Objective To investigate the differences and the immunocompatibility of wild-type (WT), four-gene modified (TKO/hCD55) and six-gene modified (TKO/hCD55/hCD46/hTBM) pig erythrocytes with human serum. Methods The blood samples were collected from 20 volunteers with different blood groups. WT, TKO/hCD55, TKO/hCD55/hCD46/hTBM pig erythrocytes, ABO-compatible (ABO-C) and ABO-incompatible (ABO-I) human erythrocytes were exposed to human serum of different blood groups, respectively. The blood agglutination and antigen-antibody binding levels (IgG, IgM) and complement-dependent cytotoxicity were detected. The immunocompatibility of two types of genetically modified pig erythrocytes with human serum was evaluated. Results No significant blood agglutination was observed in the ABO-C group. The blood agglutination levels in the WT and ABO-I groups were higher than those in the TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups (all P<0.001). The level of erythrocyte lysis in the WT group was higher than those in the ABO-C, TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups. The level of erythrocyte lysis in the ABO-I group was higher than those in the TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups (both P<0.01). The pig erythrocyte binding level with IgM and IgG in the TKO/hCD55 group was lower than those in the WT and ABO-I groups. The pig erythrocyte binding level with IgG and IgM in the TKO/hCD55/hCD46/hTBM group was lower than that in the WT group and pig erythrocyte binding level with IgG was lower than that in the ABO-I group (all P<0.05). Conclusions The immunocompatibility of genetically modified pig erythrocytes is better than that of wild-type pigs and close to that of ABO-C pigs. Humanized pig erythrocytes may be considered as a blood source when blood sources are extremely scarce.

Objective To investigate the establishment of a six-gene-edited pig-to-non-human primate kidney xenotransplantation model. Methods The kidney of humanized genetically-edited pig (GTKO/β4GalNT2KO/CMAHKO/hCD55/hCD46/hTBM) was transplanted into a cynomolgus monkey. The survival of the recipient and kidney condition after blood perfusion were observed. The parenchymal echo, blood flow changes, and size of the kidney were monitored on a regular basis. Routine blood test, kidney function test and electrolyte assessment were carried out. Dynamic changes of urine, feces and body mass were monitored. At the end of life, the transplant kidney, heart, liver, spleen, lung, and cecum were collected for pathological examination. Results The recipient died at postoperative 7 d. After blood flow was restored, the kidney was properly perfused, the organ was soft and the color was normal. At the end of the recipient's life, a slight amount of purulent secretion was attached to the ventral side of the kidney, with evident congestion and swelling, showing the appearance of "red kidney". Postoperatively, the echo of renal parenchyma was increased, blood flow was decreased, the cortex was gradually thickened, and a slight amount of effusion surrounded the kidney and abdominal cavity over time. In the recipient, the amount of peripheral red blood cells, hemoglobin, albumin, and platelets was progressively decreased, and serum creatinine level was increased to 308 μmol/L at postoperative 7 d, whereas the K⁺ concentration did not significantly change. Light yellow urine was discharged immediately after surgery, diet and drinking water were resumed within postoperative 3 h, and light yellow and normal-shape stool was discharged. The reddish urine was gradually restored to normal color within postoperative 1 d, which were consistent with the results of the routine urine test. A large amount of brown bloody stool was discharged twice in the morning of 2 d after surgery. Omeprazole was given for acid suppression, and the stool returned to normal at postoperative 4 d. The β₂-microglobulin level was increased to 0.75 mg/L at postoperative 7 d. The body mass was increased by 1.7 kg. Autopsy pathological examination showed interstitial edema and bleeding of the transplant kidney, a large amount of infiltration of lymphocytes and macrophages, infiltration of lymphocytes in the arteriole wall and arterial cavity, accompanied by arteritis changes, lymphocyte infiltration in the cecal stroma and congestion in the spleen tissues. No significant abnormal changes were observed in other organs. Conclusions The humanized genetically-edited pig-to-non-human primate kidney xenotransplantation model is successfully established, and postoperative survival of the recipient is 1 week.

