TNF-α derived from M2 tumor-associated macrophages promotes epithelial-mesenchymal transition and cancer stemness through the Wnt/ß-catenin pathway in SMMC-7721 hepatocellular carcinoma cells.

M2-polarized tumor-associated macrophages (M2-TAMs) infiltrating the tumor microenvironment contribute to hepatocellular carcinoma (HCC) progression. It was reported that cancer cells undergoing EMT will acquire stemness characteristics. Here, the HCC SMMC-7721 cell line was co-cultured with M2-TAMs polarized from THP-1 cells in vitro. In in vivo studies, we used nude mice subcutaneous tumor model to test whether the growth of the tumor was affected by M2-TAMs. Subsequently, EMT, stemness and Wnt/ß-catenin pathway related markers were detected in cells and subcutaneous tumor tissues. TNF-α was also assessed in both the co-culture system supernatants and in nude mice serum. We found that SMMC-7721 underwent EMT and acquired stemness after co-culture with M2-TAMs, and resulted in larger tumor size following subcutaneous injection of SMMC-7721 suspended in M2-TAMs supernatants compared with SMMC-7721 alone. Enzyme linked immunosorbent assay showed that TNF-α expression was elevated in supernatants of M2-TAMs and positively correlated with tumor size in the serum of nude mice. Furthermore, we found that the Wnt/ß-catenin pathway was a downstream target of TNF-α and that the Wnt/ß-catenin inhibitor ICG-001 partially reversed EMT and attenuated cancer stemness. Our results indicate that TNF-α derived from M2-TAMs promote EMT and cancer stemness cells via the Wnt/ß-catenin pathway.

Gut microbiota dynamics in KK-Ay mice: restoration following antibiotic treatment.

The primary aim of this study was to investigate the alterations in the microbial community of KK-Ay mice following antibiotic treatment. A comparative analysis of the gut microbiota was conducted between KK-Ay mice treated with antibiotics and those without treatment. The microbial community dynamics in antibiotic-treated KK-Ay mice were meticulously assessed over an eight-week period using 16S rDNA sequencing analysis. Simultaneously, dynamic renal function measurements were performed. The results demonstrated a marked decrease in bacterial DNA abundance following antibiotic intervention, coupled with a substantial reduction in bacterial diversity and a profound alteration in microbial composition. These observed microbiota changes persisted in the KK-Ay mice throughout the eight-week post-antibiotic treatment period. Particularly noteworthy was the reemergence of bacterial populations after two weeks or more, resulting in a microbiota composition resembling that of untreated KK-Ay mice. This transition was characterized by a significant increase in the abundance of clostridia at the class level, Lachnospirales and Oscillospirales at the order level, and Lachnospiraceae, Oscillospiraceae, and Ruminococcaceae at the family level. Concurrently, there was a notable decrease in Clostridia_UCG-014. The observed alterations in the gut microbiota of antibiotic-treated KK-Ay mice suggest a dynamic response to antibiotic intervention and subsequent restoration towards the original untreated state.

Specific Alternation of Gut Microbiota and the Role of Ruminococcus gnavus in the Development of Diabetic Nephropathy.

In this study, we aim to investigate the precise alterations in the gut microbiota during the onset and advancement of diabetic nephropathy (DN) and examine the impact of Ruminococcus gnavus (R. gnavus) on DN. Eight-week-old male KK-Ay mice were administered antibiotic cocktails for a duration of two weeks, followed by oral administration of R. gnavus for an additional eight weeks. Our study revealed significant changes in the gut microbiota during both the initiation and progression of DN. Specifically, we observed a notable increase in the abundance of Clostridia at the class level, higher levels of Lachnospirales and Oscillospirales at the order level, and a marked decrease in Clostridia_UCG-014 in DN group. Additionally, there was a significant increase in the abundance of Lachnospiraceae, Oscillospiraceae, and Ruminococcaceae at the family level. Moreover, oral administration of R. gnavus effectively aggravated kidney pathology in DN mice, accompanied by elevated levels of urea nitrogen (UN), creatinine (Cr), and urine protein. Furthermore, R. gnavus administration resulted in down-regulation of tight junction proteins such as Claudin-1, Occludin, and ZO-1, as well as increased levels of uremic toxins in urine and serum samples. Additionally, our study demonstrated that orally administered R. gnavus up-regulated the expression of inflammatory factors, including nucleotide-binding oligomerization domain-like receptor pyrin domain-containing protein 3 (NLRP3) and Interleukin (IL)-6. These changes indicated the involvement of the gut-kidney axis in DN, and R. gnavus may worsen diabetic nephropathy by affecting uremic toxin levels and promoting inflammation in DN.

