OBJECTIVES: To explore the mechanism by which Tougu Xiaotong Capsule (TXC) promotes chondrogenic differentiation and cartilage repair in mice with osteoarthritis (OA). METHODS: Fifty 8-week-old male C57BL mice were randomly divided into normal control group, cartilage damage (induced by subchondral ring-shaped drilling) model group and TXC treatment groups at low, moderate and high doses (184, 368 and 736 mg/kg, respectively). Saline (in normal control and model groups) and TXC were administered after modeling by daily gavage for 6 consecutive weeks. The changes of cartilage damage in the mice were assessed by measuring thermal withdrawal latency (TWL) and mechanical withdrawal threshold (MWT) and using micro-CT, modified safranine O and fast green staining, HE staining, and qPCR. Primary cultures of mouse synovial mesenchymal stem cells (SMSCs) with lentivirus vector transfection for interfering CXCL12, TXC treatment, or both for 24 h were examined for chondrogenic differentiation using immunofluorescence staining, scratch assay, immunocytochemistry, and Western blotting. RESULTS: In mouse models with cartilage damage, TXC treatment at the moderate dose significantly alleviated joint pain, promoted cartilage repair, and upregulated the mRNA expression levels of CXCL12, GDF5, collagen II, aggrecan, Comp and Sox9 in the cartilage tissue. In primary mouse SMSCs, CXCL12 knockdown resulted in significant reduction of GDF5 protein expression, migration ability and Sox9 protein expression, and these changes were obviously reversed by TXC treatment. CONCLUSIONS TXC promotes chondrogenic differentiation of mouse SMSCs to promote repair of cartilage damage in mice by activating the CXCL12/GDF5 pathway.
Animals ; Drugs, Chinese Herbal/therapeutic use* ; Osteoarthritis/metabolism* ; Male ; Growth Differentiation Factor 5/metabolism* ; Mice, Inbred C57BL ; Mice ; Chemokine CXCL12/metabolism* ; Signal Transduction/drug effects* ; Cell Differentiation/drug effects* ; Cartilage, Articular/drug effects* ; Mesenchymal Stem Cells/cytology*

Objective:To explore the repair methods of complex facial defect wounds involving paranasal sinuses and their clinical effectiveness.Methods:A retrospective observational study was conducted. From January 2020 to May 2022, 5 patients admitted to the Department of Burns and Plastic Surgery of Xiangya Hospital of Central South University and 4 patients admitted to the Department of Burns and Plastic Surgery of Chenzhou First People's Hospital with complex facial defect wounds involving paranasal sinuses met the inclusion criteria, including 6 males and 3 females, aged 35-69 years, including 4 patients with titanium mesh exposure combined with paranasal sinuses injury and 5 patients with tumor involving paranasal sinuses. After an adequate assessment of the damage by a multiple discipline team, titanium mesh removal, paranasal sinus debridement, and paranasal sinus mucosa removal were performed in patients with exposed titanium mesh, and radical tumor resection was performed in patients with tumors, with postoperative skin and soft tissue defects areas of 5.0 cm×2.5 cm to 18.0 cm×7.0 cm, anterior paranasal sinus wall defects/absence areas of 3 cm×2 cm to 6 cm×4 cm, and sinus cavity depths of 1 to 4 cm. Depending on the perforator course of the descending branch of the lateral circumflex femoral artery, the anterolateral femoral chimeric flap or anterolateral femoral myocutaneous flap (with flap area of 9 cm×4 cm to 19 cm×8 cm, muscle size of 5 cm×3 cm×3 cm to 11 cm×6 cm×3 cm) was transplanted to repair the defect, and the donor site wound was sutured directly. The type of tissue flap transplanted, the blood vessel of the recipient area, and the vascular anastomosis way during the operation, the recovery of the donor and recipient areas and the occurrence of complications after operation were observed. The appearance and blood supply of the recipient area and the recurrence of ulcers and tumors were followed up.Results:The anterolateral femoral myocutaneous flap transplantation was performed in 6 patients, and the anterolateral femoral chimeric flap transplantation was performed in 3 patients. The blood vessels in recipient areas were facial arteries and veins in 3 cases and superficial temporal arteries and veins in 6 cases. The superficial temporal arteries and veins were bridged with blood vessels in tissue flaps by flow-through way in 2 patients, and end-to-end anastomosis of blood vessels in donor and recipient areas was performed in 7 patients. After operation, all the tissue flaps survived, and the facial defect wounds were