Objective:To investigate the role and underlying mechanisms of methyltransferase (Mettl) 3 in the process of angiotensin Ⅱ (Ang Ⅱ)-induced pericyte-to-myofibroblast transdifferentiation and renal fibrosis.Methods:C57BL/6J mice were used, in cell experiments, mouse renal pericytes were isolated and cultured using magnetic bead sorting. These pericytes were then induced to transdifferentiate into myofibroblasts with 1×10 6 mmol/L Ang Ⅱ, which was the Ang Ⅱ group, while pericytes cultured in normal conditions served as the control group. Successful transdifferentiation was verified by immunofluorescence staining, Western blotting, and real-time reverse transcription PCR (RT-qPCR) for α-smooth muscle actin (α-SMA). The levels of m6A modifications and related enzymes (Mettl3, Mettl14), Wilms tumor 1-associated protein (WTAP), fat mass and obesity protein (FTO), ALKBH5, YTHDF1, YTHDF2, YTHDC1, YTHDC2, YTHDC3 were assessed by Dot blot, RT-qPCR and Western blot. Mettl3 expression was inhibited in cells using lentivirus-mediated Mettl3-shRNA transfection, creating sh-Mettl3 and Ang Ⅱ+sh-Mettl3 groups, while lentivirus empty vector transfection served as the negative control (Ang Ⅱ+sh-NC group). The impact of Ang Ⅱ on pericyte transdifferentiation was observed, and the expression of downstream phosphatidylinositol 3-kinase (PI3K)/AKT signaling pathway proteins, including PI3K, AKT, phosphorylated AKT at serine 473 (p-AKT (S473)), and phosphorylated AKT at threonine 308 (p-AKT (T308)), were examined. PI3K gene transcription was inhibited by co-culturing cells with actinomycin D, and the half-life of PI3K mRNA was calculated by measuring residual PI3K mRNA expression over different co-culture time. The reversibility of Mettl3 inhibition on Ang Ⅱ-induced pericyte-to-myofibroblast transdifferentiation was assessed by adding the AKT activator SC79 to the Ang Ⅱ+sh-Mettl3 group. In animal experiments, mice were divided into these groups: sham group (administered 0.9% sterile saline), Ang Ⅱ group (infused with Ang Ⅱ solution), sh-Mettl3 group (injected with Mettl3 shRNA lentivirus solution), Ang Ⅱ+sh-Mettl3 group (infused with Ang Ⅱ solution and injected with Mettl3 shRNA lentivirus solution), and Ang Ⅱ+sh-Mettl3+SC79 group (administered Ang Ⅱ solution and Mettl3 shRNA lentivirus, with an additional injection of SC79). Each group consisted of six subject mice. Blood pressure was measured using the tail-cuff method before and after surgery, and serum creatinine, urea, and urinary albumin levels were determined 4 weeks post-surgery. Kidney tissues were collected at 28 days and stained using hematoxylin-eosin (HE) and Masson′s trichrome to assess the extent of renal fibrosis. Results:Primary renal pericytes were successfully obtained by magnetic bead sorting, and intervened with 1×10 6 mmol/L Ang Ⅱ for 48 hours to induce pericyte-to-myofibroblast transdifferentiation. Dot blot results indicated higher m6A modification levels in the Ang Ⅱ group compared to the control group ( P<0.05). RT-qPCR and Western blot results showed upregulation of Mettl3 mRNA and protein levels in the Ang Ⅱ group compared to the control group (both P<0.05). In the Ang Ⅱ+sh-Mettl3 group, Mettl3 protein expression was lower than that in the Ang Ⅱ group, with reduced expression levels of α-SMA, vimentin, desmin, fibroblast agonist protein (FAPa) and type Ⅰ collagen (all P<0.05). Compared to the control group, PI3K mRNA expression level was elevated in the Ang Ⅱ group, along with increased p-AKT (S473) and p-AKT (T308) expressions. In the Ang Ⅱ+sh-Mettl3 group, PI3K mRNA expression and p-AKT (S473) and p-AKT (T308) levels were decreased (all P<0.05). The half-life of PI3K mRNA was shorter in the Ang Ⅱ+sh-Mettl3 group than that in the Ang Ⅱ+sh-NC group (2.34 h vs. 3.42 h). The ameliorative effect of Mettl3 inhibition on Ang Ⅱ-induced pericyte-to-myofibroblast transdifferentiation was reversible by SC79. Animal experiments showed higher blood pressure, serum creatinine, urea, and 24-hour urinary protein levels, and a larger fibrosis area in the Ang Ⅱ group compared to the sham group (all P<0.05). The fibrosis area was smaller in the Ang Ⅱ+sh-Mettl3 group than that in the Ang Ⅱ group ( P<0.05), but increased again upon addition of SC79. Conclusion:Mettl3-mediated RNA m6A epigenetic regulation is involved in Ang Ⅱ-induced pericyte-to-myofibroblast transdifferentiation and renal fibrosis, potentially by affecting PI3K stability and regulating the PI3K/AKT signaling pathway.