Cardiovascular diseases (CVDs) and metabolic disorders are major components of noncommunicable diseases, causing an enormous health and economic burden worldwide. There are common risk factors and developmental mechanisms among them, indicating the far-reaching significance in exploring the corresponding therapeutic targets. MST1/2 kinases are well-established proapoptotic effectors that also bidirectionally regulate autophagic activity. Recent studies have demonstrated that MST1/2 influence the outcome of cardiovascular and metabolic diseases by regulating immune inflammation. In addition, drug development against them is in full swing. In this review, we mainly describe the roles and mechanisms of MST1/2 in apoptosis and autophagy in cardiovascular and metabolic events as well as emphasis on the existing evidence for their involvement in immune inflammation. Moreover, we summarize the latest progress of pharmacotherapy targeting MST1/2 and propose a new mode of drug combination therapy, which may be beneficial to seek more effective strategies to prevent and treat CVDs and metabolic disorders.

Objective:To evaluate the clinical effect of Guanxin Tongmai plaster combined with conventional western medicine in the treatment of phlegm and blood stasis syndrome of coronary heart disease and angina pectoris.Methods:A total of 60 patients in the Department of Cardiology of the Affiliated Hospital of Liaoning University of Traditional Chinese Medicine (TCM) from February to August 2020 who met the inclusion criteria were randomly divided into two groups with 30 in each group. Both groups were treated with conventional western medicine. On this basis, Guanxin Tongmai plaster was applied at the acupoints in the treatment group and placebo plaster was applied in the control group. TCM syndrome score was performed before and after treatment, angina score was evaluated from three aspects of angina attack frequency, duration and pain degree, and blood lipid TG, TC, LDL-C and HDL-C were detected by enzyme quantitative method. The blood homocysteine (Hcy) was detected by enzyme circulation method, the ECG and the nitroglycerin reduction rate were recorded, and the safety index was detected according to the ECG changes.Results:In the treatment period, 2 patients in the treatment group fell off, 3 in the control group. A total of 28 patients in the treatment group and 27 in the control group were analyzed. The total effective rate of ECG efficacy in the treatment group was 67.9% (19/28) and the control group was 48.1% (13/27). There was significant difference between the two groups ( χ2=4.46, P=0.040). After treatment, the TCM syndrome score and angina score in the treatment group were significantly lower than those in the control group ( t values were 9.12 and 4.45, P values were 0.004 and 0.042, respectively). The reduction rate of nitroglycerin in the treatment group was 82.1% (23/28) and 55.6% (15/27) in the control group. There was significant difference between the two groups ( χ 2=4.72, P=0.030). After treatment, the plasma TG, TC, LDL-C in the treatment group were significantly lower than those in the control group ( t values were 4.17, 6.57 and 6.52, P<0.05 or P<0.01), the level of HDL-C was significantly higher than that of the control group ( t=7.07, P=0.010), and the level of plasma Hcy was significantly lower than that in the control group ( t=6.70, P=0.012). There was no significant difference in liver, kidney and coagulation function between the two groups. Conclusion:Guanxin Tongmai plaster combined with conventional western medicine can improve the clinical symptoms of patients with coronary heart disease and angina pectoris, reduce the level of blood lipid and Hcy, and improve the clinical curative effect.

