Application of live biotherapeutic products and perspective in the treatment of inherited metabolic disease / 生物工程学报

Live biotherapeutic products (LBPs) refer to the living bacteria derived from human body intestinal gut or in nature that can be used to treat the human disease. However, the naturally screened living bacteria have some disadvantages, such as deficient therapeutic effect and great divergence, which fall short of the personalized diagnosis and treatment needs. In recent years, with the development of synthetic biology, researchers have designed and constructed several engineered strains that can respond to external complex environmental signals, which speeded up the process of development and application of LBPs. Recombinant LBPs modified by gene editing can have therapeutic effect on specific diseases. Inherited metabolic disease is a type of disease that causes a series of clinical symptoms due to the genetic defect of some enzymes in the body, which may cause abnormal metabolism the corresponding metabolites. Therefore, the use of synthetic biology to design LBPs targeting specific defective enzymes will be promising for the treatment of inherited metabolic defects in the future. This review summarizes the clinic applications of LBPs and its potential for the treatment of inherited metabolic defects.

Role of mitofusion 2 in high glucose-induced endoplasmic reticulum stress and apoptosis of podocytes / 中华肾脏病杂志

Objective:To investigate the role of mitofusion 2 (Mfn2) in high glucose (HG)-induced endoplasmic reticulum stress (ERS) and apoptosis of podocytes.Methods:(1) Streptozocin was used to induce a diabetes mellitus (DM) rat model. Renal histopathological changes in rats were observed by HE staining. Expression of Mfn2 and CCAAT/enhancer-binding protein homologous protein (CHOP) in glomeruli was observed by immunohistochemistry. Protein levels of Mfn2, protein kinase RNA-like ER kinase (PERK), phospho(p)-PERK, and CHOP in glomeruli were analyzed by Western blotting. (2) Conditionally immortalized human podocytes (HPC) cultured in vitro were divided into control, mannitol (MA) and HG groups. Expression of Mfn2 was observed by immunofluorescence. Protein levels of Mfn2, p-PERK, PERK and CHOP in HPC were analyzed by Western blotting. Podocyte apoptosis in each group was evaluated by flow cytometry with AnnexinⅤ-PE/7AAD double staining method. (3) HPC were divided into control, HG, HG+Mfn2-Myc plasmid-transfected and HG+control plasmid-transfected groups. Protein levels of Mfn2, p-PERK, PERK and CHOP in HPC were analyzed by Western blotting. Expression of CHOP was observed by immunofluorescence. Mitochondrial membrane potential in each group was observed by mitochondrial membrane potential assay kit with JC-1. Podocyte apoptosis in each group was evaluated by flow cytometry with AnnexinⅤ-PE/7AAD double staining method. Results:(1) Compared with the control group, the glomerular mesangial matrix of the DM group rats was significantly proliferated, and the expression of Mfn2 was down-regulated with the expression of ERS-related proteins p-PERK/PERK and CHOP up-regulated (all P<0.05). (2) Compared with the control group, Mfn2 was down-regulated and p-PERK/PERK and CHOP were up-regulated in HPC of HG group (all P<0.05). Apoptosis of HPC was also increased in HG group. There was no significant difference in the above indicators between the control group and the mannitol group (all P>0.05). (3) Compared with the HG group, mitochondrial membrane potential of HPC was alleviated and apoptosis of HPC was decreased in HG+Mfn2-Myc plasmid-transfected group ( P<0.05). P-PERK/PERK and CHOP were down-regulated in HG+Mfn2-Myc plasmid-transfected group (both P<0.05). There was no significant difference in the above indicators between the HG group and the HG+control plasmid-transfected group (all P>0.05). Conclusions:Mfn2 down-regulation in HG-stimulated podocytes may induce ERS to increase apoptosis of podocytes. Up-regulation of Mfn2 can alleviate the HG-induced ERS and apoptosis in podocytes.

Efficacy of hydroxychloroquine in patients with COVID-19: results of a randomized clinical trial