An Update on the Role and Potential Molecules in Relation to Ruminococcus gnavus in Inflammatory Bowel Disease, Obesity and Diabetes Mellitus.

Ruminococcus gnavus (R. gnavus) is a gram-positive anaerobe commonly resides in the human gut microbiota. The advent of metagenomics has linked R. gnavus with various diseases, including inflammatory bowel disease (IBD), obesity, and diabetes mellitus (DM), which has become a growing area of investigation. The initial focus of research primarily centered on assessing the abundance of R. gnavus and its potential association with disease presentation, taking into account variations in sample size, sequencing and analysis methods. However, recent investigations have shifted towards elucidating the underlying mechanistic pathways through which R. gnavus may contribute to disease manifestation. In this comprehensive review, we aim to provide an updated synthesis of the current literature on R. gnavus in the context of IBD, obesity, and DM. We critically analyze relevant studies and summarize the potential molecular mediators implicated in the association between R. gnavus and these diseases. Across numerous studies, various molecules such as methylation-controlled J (MCJ), glucopolysaccharides, ursodeoxycholic acid (UDCA), interleukin(IL)-10, IL-17, and capric acid have been proposed as potential contributors to the link between R. gnavus and IBD. Similarly, in the realm of obesity, molecules such as hydrogen peroxide, butyrate, and UDCA have been suggested as potential mediators, while glycine ursodeoxycholic acid (GUDCA) has been implicated in the connection between R. gnavus and DM. Furthermore, it is imperative to emphasize the necessity for additional studies to evaluate the potential efficacy of targeting pathways associated with R. gnavus as a viable strategy for managing these diseases. These findings have significantly expanded our understanding of the functional role of R. gnavus in the context of IBD, obesity, and DM. This review aims to offer updated insights into the role and potential mechanisms of R. gnavus, as well as potential strategies for the treatment of these diseases.

Two weeks of high glucose intake is enough to induce intestinal mucosal damage and disturb the balance of the gut microbiota of rats.

High glucose plays a critical role in diabetes. However, the point when high glucose induces diabetes and the organ that triggers the initiation of diabetes remain to be elucidated. The aim of the present study was to clarify the damage induced on different organs of rats, when administered a 2-week infusion of dietary glucose. SD rats (12 weeks old) were randomly divided into normal diet, high glucose infusion (IHG) and oral high glucose (OHG) groups. The levels of fasting blood sugar, tumor necrosis factor (TNF)-α and interleukin (IL)-6 were assessed. Intestine, kidney and liver samples were collected for pathological examination. Feces were collected from the rats for gut microbiota assessment. The results indicated that short-term high glucose induced hyperglycemia that lasted for at least 2 weeks after cessation of high glucose intake. Short-term high glucose also clearly increased the serum levels of IL-6 and TNF-α, led to jejunum mucosa injury and obvious steatosis in hepatocytes, and disturbed the balance of the gut microbiota. OHG led to swelling and necrosis of individual intestinal villi. IHG led to the necrosis and disappearance of cells in the upper layer of the intestinal mucosa. The lesions were confined to the mucosa. A degree of glomerular cell swelling and apoptosis were also observed. Short-term high glucose intake induced lesions in the liver, kidney and intestine, disturbed the balance of the gut microbiota and may consequently induce diabetes complications.

Jiangtang Decoction Ameliorates Diabetic Kidney Disease Through the Modulation of the Gut Microbiota.