Objective:To explore the epidemiological characteristics of patients with electrical burn at different voltages complicated by cerebral trauma, so as to provide a basis for improving the treatment level of such injury.Methods:A retrospective cohort study was conducted to analyze the clinical data of 480 patients with electrical burn complicated by cerebral trauma treated in Qingdao Municipal Hospital affiliated to Qingdao University from January 2001 to December 2019. According to the voltage intensity, the patients were divided into low voltage group (injury voltage<1 kV, n=295) and high voltage group (injury voltage≥1 kV, n=185). Gender, age, status of burn and other general data of all patients were collected. The clinical manifestations, consciousness [Glasgow coma scale (GCS)], imaging findings, treatment, prognosis [Glasgow outcome scale (GOS)] and complications were compared between the two groups. Results:(1) Gender and age: the male to female ratio was 5.4∶1.0; the peak age of onset occurred at 18-60 years, accounting for 302 patients (62.9%); the status of burn: the burn area ranged from 1%-78% [(20.0±4.0)%] total body surface area (TBSA), with the current outlet located at the head in 321 patients. (2) Clinical manifestations: consciousness disorders accounted for the highest proportion, with 295 patients (100%) in low voltage group and 185 patients (100%) in high voltage group, followed by headache which occurred in 178 patients (60.3%) in low voltage group and 115 patients (62.2%) in high voltage group (all P>0.05). (3) Consciousness: 37 patients presented coma, with 17 patients (5.8%) in low voltage group and 20 patients (10.8%) in high voltage group ( P<0.05). (4) Imaging findings: CT and MRI examination found cerebral edema, skull fracture, intracranial hematoma, cerebral ischemia, subarachnoid hemorrhage, and other positive lesions. In patients with head wounds (current inlet and outlet located in the head), the incidence of cerebral trauma was 44.0% in low voltage group and 86.8% in high voltage group ( P<0.05). In patients with no head wound (the current outlet was not located in the head), the incidence of cerebral trauma was 5.3% in low voltage group and 6.3% in high voltage group ( P>0.05). In contrast with the patients without current outlet locating in the head, there were three more types of cerebral trauma in patients with current outlet locating in the head, including skull fracture, intracranial hematoma and subarachnoid hemorrhage. (5) Treatment, prognosis and complications: 478 patients (99.6%) received non-surgical treatment and 2 patients (0.4%) received surgical treatment. There was 1 death (0.2%) and 479 successfully treated patients (99.8%). The prognosis was good in 280 patients (94.9%), moderately disabled in 13(4.4%) and severely disabled in 2(0.7%) in low voltage group; while the prognosis was good in 143 patients (77.3%), moderately disabled in 30(16.2%), severely disabled in 11(5.9%) and death in 1(0.5%) in high voltage group (all P<0.01). After discharge, the incidence of numbness, paresthesia and anxiety was significantly higher in low voltage group than that in high voltage group (all P<0.01). Conclusions:Male patients with electrical burn complicated by cerebral trauma are more than female patients, with the young and middle-aged population being at high risk. Disturbance of consciousness and headache are the main clinical manifestations. The incidence of high voltage coma is relatively higher. Compared with low voltage-induced electrical burn, the patients with high voltage-induced electrical burn complicated by cerebral trauma (current inlet and outlet located at the head) sustain more severe and extensive injury. Early and active CT or MRI examination is conducive to definite diagnosis. Non-surgical treatment is the main treatment. Compared with high voltage-induced electrical burn, the patients with low voltage-induced electrical burn complicated by cerebral trauma have significantly better prognosis, but are more likely to develop complications of numbness, paresthesia and anxiety.

Objective:To observe the expressions of complement 3 (C3) and endothelial cell injury-associated proteins before and after cathepsin L (CTSL) blockade in renal injury of trichloroethylene (TCE) -sensitized mice.Methods:In June 2018, 41 SPF female BALB/c mice were divided respectively into blank control group ( n=5) , vehicle control group ( n=5) , TCE group ( n=15) and TCE+CTSLi group ( n=16) to establish trichloroethylene-sensitized mice model by pretreating the mice with intraperitoneal injection of CTSL inhibitor (CTSLi) and using TCE for the first and last challenge. According to the skin sensitization score, the mice were divided into positive group and negative group. 