AimsStudies have indicated that chloroquine (CQ) shows antagonism against COVID-19 in vitro. However, evidence regarding its effects in patients is limited. This study aims to evaluate the efficacy of hydroxychloroquine (HCQ) in the treatment of patients with COVID-19. Main methodsFrom February 4 to February 28, 2020, 62 patients suffering from COVID-19 were diagnosed and admitted to Renmin Hospital of Wuhan University. All participants were randomized in a parallel-group trial, 31 patients were assigned to receive an additional 5-day HCQ (400 mg/d) treatment, Time to clinical recovery (TTCR), clinical characteristics, and radiological results were assessed at baseline and 5 days after treatment to evaluate the effect of HCQ. Key findingsFor the 62 COVID-19 patients, 46.8% (29 of 62) were male and 53.2% (33 of 62) were female, the mean age was 44.7 (15.3) years. No difference in the age and sex distribution between the control group and the HCQ group. But for TTCR, the body temperature recovery time and the cough remission time were significantly shortened in the HCQ treatment group. Besides, a larger proportion of patients with improved pneumonia in the HCQ treatment group (80.6%, 25 of 31) compared with the control group (54.8%, 17 of 31). Notably, all 4 patients progressed to severe illness that occurred in the control group. However, there were 2 patients with mild adverse reactions in the HCQ treatment group. Significance: Among patients with COVID-19, the use of HCQ could significantly shorten TTCR and promote the absorption of pneumonia. SignificanceAmong patients with COVID-19, the use of HCQ could significantly shorten TTCR and promote the absorption of pneumonia. Trial registrationURL: https://www.clinicaltrials.gov/. The unique identifier: ChiCTR2000029559.

Caution: The clinical characteristics of COVID-19 patients at admission are changing

BackgroundWith the emergence of 4rd generation transmission, the prevention and treatment of the novel coronavirus disease 2019 (COVID-19) has entered a new period. We aimed to report several changes in the clinical characteristics at admission of patients with COVID-19. MethodsClinical records and laboratory results of patients suffering from COVID-19 were retrospectively reviewed and matched with the admission dates to analyze the changes in characteristics at the onset of illness. ResultsOf the 89 affected patients, 31 [34.8%] patients were admitted from January 16 to 22, and 58 [65.2%] were admitted from January 23 to 29. Patients were admitted with more systemic symptoms, such as fever (21 [67.7%] of 31), fatigue (13 [41.9%] of 31), and myalgia (7 [22.6%] of 31), before January 23. More patients (10 [32.3%] of 31) admitted before January 23 had a small amount of sputum production compared with a smaller proportion (4 [6.9%] of 58) of the patients admitted after January 23. Other symptoms, such as cough, nausea, diarrhea, and chest tightness, were not significantly different between the two groups. In addition, the group admitted before January 23 had a larger proportion of patients with reduced lymphocyte (13 [54.2%] of 24), CD3 (11 [54.4%] of 21), and CD8 (9 [42.9%] of 21) counts and elevated serum amyloid A (SAA, 18 [75%] of 24). ConclusionsThe initial symptoms of recently infected patients seem more insidious, indicating that the new coronavirus may gradually evolve into a virus similar to influenza and latent in asymptomatic carriers for a long time. SummaryCompared with the cases admitted earlier, more hidden initial symptoms and improved immune system disorders appeared in COVID-19 patients infected recently.

Roles of AKAP1 in high-glucose induced mitochondrial fission in podocytes / 中华肾脏病杂志

Objective To investigate the roles of A kinase anchoring protein1(AKAP1)in high-glucose induced mitochondrial fission in podocytes.Methods Conditionally immortalized human podocytes were cultured in serum-free medium for 24 hours,and then exposed to different glucose concentration conditions in different time periods.The protein expressions of AKAP1 were observed by immunofluorescence,and AKAP1,dynamin related protein1 (Drp1) and phospho Ser 637-Drp1 (p-Drp1)were analyzed by Western blotting.AKAP1 siRNA was transfected to block AKAP1 expression.Podocytes were then divided into normal control group (5 mmol/L glucose),hypertonic group (30 mmol/L mannitol+5 mmol/L glucose),high glucose group (35 mmol/L glucose),and high glucose+AKAP1 siRNA group.Mitochondrial morphological changes were assessed by mitotracker red staining.Podocyte apoptosis was assessed by flow cytometry.Results Compared with normal group,high-glucose induced more podocytes apoptosis (P ＜ 0.05),more mitochondrial fission with decreased aspect ratio and form factor (all P ＜ 0.05).Upregulated AKAP1 protein level,and increased ratio of p-Drp1/Drp1 (all P ＜ 0.05) in time and concentration dependent manners were also obscrvcd.Compared with high glucose group,transfection of AKAP1 siRNA showed less apoptosis (P ＜ 0.05),less mitochondrial fission with increased aspect ratio and form factor (all P ＜ 0.05),and down-regulated AKAP1 protein level as well as p-Drp1/Drp1 ratio (all P ＜ 0.05).Conclusion High glucose induced mitochondrial fission might be induced through AKAP1-Drp1 pathway.