well repaired without cerebrospinal fluid leakage or paranasal sinus secretion leakage, no intracranial infection occurred, and the wounds in donor areas were healed well. Follow-up of 6-35 months after operation showed that all the patients had good blood supply in the recipient area, and the shape was acceptable; 4 patients with exposed titanium mesh had no recurrence of ulceration, and 5 patients with tumor had no local tumor recurrence or metastasis.Conclusions:Based on an adequate assessment of the extent of paranasal sinuses involved in the facial wound and the nature of the defect, good clinical effects can be achieved by using the anterolateral femoral muscle flap or the anterolateral femoral chimeric flap transplantation to repair complex facial defect wounds with open paranasal sinuses.

Objective:To evaluate the safety and efficacy of combination of transjugular intrahepatic portosystemic shunt (TIPS) combined with interventional therapy for primary hepatic carcinoma complicated with portal hypertension.Methods:From October 2013 to December 2017, medical records of 141 patients with primary hepatic carcinoma complicated with portal hypertension hospitalized and treated in Anhui Provincial Cancer Hospital were collected. According to the inclusion and exclusion criteria, 32 cases were in the TIPS combined with interventional treatment group and 29 cases were included in the intervention-only group after age, gender, mean tumor diameter and Child-Pugh classification matched with TIPS combined with intervention treatment group. The efficacy of TIPS was obsented, and the related complications and prognosis of the two groups at six, 12 and 24 months after treatment were compared. Independent sample t test, Mann-Whitney U test and Chi-square test were used for statistical analysis, and Kaplan-Merier method was used for survival analysis. Results:There were no statistically significant differences between TIPS combined with intervention group and intervention-only group in age, gender, mean tumor diameter, Child-Pugh classification, Child-Turcotte-Pugh (CTP) score, Barcelona staging, interventional treatment, causes of liver cirrhosis, portal hypertension related symptoms , portal vein tumor thrombus, alanine aminotransferase (ALT), and total bilirubin (TBil) (all P>0.05). The success rate of TIPS of patients in TIPS combined with intervention group was 100% (32/32). All the portal hypertension related symptoms after TIPS were relieved, and the remission rate was 100% (32/32). Compared with that before TIPS, after TIPS, the portal vein pressure decreased ((36.5±6.9) cmH 2O vs. (25.5±5.6) cmH 2O (1 cmH 2O=0.098 kPa)), the diameter of portal vein decreased ((15.9±3.5) mm vs. (13.7±2.7) mm), and ascites volume decreased (abdominal circumference ((105.6±13.9) cm vs. (88.0±9.8) cm), red blood cell count ((2.6±0.8)×10 12/L vs. (3.3±1.3)×10 12 /L) and hemoglobin level ((78.9±15.5) g/L vs. (108.4±14.6) g/L) both increased, and the differences were statistically significant ( t=2.866, 3.105, 10.769, -3.548 and -3.869, all P<0.01). The stent patency rate six months after TIPS was 100% (32/32), the 12-month patency rate was 94% (30/32), and the 24-month patency rate was 84% (27/32). Six months, 12 months and 24 months after treatment, the incidence of ascites of patients in the TIPS combined with interventional treatment group was 0, 3.1% (1/32) and 9.4% (3/32), respectively, and the incidence of bleeding was 3.1% (1/32), 9.4% (3/32) and 15. 6% (5/32), respectively, which were all lower than those of the intervention-only group (13.8%, 4/29; 27.6%, 8/29; 65.5%, 19/29; 20.7%, 6/29; 34.5%, 10/29 and 62.1%, 18/29), and the differences were statistically significant ( χ2=4.72, 7.24, 20.80; and 4.62, 5.72, 13.97; all P<0.05). There were no statistically significant differences in the incidence rates of hepatic encephalopathy 6 and 12 months after treatment between the two groups (both P>0.05). The incidence of hepatic encephalopathy of intervention-only group (48.3%, 14/29) 24 months after treatment was higher than that of TIPS combined with intervention group (21.9%, 7/32), and the difference was statistically significant ( χ2=4.70, P=0.030). The results of Kaplan-Merier analysis indicated that 12 and 24 months after treatment the cumulative survival rates of TIPS combined with intervention group (84.4%, 27/32 and 53.1%, 17/32) were both higher than those of the intervention-only group (41.4%, 12/29 and 13.8%, 4/29), and the differences were statistically significant ( χ2=12.20 and 5.06, both P<0.05). The median survival time of TIPS combined with intervention group was 25 months, which was longer than that of the intervention-only group (12 months), and the difference was statistically significant ( Z=3.341, P=0.001). Conclusions:TIPS combined with interventional therapy is safe and effective in the treatment of primary hepatic carcinoma complicated with portal hypertension, which can improve the quality of life and increase the survival rate.