72 hours after the last challenge, the renal function indexes of the mice were detected, the pathological changes of mice kidneys were observed, and the glomerular C3 and endothelial cell damage-related proteins [vascular cell adhesion molecule 1 (VCAM-1) , tight junction protein 5 (Claudin-5) and Syndecan-1] expression levels were detected. Results:The sensitization rates of mice in TCE group and TCE+CTSLi group were 53.3% (8/15) and 50.0% (8/16) , respectively, and there was no significant difference between the two groups ( P>0.05) . Compared with vehicle control group and the corresponding TCE negative group, the serum creatinine (CRE) and blood urea nitrogen (BUN) levels of mice in the TCE positive group was increased, while the TCE positive group were higher than the TCE+CTSLi positive group ( P<0.05) . Pathological examination showed obvious vacuolar degeneration and cellular edema in the mice kidney of the TCE positive group. In the TCE+CTSLi positive group, the above pathological damage was significantly improved. Immunohistochemical results showed that the expression of glomerular C3 fragment and VCAM-1 in TCE positive group were significantly higher than that of the vehicle control and TCE negative group ( P<0.05) , while TCE+CTSLi positive group was significantly lower than that of TCE positive group ( P<0.05) . Western blot test results showed that the relative expression levels of Claudin-5 and Syndecan-1 protein in the mice glomeruli of TCE positive group were significantly lower than those in the vehicle control group and TCE negative group ( P<0.05) . Compared with the TCE positive group, the Claudin-5 protein was increased in the kidney of the TCE+CTSLi positive group, but the difference was not statistically significant ( P>0.05) , while the Syndecan-1 protein was significantly increased in the TCE+CTSLi positive group ( P<0.05) . Conclusion:CTSL may mediate the glomerular structural damage by cutting complement C3, activating the complement system, damaging endothelial cell structural protein Syndecan-1 and overexpressing adhesion molecule VCAM-1 in TCE-sensitized mice. Inhibiting the expression of CTSL may be an effective way to protect the glomerular integrity of structure and function in pharmacology.

Objective:To observe the expressions of complement 3 (C3) and endothelial cell injury-associated proteins before and after cathepsin L (CTSL) blockade in renal injury of trichloroethylene (TCE) -sensitized mice.Methods:In June 2018, 41 SPF female BALB/c mice were divided respectively into blank control group ( n=5) , vehicle control group ( n=5) , TCE group ( n=15) and TCE+CTSLi group ( n=16) to establish trichloroethylene-sensitized mice model by pretreating the mice with intraperitoneal injection of CTSL inhibitor (CTSLi) and using TCE for the first and last challenge. According to the skin sensitization score, the mice were divided into positive group and negative group. 72 hours after the last challenge, the renal function indexes of the mice were detected, the pathological changes of mice kidneys were observed, and the glomerular C3 and endothelial cell damage-related proteins [vascular cell adhesion molecule 1 (VCAM-1) , tight junction protein 5 (Claudin-5) and Syndecan-1] expression levels were detected. Results:The sensitization rates of mice in TCE group and TCE+CTSLi group were 53.3% (8/15) and 50.0% (8/16) , respectively, and there was no significant difference between the two groups ( P>0.05) . Compared with vehicle control group and the corresponding TCE negative group, the serum creatinine (CRE) and blood urea nitrogen (BUN) levels of mice in the TCE positive group was increased, while the TCE positive group were higher than the TCE+CTSLi positive group ( P<0.05) . Pathological examination showed obvious vacuolar degeneration and cellular edema in the mice kidney of the TCE positive group. In the TCE+CTSLi positive group, the above pathological damage was significantly improved. Immunohistochemical results showed that the expression of glomerular C3 fragment and VCAM-1 in TCE positive group were significantly higher than that of the vehicle control and TCE negative group ( P<0.05) , while TCE+CTSLi positive group was significantly lower than that of TCE positive group ( P<0.05) . Western blot test results showed that the relative expression levels of Claudin-5 and Syndecan-1 protein in the mice glomeruli of TCE positive group were significantly lower than those in the vehicle control group and TCE negative group ( P<0.05) . Compared with the TCE positive group, the Claudin-5 protein was increased in the kidney of the TCE+CTSLi positive group, but the difference was not statistically significant ( P>0.05) , while the Syndecan-1 protein was significantly increased in the TCE+CTSLi positive group ( P<0.05) . Conclusion:CTSL may mediate the glomerular structural damage by cutting complement C3, activating the complement system, damaging endothelial cell structural protein Syndecan-1 and overexpressing adhesion molecule VCAM-1 in TCE-sensitized mice. Inhibiting the expression of CTSL may be an effective way to protect the glomerular integrity of structure and function in pharmacology.