The orbicularis muscle partial resection combined with orbital fat flap or orbicularis muscle flap graft for treatment of crow′s feet in women / 中华整形外科杂志

Objective@#To observe the effect of the outer orbicularis muscle partial resection combined with orbital fat flap or orbicularis muscle flap transfer to block orbicularis muscle for improvement of crow′s feet.@*Methods@#From October 2010 to May 2015, 82 cases of females (aged from 27 to 54 years old) with crow′s feet were treated with new method(n=41) and traditional method(control, n=41). The new method included partial resection and orbital fat flap or orbicularis muscle flap transfer to block orbicularis muscle. The traditional method was performed to lift the skin and orbicularis oculi muscle to improve the crow′s feet. The operation time, the postoperative recovery time, eyes closing function and hematoma, nerve injury and other complications were recorded. 1, 3, 6 and 12 months after operation, the effect was assessed by one cosmetic surgeon who is unkown the groups. Completely correction was assed as 5 points and no improvement or even worse as 0 point. 12 months after operation, the patients satisfactory rate was recorded.@*Results@#75 cases were followed up, including 39 in new method group and 36 in control group. The operative time and postoperative recovery time were (50±5) min, (58±4) min and (5±1) d, (6.0±1.5) d in the new method group and control group respectively (P<0.05). The two groups both had good eyes closing function, no eyes closing fatigue or difficult or other serious complications such as hematoma, nerve injury. Visual analog scale were 3.2±0.6, 3.5±0.5, 4.2±0.8, 4.3±0.8 and 3.1±0.6, 3.4±0.9, 3.0±0.6, 2.8±0.5 in the new method group and control group at 1, 3, 6, 12 months after surgery respectively . Obviously, the effect in new method group was much better than that in control group after 6 and 12 months with statistical significance (P<0.05). 12 months after surgery, the satisfaction surveys showed satisfactory in 33 cases; fairly satisfactory in 4 cases; dissatisfactory in 2 cases in new method group, and satisfactory in 10 cases; fairly satisfactory in 15 cases; dissatisfactory in 11 cases in the control group, showing significant difference between the 2 groups (P<0.05).@*Conclusions@#The outer orbital orbicularis muscle partial resection combined with orbital fat flap or orbicularis muscle flap is reasonable and reliable to correct crow′s feet with short recovery and less complication. It is a worthy new method for recommendation.

Role of JLP on the epithelial to mesenchymal transition in renal tubular epithelial cells / 中华肾脏病杂志

Objective To observe the effect of JLP on transdifferentiation of human renal proximal tubular epithelial cells (HK-2),and to investigate the role of p38 MAPK signaling pathway in this process.Methods The knock-down plasmids of JLP were constructed.HK-2 cells were randomly divided into four groups:negative control cells (Ctrl-shRNA group),knock-down jlp cells (jlpshRNA group),negative control cells with FGF-2 treatment (FGF-2 group) and knock-down jlp cells with FGF-2 treatment(jlp-shRNA +FGF-2 group).The expressions of JLP,E-cadherin,TGF-β1,α-SMA,p-p38 MAPK protein were detected by Western blotting.After the induction of FGF-2 for 24 hours,the expressions of α-SMA,COL-Ⅰ,FN were detected by immunocytochemistry.Results Compared with Ctrl-shRNA group,the expression of JLP protein was significantly down-regulated in FGF-2 group.Compared with FGF-2 group,the expressions of TGF-β1,α-SMA,p-p38 MAPK protein were significantly up-regulated,while E-cadherin protein was significantly down-regulated (P ＜ 0.05).Compared with FGF-2 group,the expressions of α-SMA,COL-Ⅰ,FN immunostaining increased markedly in jlp-shRNA+FGF-2 group.Conclusion Scaffolding protein JLP is critical in preventing EMT in the course of fibrosis through the inhibition of p-p38 activation in HK-2 cells.

Observations on the Efficacy of Electroacupuncture plus Turgunmed Training System in Treating Lumbar Intervertebral Disc Herniation / 上海针灸杂志

Objective To compare the clinical efficacies of electroacupuncture plus rehabilitation training versus electroacupuncture alone in treating lumbar intervertebral disc herniation and evaluate the therapeutic effect of electroacupuncture plus rehabilitation training. Method Seventy-two patients with lumbar intervertebral disc herniation were randomly allocated to a treatment group of 36 cases and a control group of 36 cases. The control group received electroacupuncture at Huatuo jiaji(Ex-B2) points. In addition to electroacupuncture, the treatment group received lumbodorsal muscle stretch training according to the outcome (forward flexion and backward extension) obtained using the Turgunmed testing and evaluating system. Isokinetic muscle strength and the activity were measured and the ODI lumbago score and the VAS pain score were recorded before and after treatment. Result There were statistically significant differences in isokinetic muscle strength and the VAS pain score (P0.05) between the treatment and control groups. Conclusion Electroacupuncture plus rehabilitation training is more effective than electroacupuncture alone in improving isokinetic muscle strength and the VAS pain score in patients with lumbar intervertebral disc herniation.