Purpose: This study aimed to elucidate the impact of Jiangtang decoction (JTD) on diabetic kidney disease (DKD) and its association with alterations in the gut microbiota. Methods: Using a diabetic mouse model (KK-Ay mice), daily administration of JTD for eight weeks was undertaken. Weekly measurements of body weight and blood glucose were performed, while kidney function, uremic toxins, inflammation factors, and fecal microbiota composition were assessed upon sacrifice. Ultra-structural analysis of kidney tissue was conducted to observe the pathological changes. Results: The study findings demonstrated that JTD improve metabolism, kidney function, uremic toxins and inflammation, while also exerting a modulatory effect on the gut microbiota. Specifically, the genera Rikenella, Lachnoclostridium, and unclassified_c_Bacilli exhibited significantly increased abundance following JTD treatment, accompanied by reduced abundance of norank_f_Lachnospiraceae compared to the model group. Importantly, Rikenella and unclassified_c_Bacilli demonstrated negative correlations with urine protein levels. Lachnoclostridium and norank_f_Lachnospiraceae were positively associated with creatinine (Cr), indoxyl sulfate (IS) and interleukin (IL)-6. Moreover, norank_f_Lachnospiraceae exhibited positive associations with various indicators of DKD severity, including weight, blood glucose, urea nitrogen (UN), kidney injury molecule-1 (KIM-1) levels, trimethylamine-N-oxide (TMAO), p-cresyl sulfate (pCS), nucleotide-binding oligomerization domain (Nod)-like receptor family pyrin domain-containing 3 (NLRP3) and IL-17A production. Conclusion: These findings suggested that JTD possess the ability to modulate the abundance of Rikenella, Lachnoclostridium, unclassified_c_Bacilli and norank_f_Lachnospiraceae within the gut microbiota. This modulation, in turn, influenced metabolic processes, kidney function, uremic toxin accumulation, and inflammation, ultimately contributing to the amelioration of DKD.

Specific alterations of gut microbiota in diabetic microvascular complications: A systematic review and meta-analysis.

Background: The role of gut microbiota in diabetes mellitus (DM) and its complications has been widely accepted. However, the alternation of gut microbiota in diabetic microvascular complications (DC) remains to be determined. Methods: Publications (till August 20th, 2022) on gut microbiota in patients with DC were retrieved from PubMed, Web of Science, Embase and Cochrane. Review Manager 5.3 was performed to estimate the standardized mean difference (SMD) and 95% confidence interval (CI) and calculate alpha diversity indices and the relative abundance of gut microbiota between patients in DC v.s. DM and DC v.s. healthy controls (HC). Results: We included 13 studies assessing 329 patients with DC, 232 DM patients without DC, and 241 HC. Compared to DM, patients with DC shared a significantly lower Simpson index (SMD = -0.59, 95% CI [-0.82, -0.36], p < 0.00001), but a higher ACE index (SMD = 0.42, 95% CI[0.11, 0.74], p = 0.009). Compared to HC, DC patients held a lower ACE index (SMD = -0.61, 95% CI[-1.20, -0.02], p = 0.04). The relative abundances of phylum Proteobacteria (SMD = 0.03, 95% CI[0.01, 0.04], p = 0.003, v.s. HC) and genus Klebsiella (SMD = 0.00, 95% CI[0.00, 0.00], p < 0.00001, v.s. HC) were enriched, accompanying with depleted abundances of phylum Firmicutes (SMD = -0.06, 95% CI[-0.11, -0.01], p = 0.02, v.s. HC), genera Bifidobacterium (SMD = -0.01, 95% CI[-0.02,-0.01], p < 0.0001, v.s. DM), Faecalibacterium (SMD = -0.01, 95% CI[-0.02, -0.00], p = 0.009, v.s. DM; SMD = -0.02, 95% CI[-0.02, -0.01], p < 0.00001, v.s. HC) and Lactobacillus (SMD = 0.00, 95% CI[-0.00, -0.00], p < 0.00001, v.s. HC) in DC. Conclusions: Gut microbiota perturbations with the depletion of alpha diversity and certain short-chain fatty acids (SCFAs)-producing bacteria were associated with the pathology of DC. Therefore, gut microbiota might serve as a promising approach for the diagnosis and treatment of DC. Further investigations are required to study the mechanisms by which gut dysbiosis acts on the onset and progression of DC.

[Effects of danhong injection on experimental atherosclerosis rabbit model and its mechanism].

