Objective: To evaluate the effects of green tea extract (GTE) and its main catechin monomers on erythrocyte lesion (such as hemolysis, decreased energy metabolism and oxidative stress) during in vitro storage, and to explore its potential as a novel additive for erythrocyte preservation solutions. Methods: The composition of GTE was analyzed by high-performance liquid chromatography (HPLC). Using an in vitro simulated storage model, erythrocytes were stored in CPDA-1 preservation solution supplemented with GTE and the three most abundant catechin monomers (EGCG, ECG, EGC) for 60 days. Hemolysis rate and ATP content were dynamically monitored during storage. Flow cytometry was used to analyze phosphatidylserine (PS) exposure. Meanwhile, the protective effects of each component were verified in an acute oxidative stress model, and erythrocyte membrane stability was assessed by osmotic fragility test. Results: After 60 days of storage at 4℃, the hemolysis rate at the end of storage in the GTE group was <0.8%, which was superior to that in the control group and the single catechin-supplemented groups. Erythrocyte osmotic fragility assay showed that GTE could enhance the stability of erythrocyte membranes. In the acute oxidative stress experiment, the protective rate of GTE against erythrocyte injury exceeded 99%, which was better than that of the single catechin groups. At the initial stage of storage, ATP content decreased in all catechin-treated groups, but PS exposure was not significantly increased. Conclusion: The addition of GTE can effectively alleviate storage lesions of erythrocytes, with efficacy superior to that of single catechins. GTE enhances the antioxidant capacity and membrane stability of stored erythrocytes. Our results provide new experimental evidence for the development of GTE-based erythrocyte preservation additives.

Objective To discuss the factors associated with residual shunt occurring after closure surgery of patent foramen ovale(PFO,and to analyze the relationship between the residual shunt and the relief degree of migraine.Methods A total of 210 patients with PFO,who received closure surgery of PFO due to the occurrence of premonitory migraine at the Jining Municipal First People's Hospital of China from January 2020 to December 2022,were collected.The clinical data,visual analogue scale(VAS)score,and headache impact test-6 questionnaire(HIT-6)score of patients having postoperative residual shunt and patients having no postoperative residual shunt were analyzed.Results A total of 172 patients had no postoperative residual shunt(no-shunt group),and 38 patients had postoperative residual shunt(shunt group).The preoperative PFO width in the shunt group was(1.72±0.34)mm,which was obviously larger than(1.42±0.31)mm in the no-shunt group(P＜0.05).The proportion of patients with right-to-left shunt grades Ⅰ-Ⅱ in resting state in the shunt group was 94.74％,which was higher than 65.21％in the no-shunt group(P＜0.05).No statistically significant differences in gender,age,body mass index(BMI),diabetes,hypertension,hyperlipemia,operation time and occluder type existed between the two groups(P＞0.05).Logistic regression analysis showed that preoperative PFO width and the grade of right-to-left shunt in resting state were the influencing factors for the occurrence of residual shunt(P＜0.05).Based on this model,a column chart model for predicting residual shunt was established,and its predicted area under the ROC curve was 0.882(95％CI:0.731-0.944),the sensitivity was 82.50％,and specificity was 81.00％.The postoperative 12-month VAS score and HIT-6 score in the shunt group were(1.87±0.29)points and(56.02±9.08)points respectively,which were remarkably higher than those in the no-shunt group(P＜0.05).The improvement rates of VAS score and HIT-6 score in the shunt group were(72.59±9.17)％and(22.43±8.11)％respectively,which was lower than those in the no-shunt group(P＜0.05).The postoperative 12-month VAS score and HIT-6 score in the patients having severe residual shunt were(2.21±0.41)points and(61.38±8.45)points respectively,which were higher than those in the patients having mild residual shunt(P＜0.05).The residual shunt volume was negatively correlated with the improvement rates of VAS score and HIT-6 score(r=-0.546 and r=-0.738 respectively,both P＜0.05).Conclusion Residual shunt volume is negatively associated with the relief degree of migraine in patients after receiving closure surgery of PFO due to the occurrence of premonitory migraine.

Background: Radical hysterectomy (RH) is commonly used to treat early-stage cervical cancer in women of childbearing age and sexual dysfunction due to postoperative vaginal shortening is a major concern. The impact of intraoperative vaginoplasty on prognosis and quality of sexual life in patients with early-stage cervical cancer remains controversial and lacks high-level evidence. However, there are few reports on vaginoplasty after RH to lengthen vagina in patients. This prospective, multi-center, randomized controlled trial aims to explore the impact of peritoneal vaginoplasty with or without ovarian transposition after laparoscopic RH on sexual dysfunction in patients with early-stage cervical cancer. Methods Eligible patients will be randomly assigned (1:1) to receive peritoneal vaginoplasty or not. The primary evaluation indicators are female sexual function index (FSFI) and male sexual satisfaction scale. The secondary evaluation indicators include EORTC QLQ-CX24, 2-year overall survival (OS), 5-year OS, 2-year progression-free survival (PFS), 5-year PFS and surgery-related complications. The trial will enroll 368 patients from 6 hospitals in China over a 3-year period and follow up for 5 years.Trial RegistrationChinese Clinical Trial Registry Identifier: ChiCTR2000040610SynopsisIt's a prospective, randomized controlled trial to explore the effect of peritoneal vaginoplasty with or without ovarian transposition following laparoscopic radical hysterectomy on sexual dysfunction in patients with cervical cancer. The trial will enroll 368 patients from 6 hospitals in China over a 3-year period and follow up for 5 years.