Role of scaffolding protein JLP on the progression of renal interstitial fibrosis in mice model of unilateral ureteral obstruction and its underlying mechanism / 中华肾脏病杂志

Objective To observe the effect of JLP deficiency on the progression of renal interstitial fibrosis in mice model of unilateral ureteral obstruction (UUO),and to investigate the role and underlying mechanism of JLP in the development of renal fibrosis in obstructive nephropathy.Methods jlp Wild type (jlp+/+) and jlp deficient (jlp-/-) mice were divided into four groups:jlp+/+-and jlp-/--sham-operated groups(jlp-/--sham group and jlp+/+-sham group),jlp+/+-and jlp-/--unilateral ureteral obstruction (UUO)-operated groups (jlp/--UUO group and jlp+/+-UUO group).Mice were sacrificed at 7 days and 14 days after the operation respectively to evaluate the fibrosis by Masson staining.The expression of JLP in jlp +/+ renal tissue was assayed by immunohistochemistry staining,immunofluorescence and Western blotting.Immunohistochemical staining was used to detect the expression of α-smooth muscle actin (α-SMA),collagen Ⅰ (COL-Ⅰ),collagen Ⅲ (COL-Ⅲ) and transforming growth factor-β1 (TGF-β1) in sham and UUO groups.Besides,the α-SMA,COL-Ⅰ,COL-Ⅲ,TGF-β1,p-Smad2 and p-Smad3 protein levels were also analyzed by Western blotting in four groups.Results The expression of JLP was mainly demonstrated in the renal tubules of mice.A large amount of collagen deposition was observed in the renal interstitial area in jlp-/--UUO group compared to jlp+/+-UUO group.Similarly,the expression of α-SMA,COL-Ⅰ,COL-Ⅲ and TGF-β1 was significantly increased in the kidney cortices in jlp-/--UUO-operated groups.Meanwhile,Western blotting showed that the expression of α-SMA,COL-Ⅰ,COL-Ⅲ,and TGF-β1 protein was obviously higher in jlp-/--UUO group.Moreover,the expression of p-Smad2 and p-Smad3 protein was markedly higher in jlp-/--UUO group.Conclusion Scaffolding protein JLP is critical in preventing renal fibrosis through the inhibition of TGF-β1 expression and myo-fibroblast production.

Efficacy of Shenshi Qianghuo Dihuang Decoction in rheumatoid arthritis: a randomized controlled trial / 中西医结合学报

In China, patients with rheumatoid arthritis (RA) are often treated with traditional Chinese herbal medicine. There are certain advantages of traditional Chinese medicine therapy in treatment of RA.

Effects of Yanghe Decoction on vascular endothelial growth factor in cartilage cells of osteoarthritis rabbits / 中西医结合学报

OBJECTIVE: To observe the effects of Yanghe Decoction on vascular endothelial growth factor (VEGF) in cartilage cells of osteoarthritis rabbits. METHODS: Fifteen New Zealand white rabbits were randomly divided into normal group, untreated group and Yanghe Decoction-treated group. The rabbit model of osteoarthritis was established according to Hulth's method. The rabbits were sacrificed at the 8th week after administration of Yanghe Decoction for 14 days, and then rabbit tibia articular cartilage was removed. Sections of the cartilage were stained with Safranin O for histological examination. The cartilage histological characteristics were observed according to the method of Mankin. Immunohistochemical staining was performed to investigate the expression of VEGF. Articular cartilages were observed with microscopy and image analysis method was used to measure the expression intensity of VEGF. RESULTS: There were significant differences in Mankin score between normal group and untreated group (P<0.01), and between untreated group and Yanghe Decoction-treated group (P<0.01). Immunohistochemical staining indicated that the expression intensity of VEGF in untreated group was significantly increased as compared with that in normal group (P<0.01), and also obviously higher than Yanghe Decotion-treated group. CONCLUSION: VEGF plays an important role during early stage of OA. Yanghe Decoction can protect the articular cartilage through suppressing the VEGF expression in chondrocytes and then suppress angiogenesis.

Repair of skin defect in penis with scrotal skin flap / 中华医学美学美容杂志

Objective To explore a simple and convenient method for the treatment of penis skin defect.Methods Since Jan.1995,the scrotal skin flaps have been used to repair the defects of the penis skin in 8 patients with penis skin infection.6 of these cases lost their whole penis skin,and 2 of them lost 2/3 of their penis skin.The granulation tissues in the defective area of penis skin were cut away,and then the normal aponeurosis of corpus cavernosum or spongiosum exposed.The scrotal skin flaps were designed and shaped.The flaps were transferred to cover the defects,and the incisions were closed.Results All the flaps in 8 cases survived completely.The follow-up period was 3 to 12 months after operation.All cases obtained satisfactory results to the contour and function of the penises.Conclusion This procedure is a much better method for repairing skin defects in the penis.It can be easily operated and completed in one-stage.The postoperative results are satisfactory.