Objective To analyze the epidemiological characteristics of syphilis in Yichang City from 2011 to 2020, and provide a scientific basis for developing effective control measures. Methods The data of syphilis cases reported in Yichang City from 2011 to 2020 were collected and analyzed by descriptive epidemiological methods. Results A total of 8 107 cases of syphilis were reported in Yichang City from 2011 to 2020, with an average annual reported incidence rate of 19.76 per 100 000.The reported incidence of syphilis increased from 17.39/100 000 in 2011 to 24.49/100 000 in 2018 （APC=6.82%，95%CI：2.34%~11.49%， P＜0.05）, from 24.49/100 000 in 2018 Decline to 17.55/100 000 in 2020（P＞0.05）. Recessive syphilis is the main type, with 5 837 cases (72.00%), and the annual reported incidence rate is 14.23 per 100 000. The number of cases peaked in July, at 836 (10.31%), and 433 (5.34%) of the lowest in February . The top 3 counties with annual average reported incidence rates were Zigui County (28.51/100 000) , Xiling District (25.01/100 000), and Dangyang City (23.73/100 000). There were a statistically significant differences in reported incidence between counties (2 =933.92, P＜0.01). Gender ratio was 0.72. Cases mainly concentrated in people over 20 years old, 7901 cases (97.46%). The occupation distribution is mainly farmers, housework, unemployed, and retired people. Conclusion The incidence of syphilis in Yichang City has been on the rise in recent ten years, and there is still a long way to go to prevent and control syphilis. We suggested to continue to strengthen the prevention and control of syphilis and comprehensively prevent and control the spread of syphilis.

OBJECTIVE To study the quality grade stand ard of the premature Forsythia suspensa . METHODS A total of 138 batches of premature F. suspensa were collected from the main producing areas of F. suspensa in China. According to 2020 edition of Chinese Pharmacopoeia ，the contents of impurities ，moisture，ethanol-soluble extract ，volatile oil ，forsythin and forsythoside A in the premature F. suspense were determined ，and the qualified samples were screened. AHP-PCA mixed weighting method was used to give comprehensive weight to the indicators （except for the limit of impurity ）. The comprehensive score of the samples was calculated. The suggestions on the quality grade division of premature F. suspensa were put forward according to cluster analysis of K-mean value. RESULTS & CONCLUSIONS The contents of impurities ，moisture，ethanol-soluble extract ，volatile oil ，forsythin and forsythoside A in the premature F. suspense were 0-7.80％，1.60％-8.18％，13.13％-61.60％，0.21％-3.47％，0.02％-2.15％ and 0.79％-14.04％，respectively；average contents of them were 1.24％，4.97％，34.88％，2.01％，0.42％，6.86％，respectively. Totally 47 batches of 138 batches were qualified in all indexes. It is suggested that the quality grade of the premature F. suspense can be divided into three grades ：in first grade of F. suspense ，the contents of volatile oil ，forsythin，forsythoside A ， ethanol-soluble extract and moisture were ≥2.40％，≥0.59％，≥8.34％，≥38.66％ and ≤4.99％，respectively；in second grade of F. suspense ，the contents of above indicators were ≥2.26％，≥0.41％，≥7.47％，≥32.58％ and ≤5.33％，respectively；in third grade of F. suspense ，the contents of above indicators were ≥2.15％，≥0.32％，≥4.60％，≥31.52％ and≤7.23％，respectively.