Pace of life and work of people is accelerating nowadays, and hospitals keep improving their services, which gives rise to the decoction and delivery service from the third party vendors for traditional Chinese medicine(TCM).Given the quality control standards for the TCM decoction service as issued by Zhejiang and other regions, the systems of supervision and assessment remain incomplete.Authors of the paper introduced a project improvement team, composed of Chinese medicine pharmacy, Chinese medicine experience specialists, vendors of Chinese medicine decoction and express delivery companies, hence establishing a " three-in-one" supervision system of Internet+TCM decoction and delivery service. This practice can optimize the assessment indexes, strengthen the assessment system of assessment transformation and supervision system for patient medication.It proves that the practice contributes to higher quality and safety of TCM decoction and delivery service, improves the ability and level of TCM services, and ensures the medication safety of patients.

Objective: To explore the long-term efficacy of laparoscopic-assisted anorectoplasty and conventional anorectoplasty in the treatment of children with high and middle anal atresia. Methods: A retrospective cohort study was used. Inclusion criteria: (1) children with high and middle anal atresia; (2) complicated with rectourethral or rectovesical fistula; (3) complete follow-up data. Exclusion criteria: (1) complicated with 21-trisomy; (2) cerebral palsy and other mentaldisabilities; (3) Currarino syndrome; (4) FG syndrome. Clinical data of 88 patients with middle and high anal atresia, who complicated with rectourethral fistula or rectovesical fistula, and underwent anoplasty at Department of Pediatric Surgery, the First Affiliated Hospital of Zhengzhou University from January 2009 to June 2014 were enrolled in the study and analyzed. There were 24 cases with middle atresia and 64 cases with high atresia. All the cases were divided into 2 groups based on the operative method: laparoscopic group (laparoscopic-assisted anorectoplasty, 49 cases), pena group (posterior sagittal anorectoplasty, 39 cases). The demographic features of two groups were comparable. There were no statistically significant differences in gender, age, body mass, classification of anomaly types and sacral ratio (all P>0.05). Student t test and Chi square tests were used to compare the surgical conditions (operative time, postoperative hospital stay and complications), anal function (Kelly score), constipation (Krickenbeck constipation score) and anorectal pressure. Results: Children of both groups all completed operation ssuccessfully. There were no statistically significant differences between laparoscopic group and pena group in the operative time [(120±31) minutes vs. (112±23) minutes, t=1.343, P=0.091] and postoperative hospital stay [(7.1±2.3) days vs. (10.7±3.3) days, t=6.021, P=1.000]. Complications were more common in the pena group [16.3% (8/49) vs. 35.9% (14/39), χ²=4.436, P=0.035]. The main complications in laparoscopic group were anal prolapse (8.2%, 4/49) and anal stenosis (6.2%, 3/49), while in pena group were anal stenosis (12.8%, 5/39) and perioperative perianal skin erosion (10.3%, 4/39). As for the anal function, the degree of feces, defecation control and sphincter contractility, the single scoring differences of Kelly scoring system were not statistically significant between the two groups, but the proportion of good function in the laparoscopic group was higher than that in the pena group [67.3% (8/49) vs. 38.5% (15/39), χ²=7.308, P=0.007]. Constipation occurred in 6 (12.2%) patients in the laparoscopic group, of whom 5 were improved by diet regulation and 1 required laxatives, while 9 (23.1%) patients developed constipation in the pena group, of whom 4 were improved by diet regulation and 5 required long-term laxatives. The difference of constipation ratio was not statistically significant (χ²=1.802, P=0.180). There were no cases of Krickenbeck constipation grade 3. Compared to the pena group, the laparoscopic group had higher anal resting pressure [(33.35±9.69) mmHg vs. (27.68±10.74) mmHg, t=2.599, P=0.011], higher dilating pressure [(9.00±5.61) mmHg vs.(6.51±3.24) mmHg, t=2.462, P=0.016], higher maximal squeeze pressure [(65.80±17.23) mmHg vs. (56.74±18.93) mmHg, t=2.389, P=0.019] and longer maximal contraction time [(21.16±5.02) seconds vs. (18.44±7.24) seconds, t=2.079, P=0.041]. The rectal resting pressure [(5.36±3.00) mmHg vs. (4.61±3.93) mmHg, t=1.015, P=0.312] was not statistically significantly different. Conclusions Compared with posterior sagittal anorectoplasty, laparoscopic-assisted anorectoplasty in the treatment of high and middle anal atresia has better long-term efficacy with less perioperative complications.