OBJECTIVE: To observe the effects of Danhong injection on experimental atherosclerosis rabbits and explore its possible mechanism. METHODS: Forty New-Zealand male rabbits were randomly divided into four groups: normal control group, high-cholesterol group, danhong injection group and aspirin group, all rabbits were administered for fourteen weeks. The plaque area of aorta in each group was determined by image analysis. The blood lipid level was tested by ELISA method, malonaldehyde (MDA) in aorta wall was analyzed with thiobarbituric acid method, the iNOS and COX-2 in aorta wall was tested with RT-PCR method. RESULT: There were significant difference between danhong injection group and other groups (P < 0.5), danhong injection and Aspirin decreased the TG, TC and LDL-C level in plasma of rabbit model and decreased the expression of inflammatory cytokines and the oxidative stress level in aorta wall. CONCLUSION: Danhong injection can inhibit the formation of experimental atherosclerosis in rabbits, which may be related to decreasing blood lipid, plaque inflammation and oxidative stress level.

[Effect of paeonol on protecting endothelial cells of diabetic rats].

OBJECTIVE: To study the effect of paeonol on protecting endothelial cells of diabetic rats. METHODS: Streptozocin was used on rats to make diabetic models. Different dosages of paeonol were fed on all the model rats. PGI, TXA2, ET, CRP, ICAM-1, VCAM-1 and NO were tested after 30 day's therapy. RESULTS: Compared with control group PGI increased from 89.75 +/- 2.75, 89.97 +/- 7.28, 89.97 +/- 11.36 to 120.03 +/- 13.85, 108.34 +/- 11.25, 105.32 +/- 8.85, respectively. TXA2 decreased from 157.64 +/- 10.36, 156.64 +/- 11.35, 153.33 +/- 19.40 to 124.46 +/- 18.67, 136.40 +/- 18.15, 138.40 +/- 22.20, respectively. ET decreased from 181.68 +/- 5.10, 181.27 +/- 4.76, 181.04 +/- 4.19 to 140.55 +/- 3.01, 150.51 +/- 2.22, 161.02 +/- 3.76, respectively. CRP decreased from 41.63 +/- 7.37, 44.83 +/- 7.80, 42.06 +/- 7.21 to 28.62 +/- 5.45, 30.00 +/- 10.73, 37.09 +/- 4.94, respectively. ICAM-1 decreased from 225.77 +/- 11.96, 222.78 +/- 14.46, 225.17 +/- 10.03 to 190.93 +/- 12.67, 197.42 +/- 14.56, 200.64 +/- 15.36, respectively. VCAM-1 decreased from 566.72 +/- 24.46, 560.61 +/- 25.67, 359.61 +/- 42.75 to 506.26 +/- 37.26, 516.83 +/- 28.17, 527.02 +/- 43.47, respectively. NO had no change. CONCLUSIONS: Paeonol can protect the endothelial cells of diabetic rats.

[The summary of the clinic study of Zhi-Xin-Fang treating the heart failure resulting from cardiomyopathy].

OBJECTIVE: To observe the therapeutic and prognostic effects and to evaluate the safety of Zhi-Xin-Fang on heart failure resulting from cardiomyopathy. METHODS: 62 patients with cardiomyopathy combined with heart failure were treated with standard western medical therapy as treatment group. The other 60 patients with the same diseases were administered with both Zhi-Xin-Fang and standard western medicines as control. Several parameters were observed to evaluate the effect of Zhi-Xin-Fang on the heart failure, containing the therapeutic effect of clinical symptom and physical signs, the improvement of heart function and the ultrasonic caridogram. We also observed the effect of Zhi-Xin-Fang to relieve the clinical symptom and improve the heart function and quality of life and prognosis. RESULTS: As the NYHA classify for heart failure, the effective rate for the treatment group was 95% , more effective than the control group (effective rate: 80.65%) (P<0.05). There was significant improvement for LVEF and LVDS for both groups. However, the treatment group was better than the control group (P<0.05). The clinical symptom was significantly improved compared to the untreatment in both groups. The treatment group was better than the control group for effective rate (P<0.05) but not for the total effective rate. There was no significant difference for the function of liver and kidney, blood routine method, electrolyte compared to before for the two groups. CONCLUSION: The standard western medicines combined with Zhi-Xin-Fang is a good therapy for treating heart failure resulting from cardiomyopathy. The combined therapy can relieve the cinical symptom of heart failure, improve the heart function and quality of life. It is also safer for the patients. However, its long-term prognosis should be further studied in the future.

[Effect of cortex moutan on PGI2, TXA2, ET and NO in diabetic rats].