OBJECTIVE: To apply single cell sequencing based on multiple annealing and looping amplification cycles (MALBAC) for the determination of the rate and type of mosaicisms of high-quality embryos at cleavage stage. METHODS: After thawing and removing of zona pellucida by enzymatic digestion, blastomeres were collected the high-quality embryos donated by couples whom had given birth to healthy offspring by intracytoplasmic sperm injection and embryo transfer. The whole genome of single cell was amplified and subjected to next generation sequencing. RESULTS: From a total of 23 embryos, 184 blastomeres were collected. 175 (95.1%) of the blastomeres were successfully sequenced, of which 100 (57.1%) were found to harbor chromosomal aneuploidies. Among the 23 embryos, 3 (13.0%) were diploid, 20 (87.0%) were mosaicisms, which included 5 (21.7%) aneuploid mosaicisms, 7 (30.4%) diploid-aneuploid mosaicisms, 5 (21.7%) abnormal mosaicisms, and 3 (13.0%) irregular segregations. CONCLUSION There is a high rate of chromosomal mosaicisms in high-quality cleavage embryos. Mosaicisms of complex chromosomal abnormality or with high proportion of abnormal cells may be an important factor affecting the potential of embryonic development.

Objective:To compare the clinical outcomes of one and two blastocysts in the freeze-thaw transplantation cycle.Methods:Totally 3 675 cycles of frozen thawed blastocyst transplantation in Reproductive Medical Center of the Second Nanning People′s Hospital from January 2012 to December 2016 were analyzed retrospectively. According to the quantity and quality of transferred blastocysts, all the patient were divided into two groups: (1) one embryo group, including the single excellent group (one high quality blastocyst) and the single non excellent group (one non high quality blastocyst); (2) two embryo groups, including the double excellent group (two high quality blastocysts), the one excellent and one non excellent group (one high quality blastocyst+one non high quality blastocyst), and the two non excellent group (two non high quality blastocysts were transplanted). Then the patients were divided into subgroups according to their ages: less than 35 years old, 35-40 years old and over 40 years old. On this basis, the implantation rate, clinical pregnancy rate, multiple birth rate and live birth rate were compared.Results:(1) The implantation rate, clinical pregnancy rate, multiple birth rate, preterm birth rate and live birth rate were all significantly increased, while the abortion rate was significantly reduced in the double blastocyst group (all P<0.05). (2) In the group of<35 years old, the rates of multiple birth and preterm birth in the double blastocyst group were significantly higher than those in the single optimal group ( P<0.01). (3) In the 35-40 years old group, the clinical pregnancy rate, multiple birth rate and live birth rate of the double excellent group were significantly higher than those of the single excellent group ( P<0.01); while the clinical pregnancy rate and live birth rate of the one excellent and one non excellent group and the double non excellent group were not significantly different from those of the single excellent group ( P>0.05), but the multiple birth rate and preterm birth rate were significantly increased ( P<0.01). The clinical pregnancy rate, live birth rate and multiple birth rate of double non optimal group were significantly higher than those of single non optimal group ( P<0.01). (4) In the group>40 years old, there were no significant differences in clinical pregnancy rate and live birth rate between the two groups ( P>0.05). There were no significant differences in implantation rate, clinical pregnancy rate and live birth rate between double non optimal group and single non optimal group ( P>0.05). Conclusion:No matter the age of the patients, if the couple have high quality blastocysts, we should give priority to single high quality blastocyst transplantation; even if they have no high quality blastocysts, we should also consider single blastocyst transplantation, in order to reduce the risk of multiple pregnancy and improve the cumulative live birth rate, so as to improve the pregnancy outcome.