Objective:To explore the efficacy and safety of ticagrelor versus clopidogrel in acute coronary syndrome (ACS) Chinese patients using glycoprotein Ⅱb/Ⅲa inhibitor (GPI).Methods:The data from CCC-ACS (Improving Care for Cardiovascular Disease in China-ACS) project were systematically reviewed in ACS patients with GPI. The patients were divided into ticagrelor and clopidogrel groups. A logistic analysis and propensity score matching (PSM) were performed to compare occurrences of major cardiovascular events (MACE) and bleeding events between the two groups during hospitalization.Results:A total of 63 641 ACS patients were collected from 150 hospitals. Logistic regression analyses showed that there was no statistically significant difference in the reduction of MACE between ticagrelor and clopidogrel when using GPI ( OR=0.881, 95% CI 0.599-1.296; P=0.521). However, major bleeding rate was higher in the ticagrelor group than that in the clopidogrel group ( OR=1.401, 95% CI 1.075-1.852; P=0.013). Similar results were observed after PSM. No statistic difference in MACE between the ticagrelor and clopidogrel group ( OR=0.919, 95% CI 0.613-1.376; P=0.681). Major bleeding rate was higher in the ticagrelor group ( OR=1.559, 95% CI 1.130-2.150; P=0.007). Conclusion:In ACS patients with GPI, ticagrelor did not reduce MACE, but increased the major bleeding risk compared with clopidogrel.

Objective To analyze the changes of CD4⁺ T cell numbers at different periods of antiviral therapy in HIV-infected and AIDS patients (HIV/AIDS) in Yichang City. Methods The relevant information was retrieved from the National AIDS Comprehensive Prevention Information System-Antiretroviral Treatment Management Database. Changes in the number of CD4⁺ T cells were analyzed in HIV/AIDS patients who started receiving highly active antiretroviral therapy (HAART) and continued the treatment for 3 years from January 1, 2003 to December 31, 2017 in Yichang. Results The number of CD4⁺T lymphocytes in 550 HIV/AIDS cases increased significantly at various time points within 3 years after treatment, and increased with the increase of treatment time（F=100.20，P<0.001）. The CD4+T cell counts of different baseline level groups were statistically different before and after treatment（F=8.57，P<0.01）. The CD4+ T cell counts of patients who started treatment at age of 15-30 years old increased faster than those who started treatment at age of over 30 years old（F=1.27，P<0.05）. Conclusion HAART has a significant effect on the increase of CD4+T cells, and the early treatment is more effective. Early detection, diagnosis and treatment should be promoted, and anti-viral treatment should be actively carried out.

Objective:To evaluate the clinical effect of transferred direct popliteal cutaneous artery perforator propeller flap for repairing soft tissue defects of the popliteal fossa.Methods:The clinical data of 9 patients with defects of popliteal fossa from June 2013 to June 2019 were analyzed retrospectively, in which including 6 men and 3 women whose ages ranged from 25 to 73 years old. The sizes of soft tissue defects ranged from 6.5 cm×3.5 cm to 17.0 cm×8.5 cm. Perforator propeller flap was designed and cut to repair the defects according to the location, size and shape of defects, in which the largest was 18.0 cm×10.0 cm and the smallest was 7.5 cm×4.5 cm. To evaluate the effect according to survival, infection control, elasticity and color, appearance of the flaps, the scar at the donor site, cutaneous sense, knee joint function, and patients’satisfaction.Results:All the flaps survived. The incisions in donor and recipient site were primary healing. The survival of flaps was good. The appearance, color, and elasticity were close to normal. On the other hand, the scar at the donor site was small after 6 to 60 months follow-up. 2 cases appeared necrosis on edge of distal flaps and were healed after dressing. The efficacy was satisfactory in 8 cases, general in 1 case and without dissatisfactory. The function of knee joint was excellent in 7 cases, good in 1 case, and general in 1 case. Skin swelling rating in early-stage was 1st degree in 6 patients, 2nd degree in 2 patients, 3rd degree in 1 patient; in the later stage was 1st degree in 8 patients, 2nd degree in 1 patient.Conclusions:It is simple, safe and reliable to repair soft tissue defects of popliteal fossa with transferred direct popliteal cutaneous artery perforator propeller flap, which would be an ideal method for repairing soft tissue defects of the popliteal fossa.