Objective To explore the long?term efficacy of laparoscopic?assisted anorectoplasty and conventional anorectoplasty in the treatment of children with high and middle anal atresia. Methods A retrospective cohort study was used. Inclusion criteria: (1) children with high and middle anal atresia; (2) complicated with rectourethral or rectovesical fistula; (3) complete follow?up data. Exclusion criteria: (1) complicated with 21?trisomy; (2) cerebral palsy and other mentaldisabilities; (3) Currarino syndrome; (4) FG syndrome. Clinical data of 88 patients with middle and high anal atresia, who complicated with rectourethral fistula or rectovesical fistula, and underwent anoplasty at Department of Pediatric Surgery, the First Affiliated Hospital of Zhengzhou University from January 2009 to June 2014 were enrolled in the study and analyzed. There were 24 cases with middle atresia and 64 cases with high atresia. All the cases were divided into 2 groups based on the operative method: laparoscopic group (laparoscopic?assisted anorectoplasty, 49 cases), pena group (posterior sagittal anorectoplasty, 39 cases). The demographic features of two groups were comparable. There were no statistically significant differences in gender, age, body mass, classification of anomaly types and sacral ratio (all P>0.05). Student t test and Chi square tests were used to compare the surgical conditions (operative time, postoperative hospital stay and complications), anal function (Kelly score), constipation (Krickenbeck constipation score) and anorectal pressure. Results Children of both groups all completed operation ssuccessfully. There were no statistically significant differences between laparoscopic group and pena group in the operative time [(120±31) minutes vs. (112±23) minutes, t=1.343, P=0.091] and postoperative hospital stay [(7.1±2.3) days vs. (10.7±3.3) days, t=6.021, P=1.000]. Complications were more common in the pena group [16.3% (8/49) vs. 35.9% (14/39), χ2=4.436, P=0.035]. The main complications in laparoscopic group were anal prolapse (8.2%, 4/49) and anal stenosis (6.2%, 3/49), while in pena group were anal stenosis (12.8%, 5/39) and perioperative perianal skin erosion (10.3%, 4/39). As for the anal function, the degree of feces, defecation control and sphincter contractility, the single scoring differences of Kelly scoring system were not statistically significant between the two groups, but the proportion of good function in the laparoscopic group was higher than that in the pena group [67.3% (8/49) vs. 38.5% (15/39), χ2=7.308, P=0.007]. Constipation occurred in 6 (12.2%) patients in the laparoscopic group, of whom 5 were improved by diet regulation and 1 required laxatives, while 9 (23.1%) patients developed constipation in the pena group, of whom 4 were improved by diet regulation and 5 required long?term laxatives. The difference of constipation ratio was not statistically significant (χ2=1.802, P=0.180). There were no cases of Krickenbeck constipation grade 3. Compared to the pena group, the laparoscopic group had higher anal resting pressure [(33.35 ± 9.69) mmHg vs. (27.68 ± 10.74) mmHg, t=2.599, P=0.011], higher dilating pressure [(9.00±5.61) mmHg vs.(6.51±3.24) mmHg, t=2.462, P=0.016], higher maximal squeeze pressure [(65.80 ± 17.23) mmHg vs. (56.74 ± 18.93) mmHg, t=2.389, P=0.019] and longer maximal contraction time [(21.16±5.02) seconds vs. (18.44±7.24) seconds, t=2.079, P=0.041]. The rectal resting pressure [(5.36 ± 3.00) mmHg vs. (4.61 ± 3.93) mmHg, t=1.015, P=0.312] was not statistically significantly different. Conclusions Compared with posterior sagittal anorectoplasty, laparoscopic?assisted anorectoplasty in the treatment of high and middle anal atresia has better long?term efficacy with less perioperative complications.