Objective To evaluate the efficacy of transjugular intrahepatic portosystemic shunt (TIPS)in the treatment of patients with hepatic sinusoidal obstruction syndrome (HSOS).Methods From April 2015 to August 2018,at The First Affiliated Hospital of University of Science and Technology of China,21 patients with gynura segetum caused HSOS were selected.All the patients received TIPS treatment because of unresponsiveness to anticoagulant therapy for at least two weeks.After operation patients were followed up with liver and portal vein Doppler ultrasonography examination,liver and kidney function tests,and survival observation.T test,logistic univariate regression analysis and Cox regression analysis were performed for statistical analysis.Results Among the 21 patients with gynura segetum-related HSOS,18 patients were in the subacute phase and three patients in the chronic phase.All of them were moderate or severe patients and all successfully underwent TIPS.The postoperative portal vein pressure was (16.71 ± 4.68) cmH2O (1 cmH2O =0.098 kPa),which was lower than that before operation ((41.52 ±6.27) cmH2O),and the difference was statistically significant (t =16.936,P ＜ 0.01).The postoperation portal vein blood flow velocity was (41.52 ±7.70) cm/s,which was higher than before operation ((11.19 ± 3.29) cm/s),and the difference was statistically significant (t =-15.191,P ＜0.01).At one month after operation,15 of 21 patients were clinically cured;among the remaining six patients,four patients were improved and two patients were ineffective (including one patient died).At four months after operation,two patients died,and the remaining 19 patients were clinically cured.At one month after operation,the levels of alanine aminotransferase (ALT),aspartate aminotransferase (AST),total bilirubin (TBil) and serum creatinine were (23.7 ± 16.8) U/L,(33.9 ±7.4) U/L,(52.7 ± 38.2) μmol/L and (62.7 ± 12.6) μmol/L,respectively,which were lower than those before operation ((60.5 ± 42.4) U/L,(78.4 ± 42.4) U/L,(74.9 ± 38.2) μmol/L and (82.4 ± 19.6) μmol/L,respectively),and the differences were statistically significant (t =3.193,3.493,2.378 and 4.519;all P ＜ 0.05).The level of albumin was (39.0 ±3.1) g/L,which was higher than that before operation ((30.9 ± 3.8) g/L),and the difference was statistically significant (t =-10.283,P ＜ 0.01).Portal vein thrombosis and preoperative TBil level had predictive value for therapeutic efficacy (both P ＜0.05).The one-year cumulative survival rate of patients was 90.5％.Preoperative TBil level and hepatic encephalopathy had effects on the prognosis of patients (both P ＜ 0.05).Conclusion TIPS is a safe,reliable and effective treatment for patients with subacute and chronic gynura segetum-related HSOS who are not responding to ineffective anticoagulant therapy,which can improve the prognosis and survival rate of the patients.