Objective:To evaluate the clinical effect of transferred direct popliteal cutaneous artery perforator propeller flap for repairing soft tissue defects of the popliteal fossa.Methods:The clinical data of 9 patients with defects of popliteal fossa from June 2013 to June 2019 were analyzed retrospectively, in which including 6 men and 3 women whose ages ranged from 25 to 73 years old. The sizes of soft tissue defects ranged from 6.5 cm×3.5 cm to 17.0 cm×8.5 cm. Perforator propeller flap was designed and cut to repair the defects according to the location, size and shape of defects, in which the largest was 18.0 cm×10.0 cm and the smallest was 7.5 cm×4.5 cm. To evaluate the effect according to survival, infection control, elasticity and color, appearance of the flaps, the scar at the donor site, cutaneous sense, knee joint function, and patients’satisfaction.Results:All the flaps survived. The incisions in donor and recipient site were primary healing. The survival of flaps was good. The appearance, color, and elasticity were close to normal. On the other hand, the scar at the donor site was small after 6 to 60 months follow-up. 2 cases appeared necrosis on edge of distal flaps and were healed after dressing. The efficacy was satisfactory in 8 cases, general in 1 case and without dissatisfactory. The function of knee joint was excellent in 7 cases, good in 1 case, and general in 1 case. Skin swelling rating in early-stage was 1st degree in 6 patients, 2nd degree in 2 patients, 3rd degree in 1 patient; in the later stage was 1st degree in 8 patients, 2nd degree in 1 patient.Conclusions:It is simple, safe and reliable to repair soft tissue defects of popliteal fossa with transferred direct popliteal cutaneous artery perforator propeller flap, which would be an ideal method for repairing soft tissue defects of the popliteal fossa.

Objective:To evaluate the clinical effect of soft tissue defects around knee joints repaired with thigh distal perforator propeller flap.Methods:Clinical data of 72 patients with defects around knee joints from January, 2009 to October, 2019 were analyzed retrospectively, including 43 males and 29 females, aged from 17 to 83 years. The sizes of soft tissue defects ranged from 4.0 cm×3.0 cm to 22.0 cm×13.0 cm. Perforator propeller flaps were designed and harvested to repair the defects according to anatomical features of the origin, course, branches, distribution and anastomosis of vascular network of the perforator vessels and location as well as the size and shape of defects. Nine patients repaired with the lateral distal perforator propeller flaps (LDF), 53 with descending genicular artery perforator propeller flaps (DGF) and 10 with direct popliteal artery perforator propeller flaps (DPF). The size of flap was 4.5 cm×3.5 cm to 24.0 cm×14.0 cm. The donor sites were sutured directly or covered with free full-thickness skin. To evaluate the effect, the skin swelling rating in early and later stage were taken. Three to 72 months of followed-up was conducted through outpatient clinic (78%), telephone (15%) and WeChat (7%).Results:The flaps totally survived in 65 patients and 7 patients underwent partial necrosis at the distal end of the flaps ( 2 cases of LDF, 4 cases of DGF, 1 case of DPF) who were treated with dressing, and the donor sites were completely healed. The appearance and shape of the repaired knee joints was good, and the color and elasticity was close to surrounding normal skin, TPD of the flaps were 7-10 mm, and the scar was small after 3 to 72 months' follow-up. The efficacy was satisfactory in 48 cases ( 4 cases of LDF, 39 cases of DGF, 5 cases of DPF), general in 24 cases ( 5 cases of LDF, 14 cases of DGF, 5 cases of DPF). Degree of flap swelling: in early stage, I° in 40 cases ( 4 cases of LDF, 31 cases of DGF, 5 cases of DPF), II° in 20 cases ( 3 cases of LDF, 14 cases of DGF, 3 cases of DPF), III° in 10 cases ( 2 cases of LDF, 7 cases of DGF, 1 case of DPF), IV° in 2 cases(1 case of DGF, 1 case of DPF); in later stage: I° in 55 cases ( 6 cases of LDF, 41 cases of DGF, 8 cases of DPF), II° in 15 cases ( 2 cases of LDF, 12 cases of DGF, 1 case of DPF), III° in 2 cases ( 1 case of LDF, 1 case of DPF). The evaluation of knee joint function: excellent in 17 cases ( 2 cases of LDF, 12 cases of DGF, 3 cases of DPF), good in 35 cases( 4 cases of LDF, 27 cases of DGF, 4 cases of DPF), general in 20 cases( 3 cases of LDF, 14 cases of DGF, 3 cases of DPF). The good rate was about 73%. The range of motion and stability of the knee joint were basically normal and there was no pain found to affect study and life. All the patients were satisfied with the effect.Conclusion:Repairing soft tissue defect around knee joint with thigh distal perforator propeller flap is reliable and clinical applicable.