OBJECTIVE: To study the effect of Paeonol on PGI2, TXA2, ET and NO in diabetic rats. METHODS: Streptozocin was used on rats to make diabetic animal models. Different dosage of Paeonol were used on diabetic animal models. PGI2, TXA2, ET and NO were tested after 30 days therapy. RESULTS: Compared with those in the control group, the level of PGI2 increased and the contents of TXA2 and ET decreased in treatment group. Among them, the high dosage group was more obvious (P < 0.01). But the level of No Had no change. CONCLUSION: Paeonol can decrease the ET and TXA2 in diabetic rats, and increase PGI2 in diabetic rats.

[Effects of weikangning contained serum on growth of gastric cancer cell, expression of vascular endothelial growth factor and its receptors including KDR and fit-1].

OBJECTIVE: To observe the effect of drug-containing serum of Chinese traditional herbal decoction Weikangning (WKN) on growth of gastric cancer cell, expression of vascular endothelial growth factor (VEGF) and its receptors including KDR and Flt-1. METHODS: A total of 120 male Wistar rats were given high, medium and low-dose WKN. After the drug-containing serum was prepared, the gastric cancer cells MGC-803 of different dose groups were cultured with the drug-containing serum, respectively. The gastric cell growth was observed by using light microscope and flow cytometer,the expression of VEGF and its receptor Flt-1 was detected with SABC immunohistochemistry method and the mRNA expression levels of VEGF and its receptors including KDR and Flt-1 of different groups were detected with reverse transcription-polymerase chain reaction (RT-PCR), respectively. RESULTS: The gastric cancer cell growth and cell cycle of the three medicated groups were significantly improved as compared with those of the control group (P <0. 01) ,the proportion of cells in G0 - G1 phase was increased,while the cells in S phase were decreased. It was shown that the apoptotic rates were increased in the medicated groups in a dose-dependent manner. The gray scales ( microm(2) ) of VEGF and Flt-1 in high, medium and low dose groups was 182. 44 +/-0. 54,178. 65 +/-0. 56,174. 80 +/-0. 81 and 168. 51 +/- 0. 81,162. 01 +/-0. 52,148. 20 +/-0. 69, respectively vs 147.82 +/-0. 15(P <0.01) and 144.31 +/-0.71 (P <0.01) in the control group. The mRNA expression of VEGF and its receptors significantly decreased in gastric cancer cells after cultured with WKN contained serum (P < 0. 01 ). CONCLUSION: WKN has inhibitory effects on gastric cancer cell growth and mRNA expression of VEGF as well as its receptors KDR and Flt-1.

Effect of Weikangning on gastric cancer cell growth and expression of vascular endothelial growth factor and its receptors KDR and Flt-1.

AIM: To observe the effect of Chinese traditional herbal decoction Weikang-ning (WKN) on cell growth and expression of VEGF and its receptors KDR and Flt-1 in gastric cancer cell line MGC-803. METHODS: A total of 120 male Wistar rats were divided into control group, high dose, medium dose and low dose groups fed with natural saline, 20, 10, and 5 g/kg of WKN, respectively. The experimental animals were finally killed for the preparation of drug-containing serum. The gastric cancer cell MGC-803 was cultured with the drug-containing serum drawn from the rats in different groups. We observed the growth condition of the cancer cells with light microscope and flow cytometer. The expression of mRNA of VEGF and its receptors KDR and Flt-1 was detected with RT-PCR. RESULTS: The proportion of cells in G(0)-G(1) phase was (65.40+/-0.41)%, (56.92+/-0.62)%, (55.89+/-0.69)% in high, medium and low dose groups respectively vs (41.35+/-0.55)% in control group (P<0.01), while the cells in G(2)-S and S phases were (11.62+/-0.62)% and (22.99+/-0.69)%, (17.08+/-0.80)% and (26.00+/-0.71)%, (19.37+/-0.57)% and (24.74+/-0.64)% in high, medium and low dose groups, respectively, vs (23.65+/-0.56)% and (35.00+/-0.60)% in control group (P<0.01). The expression of mRNA of VEGF and its receptors was significantly decreased, the area of electrophoresis bands (AREA), the absorptivity of mean optical density (A) and the product of AREA and A were significantly lower in WKN-administered groups than that in control group (P<0.01). CONCLUSION: The decoction of WKN suppresses the growth of gastric cancer cell MGC-803 and decreases the expression of mRNA of both VEGF and its receptors KDR and Flt-1.