Objective To explore the relationship of Crohn's disease (CD) susceptibility to aryl hydrocarbon receptor (AhR) polymorphisms and haplotypes in Han population in Wenzhou city, China. Methods A total of 310 CD patients and 573 age-and sex-matched healthy controls were enrolled in our study. Three single nucleotide polymorphisms (SNPs) of AhR(rs10249788,rs2158041,rs2066853) were determined by the improved multiple ligase detection reaction technique. Unconditional logistic regression analyses was applied to analyze the allelic and genotypic differences of each SNP between CD patients and controls, as well as their influence on the clinicopathologic characteristics in CD patients. Analyses of linkage disequilibrium and haplotype were performed by Haploview 4.2 software in all study subjects. Results Compared with the controls, the variant allele (T) and genotype (CT+TT) of (rs2158041) were evidently decreased among CD patients (19.52% vs. 25.04%, P=0.009; 34.19% vs. 44.68%, P=0.003). According to"the Montreal Classification Standards", CD patients were divided into different subgroups. The variant allele(T)and genotype(CT+TT)of(rs2158041)were significantly lower in patients with terminal ileum CD than in controls (16.79% vs. 25.04%, P=0.005; 28.24% vs. 44.68%, P=0.001). Similar conclusions were also drawn in patients with constricting disease when compared with the controls(15.20%vs.25.04%,P=0.003;28.43% vs.44.68%,P=0.003).The three SNPs above were shown to be in a linkage disequilibrium.Compared with the controls respectively,the frequency of haplotype(CCG)was increased in CD patients (44.73% vs. 39.60%, P=0.039), whereas that of haplotype (CTG) was decreased (18.02% vs. 22.78%, P=0.047). Conclusions AhR (rs2158041) variation might influence the risk as well as the location and behavior of CD. The haplotype (CCG) possibly increase the risk of CD development, whereas haplotype(CTG)might decrease it.

Objective To explore the relationship between ulcerative colitis (UC) susceptibility and tumor necrosis factor superfamily member (TNFSF) 15 gene polymorphisms and haplotypes in Han nationality in Zhejiang province of China. Methods A total of 408 UC patients and 574 healthy controls were recruited in this study. Three single nucleotide polymorphisms of TNFSF15 (rs3810936, rs4263839, rs4979462) were examined by improved multiple ligase detection reaction (iMLDR) technique. Analyses of linkage disequilibrium (LD) and haplotype were performed by Haploview 4.2 software in all study subjects. Results The variant allele A and genotype (GA+AA) of rs4263839 were less frequent in UC patients than in controls (45.34% vs. 50.17%, P=0.035; 68.38% vs. 76.66%, P=0.004). According to the severity and location of disease, UC patients were divided into different subgroups. After multiple comparison correction (α=0.012 5), the frequencies of variant allele A and genotype (GA+AA) of rs4263839 were lower in patients with severe UC than in the controls (37.69% vs. 50.17%, P=0.007;60.00% vs. 76.66%, P=0.004). Similar findings were also drawn for patients with extensive colitis in contrast with the controls (42.22% vs. 50.17%, P=0.009; 63.33% vs. 76.66%, P<0.001). Furthermore, the haplotype analysis indicated that three SNPs above were in a strong LD. The frequency of haplotype TAC was lower in UC patients than in the controls (40.83% vs. 46.04%, P=0.023). Also it was less prevalent in patients with severe UC and patients with extensive colitis when compared with controls respectively (33.38% vs. 46.04%, P=0.005;37.22% vs. 46.04%, P=0.003). Conclusions TNFSF15 (rs4263839) variation might not only reduce the risk of UC, but also affect the severity and lesion location of UC. The haplotype TAC formed by rs3810936, rs4263839 and rs4979462 might be related to a lower risk of UC, especially in patients with severe colitis or patients with extensive colitis.

