of negative to positive were 7.43% and 10.61% ( P=0.360), and ratios of positive to negative were 10.40% and 7.58% ( P=0.360) in Ruishulin group and NovoRapid group, respectively. The incidence of hypoglycemia was 60.05% and 55.40% ( P=0.371), and the incidence of adverse events was 76.60% and 77.70% ( P=0.818) in Ruishulin group and NovoRapid group, respectively. Conclusions:Rishulin is not inferior to NovoRapid, and has shown good efficacy and safety. It can be an ideal choice for clinicians in patients with poor blood glucose control with insulin.

OBJECTIVE: To explore the action mechanisms of Huangqi Decoction Granules (, HQDG) on hepatitis B cirrhosis. METHODS: A total of 85 patients with hepatitis B cirrhosis were randomly divided into HQDG group (42 cases) and control group (43 cases) by a random number table and were treated with HQDG or placebo for 48 weeks (6 g per times and orally for 3 times a day), respectively. After RNA-sequencing of serum samples extracted from the patients, the differentially expressed genes (DEGs) in HQDG and control groups before and after treatment were separately screened. The DEGs were then performed pathway enrichment analysis and proteinprotein interaction (PPI) network analysis. The expression levels of key genes were detected by quantitative realtime polymerase chain reaction (qRT-PCR). RESULTS: After the investigation, 4 and 3 cases were respectively excluded from HQD and control groups because of the incomplete data. Additionally, 3 and 5 cases were lost to follow up in HQD and control groups respectively. Finally, a total of 70 cases with good compliance were included for further DEGs analysis. A total of 1,070 DEGs (including 455 up-regulated genes and 615 down-regulated genes) in HQDG group and 227 DEGs (including 164 up-regulated genes and 63 down-regulated genes) in the control group were identified after treatment. Compared with the control group, 1,043 DEGs were specific in HQDG group. Besides, 1 up-regulated transcription factor (TF, such as GLI family zinc finger 1, GLI1) and 25 down-regulated TFs (such as drosophila mothers against decapentaplegic proteinfamily member 2, SMAD2) were identified. Pathway enrichment analysis showed that down-regulated Ras homolog gene family member A (RHOA) was enriched in pathogenic Escherichia coli infection. In the PPI network, up-regulated epidermal growth factor receptor (EGFR), and down-regulated cell division cycle 42 (CDC42) as well as v-akt murine thymoma viral oncogene homolog 1 (AKT1) had higher degrees. Moreover, long non-coding RNAs (lncRNA) growth arrest-specific 5 (GAS5) was involved in the lncRNA-target regulatory network. Furthermore, qRT-PCR revealed that expression levels of CDC42 and GLI1 had significant differences in HQDG group before and after treatment (P<0.05). CONCLUSIONS CDC42 and GLI1 may be the targets of HQDG in patients with hepatitis B cirrhosis. Additionally, SMAD2, EGFR, AKT1, RHOA and GAS5 might be associated with the curative effect of HQDG on hepatitis B cirrhosis patients.