Irbesartan can improve blood lipid and the kidney function of diabetic nephropathy.

AIM: Diabetic nephropathy (DN) is the most frequent cause of end-stage renal disease. The activation of the renin-angiotensin system (RAS) and lipid disorders are major risk factors in progressive chronic kidney disease. Inhibition of the RAS is one of the most widely used therapies to treat chronic kidney disease. But its effect is not sufficient, and lowered hyperlipidemia is required. Most of medications for hypertension have effects only on the blood pressure in DN. This study is to evaluate the influence of irbesartan on blood lipid, kidney function, and the pathological change of kidney, liver, and adipose tissue. METHODS: Six-week old db/db mice were randomly assigned to control group and irbesartan group. Mice in irbesartan group were fed 40 mg/kg irbesartan each day. Eight weeks later, blood lipid, kidney function, and the pathological change of kidney, liver, and adipose tissue were measured. RESULTS: The results indicated that the blood lipid, uric acid, urea nitrogen, and creatinine of db/db mice increased significantly. There are obvious vacuolar degeneration and ballooned hepatocytes around the central vein of db/db mouse liver. Kidney biopsy found glomerular hypertrophy of glomerular, mesangial thickening, and vacuolar degeneration. Irbesartan can decrease the blood pressure, blood lipid, and kidney lipid. But it has no effects on blood glucose and liver lipid. It can improve the function and pathological change of kidney of db/db mice. But it has no effects on pathological change of adipose tissue and liver. CONCLUSIONS: Irbesartan can decrease blood lipid and protect the kidney of db/db mice, and is a good choice of treatment for diabetic nephropathy.

Moxibustion with Chinese herbal has good effect on allergic rhinitis.

Allergic rhinitis (AR) is a chronic inflammatory disease of rhino-ocular mucosa, affecting up to 40% of population worldwide. Chinese herbal medicines and Acupuncture, adopted thousands of years in China, has good effect on allergic rhinitis. This study evaluates the effects of Moxibustion with Chinese herbal in treating patients with allergic rhinitis over a 1-year follow-up. A randomized controlled trial was conducted in a sample of 355 participants recruited from Guangdong general hospital of China. After baseline measurements, participants were randomly assigned to treatment-group or control group. Treatment group received Moxibustion with Chinese herbal. Control group received Loratadine. The main outcomes, including symptom severity and quality of life were measured using the Allergic Rhinoconjunctivitis Quality of Life Questionnaire (RQLQ). Both moxibustion with Chinese herbal and Loratadine improve nose symptoms such as stuffy/blocked, sneezing, runny nose, itchy nose, sore nose and post-nasal drip in patients with AR. Symptoms fatigue, loss of taste, afraid of cold/wind and cold limb were improved significantly in moxibustion with Chinese herbal group. The mean quality of life scores decreased in both groups after treatment. Compare to control group, moxibustion with Chinese herbal is more effective than Loratadine in improving the quality of life in patients with AR. The results show moxibustion with Chinese herbal was effective to reduce symptoms and enhance quality of life in patients with allergic rhinitis. It is a simple, convenient and economic therapy for patients with AR. Further controlled trials of its effects in patients with allergic rhinitis are recommended.

[The effect of "Jian Pi Dao Zhi Fang" on human gastric carcinoma cells].

OBJECTIVE: To study the effect of Traditional Chinese medicine "Jian Pi Dao Zhi Fang" (JPDZF) with invigorating the spleen and purgative function on the growth of gastric carcinoma cells and the expression of VEGF and P53 in gastric carcinoma cells. METHODS: Gastric carcinoma cells were cultured in the RPMI-1640 media with 10% serum containing JPDZF for 48 hours. The cell cycle were tested with flow cytometer. Expression of P53, VEGF and its receptor Fit were evaluated by immunohistochemistry. RESULT: The cell cycle of gastric carcinoma cells cultured in the media with JPDZF were different from the control. The expression of P53, VEGF, Flt decreased with the dosage increasing of JPDZF. CONCLUSION: Traaditional Chinese medicine JPDZF can inhibit the growth of gastric carcinoma cells and the expression of P53, VEGF, Flt.