Objective:To evaluate the clinical effect of soft tissue defects around wrist joints repaired with forearm distal perforator propeller flap.Methods:The clinical data of 125 patients with defects of popliteal fossa from May 2008 to October 2019 were analyzed retrospectively, in which 71 men and 54 women were included aged from 16 to 87 years old. The sizes of soft tissue defect ranged from 3.5 cm×2.5 cm to 12.0 cm×6.0 cm. Perforator propeller flap was designed and cut to repair the defects according to their location, size and shape. 62 cases of distal radial artery perforator propeller flap, 48 cases of distal ulnar artery perforator propeller flap and 15 cases of distal posterior interosseous artery perforator propeller flap were included, in which the largest was 14.0 cm×6.0 cm and the smallest was 4.0 cm×3.0 cm. The donor sites were covered with free full-thickness skin grafts. Skin swelling rating in early and later stage and the color after 3 to 60 months follow-up were evaluated for the effect.Results:All the flaps survived in 116 patients, though the distal edge of the flaps were necrotic in 9 cases. The incisions in donor and recipient sites were primarily healed. After 3 to 60 months follow-up, we found no deformity of wrist joints and the appearance was good; the color and elasticity were close to normal; the scar was small; two-point discrimination was 6 mm to 9 mm. The outcome was satisfactory in 89 cases, average in 36 cases and without dissatisfactory. Skin swelling rating in early stage was first degree in 80 cases, second degree in 30 cases, third degree in 15 cases; in later stage was first degree in 85 cases, second degree in 35 cases, third degree in 15 cases. Wrist function was assessed in four aspects: palmar flexion, dorsiflexion, radial deviation, and ulnar deviation. Results were excellent in 29 cases, good in 63 cases, average in 33 cases and the good rate was 73.6%. The pain of wrist did not affect the patients’ daily routines and activities.Conclusions:It is reliable to repair soft tissue defects around wrist joints repaired with forearm distal perforator propeller flap which is worthy of clinical use, especially in middle-aged and senile patients.

Objective:To evaluate the clinical effect of soft tissue defects around wrist joints repaired with forearm distal perforator propeller flap.Methods:The clinical data of 125 patients with defects of popliteal fossa from May 2008 to October 2019 were analyzed retrospectively, in which 71 men and 54 women were included aged from 16 to 87 years old. The sizes of soft tissue defect ranged from 3.5 cm×2.5 cm to 12.0 cm×6.0 cm. Perforator propeller flap was designed and cut to repair the defects according to their location, size and shape. 62 cases of distal radial artery perforator propeller flap, 48 cases of distal ulnar artery perforator propeller flap and 15 cases of distal posterior interosseous artery perforator propeller flap were included, in which the largest was 14.0 cm×6.0 cm and the smallest was 4.0 cm×3.0 cm. The donor sites were covered with free full-thickness skin grafts. Skin swelling rating in early and later stage and the color after 3 to 60 months follow-up were evaluated for the effect.Results:All the flaps survived in 116 patients, though the distal edge of the flaps were necrotic in 9 cases. The incisions in donor and recipient sites were primarily healed. After 3 to 60 months follow-up, we found no deformity of wrist joints and the appearance was good; the color and elasticity were close to normal; the scar was small; two-point discrimination was 6 mm to 9 mm. The outcome was satisfactory in 89 cases, average in 36 cases and without dissatisfactory. Skin swelling rating in early stage was first degree in 80 cases, second degree in 30 cases, third degree in 15 cases; in later stage was first degree in 85 cases, second degree in 35 cases, third degree in 15 cases. Wrist function was assessed in four aspects: palmar flexion, dorsiflexion, radial deviation, and ulnar deviation. Results were excellent in 29 cases, good in 63 cases, average in 33 cases and the good rate was 73.6%. The pain of wrist did not affect the patients’ daily routines and activities.Conclusions:It is reliable to repair soft tissue defects around wrist joints repaired with forearm distal perforator propeller flap which is worthy of clinical use, especially in middle-aged and senile patients.