Objective: To investigate the key factor(s) of hepatitis B virus X protein (HBx) promoting B7-H6 gene activation. Methods: The DNA fragments of the B7-H6 promoter were amplified from the human genomic DNA using polymerase chain reaction(PCR). Products of PCR were digested by KpnI and HindⅢ, and inserted into luciferase reporter vector (pGL3-Basic). The correctness of the recombinant plasmid pGL3-B7-H6 was confirmed by sequencing. Human hepatoma cell line HepG2 were co-transfected with pGL3-B7-H6 and the eukaryotic expression vectors (pCMV-HBs、pCMV-HBc and pCMV-HBx), and the relative luciferase activity was detected.The different dose of HBx expression plasmids were transfected into the HepG2 cells, and the expression of B7-H6 protein were determined by Western blot. Results: A period of 2.2 Kb of B7-H6 promoter region were successfully cloned into pGL3-Basic, which was confirmed by DNA sequencing. The relative luciferase activity was significantly higher in the cells transfected with pGL3-B7-H6 than that in the cells transfected with the empty vector pGL3-Basic (5.24±1.25 vs. 1.12±0.31, P=0.005). The relative luciferase activity in HepG2 cells co-transfected with pGL3-B7-H6 and pCMV-HBx was remarkably increased compared with the control group (17.60±3.36 vs. 6.73±1.36, P=0.001). Western blot further demonstrated that HBx protein can significantly enhance B7-H6 protein expression. Conclusions Hepatitis B Virus X proteins enhanced B7-H6 promoter activity and promoted B7-H6 protein expression.

BACKGROUND:Previous studies have found that electroacupuncture can delay articular cartilage degeneration mediated by JAK-STAT signaling pathway through upregulating the expression level of transforming growth factor β1 as well as mRNA expression levels of STAT3, Smad3 and LepR. In the meanwhile, electroacupuncture can inhibit the mRNA expression of p38 and Fas mRNA mediated by MAPK signaling pathways, further inhibiting the apoptosis of chondrocytes. OBJECTIVE: To explore the effect of electroacupuncture on the degeneration of articular cartilage in rats with knee osteoarthritis based on Ras-Raf-MEK1/2-ERK1/2 signaling pathway. METHODS:120 male healthy Sprague-Dawley rats aged 2 months olds were selected and randomly divided into normal, model, 15-minite electroacupuncture and 30-minute electroacupuncture groups (n=30 per group). The rats in the latter three groups received the intra-articular injection of 4% papain bilaterally, and the remaining rats received no intervention. At 2 weeks after modeling, the latter two groups were respectively given 15- and 30-minute electroacupuncture, five times weekly for consecutive 12 weeks. The morphology of the cartilage was observed by hematoxylin-eosin staining, the expression level of interleukin-1β in the synovium was detected by ELISA assay, and the protein expression levels of Ras, Raf, MEK1/2, ERK1/2, C-MYC, C-FOS, and C-JUN were detected by western blot analysis. RESULTS AND CONCLUSION: Hematoxylin-eosin staining showed that: in the model group, the cartilage surface was rough, the cartilage layer became thinner, and the cartilage structure was damaged with incomplete tidal line; in the 15- and 30-minute electroacupuncture groups, the cartilage structure was complete with clear layers and complete tidal line. ELISA showed that the expression level of interleukin-1β in the model group was significantly higher than that in the normal group (P< 0.01), and the level in the 15- and 30-minute electroacupuncture groups was significantly lower than that in the model group (P < 0.05). Western blot assay found that compared with the normal group, the protein expression levels of Ras, Raf, MEK1/2, ERK1/2, C-MYC, C-FOS, and C-JUN were increased in the model group. However, all above protein levels except ERK1/2 in the 15- and 30-minute electroacupuncture groups were significantly lower than those in the model group (P < 0.01,P < 0.05). To conclude, electroacupuncture inhibits the degeneration of articular cartilage in osteoarthritisvia Ras-Raf-MEK1/2-ERK1/2 signaling pathway and downregulating the expression level of interleukin-1β.