Objective To retrospectively analyze the blood use of transfusion-dependent thalassemia(TDT)patients in 9 designated transfusion medical institutions from 2018 to 2023 in Nanning,and to evaluate the effect of"three designated"blood transfusion mode(hereby means TDT patients undergoing blood transfusion in designated transfusion medical institu-tions regularly)and"collection-based-supply"blood management mode on blood security of TDT patients.Methods The"three designated"blood transfusion mode was implemented to ensure that TDT patients registered in the local household registration(referred to as the"register")obtain the rights and interests of outpatient transfusion and blood security of des-ignated medical institutions.The"collection-based-supply"blood management mode was implemented to assess the blood needs of"register"TDT patients and meet their needs to the maximum extent according to the blood inventory(collection).Results From 2018 to 2023,the total blood supply of"register"TDT patients was 10.37%of the total red blood supply of all medical institutions(138 509.5 U/1 335 788.0 U),with the highest proportion of type O blood as 46.34%(64 181.0 U/138 509.5 U)and the lowest proportion of type AB blood as 3.85%(5 331.0 U/138 509.5 U).In 2018,9 transfusion medical institutions were designated for TDT patients.There were a total of 766 TDT patients in the register,with the per ca-pita annual blood transfusion volume increased from20.28 U(15 531.0 U/766 patients)in2018 to36.01 U(27 586.0 U/766 patients)in 2023,maintaining a positive growth every year(30.26%,4.94%,11.71%,8.61%,4.94%and 7.10%).Conclusion The"three designated"blood transfusion mode and the"collection-based-supply"blood management mode can effectively guarantee the blood supply of TDT patients.

Background: Radical hysterectomy (RH) is commonly used to treat early-stage cervical cancer in women of childbearing age and sexual dysfunction due to postoperative vaginal shortening is a major concern. The impact of intraoperative vaginoplasty on prognosis and quality of sexual life in patients with early-stage cervical cancer remains controversial and lacks high-level evidence. However, there are few reports on vaginoplasty after RH to lengthen vagina in patients. This prospective, multi-center, randomized controlled trial aims to explore the impact of peritoneal vaginoplasty with or without ovarian transposition after laparoscopic RH on sexual dysfunction in patients with early-stage cervical cancer. Methods Eligible patients will be randomly assigned (1:1) to receive peritoneal vaginoplasty or not. The primary evaluation indicators are female sexual function index (FSFI) and male sexual satisfaction scale. The secondary evaluation indicators include EORTC QLQ-CX24, 2-year overall survival (OS), 5-year OS, 2-year progression-free survival (PFS), 5-year PFS and surgery-related complications. The trial will enroll 368 patients from 6 hospitals in China over a 3-year period and follow up for 5 years.Trial RegistrationChinese Clinical Trial Registry Identifier: ChiCTR2000040610SynopsisIt's a prospective, randomized controlled trial to explore the effect of peritoneal vaginoplasty with or without ovarian transposition following laparoscopic radical hysterectomy on sexual dysfunction in patients with cervical cancer. The trial will enroll 368 patients from 6 hospitals in China over a 3-year period and follow up for 5 years.

Background: Radical hysterectomy (RH) is commonly used to treat early-stage cervical cancer in women of childbearing age and sexual dysfunction due to postoperative vaginal shortening is a major concern. The impact of intraoperative vaginoplasty on prognosis and quality of sexual life in patients with early-stage cervical cancer remains controversial and lacks high-level evidence. However, there are few reports on vaginoplasty after RH to lengthen vagina in patients. This prospective, multi-center, randomized controlled trial aims to explore the impact of peritoneal vaginoplasty with or without ovarian transposition after laparoscopic RH on sexual dysfunction in patients with early-stage cervical cancer. Methods Eligible patients will be randomly assigned (1:1) to receive peritoneal vaginoplasty or not. The primary evaluation indicators are female sexual function index (FSFI) and male sexual satisfaction scale. The secondary evaluation indicators include EORTC QLQ-CX24, 2-year overall survival (OS), 5-year OS, 2-year progression-free survival (PFS), 5-year PFS and surgery-related complications. The trial will enroll 368 patients from 6 hospitals in China over a 3-year period and follow up for 5 years.Trial RegistrationChinese Clinical Trial Registry Identifier: ChiCTR2000040610SynopsisIt's a prospective, randomized controlled trial to explore the effect of peritoneal vaginoplasty with or without ovarian transposition following laparoscopic radical hysterectomy on sexual dysfunction in patients with cervical cancer. The trial will enroll 368 patients from 6 hospitals in China over a 3-year period and follow up for 5 years.

ObjectiveTo investigate the protective effect of modified Huangqi Guizhi Wuwutang （MHGW） on endoplasmic reticulum stress in the sciatic nerve of diabetes rats based on the pathways of inositol-requiring enzyme 1α （IRE1α） and CCAAT/enhancer-binding protein homologous protein （CHOP）. MethodSixty rats were fed on a high-sugar and high-fat diet for six weeks， followed by intraperitoneal injection of streptozotocin at a dose of 35 mg·kg^-1. Random blood glucose levels were measured three days later and rats with a sustained blood glucose level ≥ 16.7 mmol·L^-1 were included in study （n=48）. The rats were randomly divided into a model group， an α-lipoic acid group （0.026 8 g·kg^-1·d^-1）， a high-dose MHGW group （2.5 g·kg^-1·d^-1）， and a low-dose MHGW group （1.25 g·kg^-1·d^-1）. Another 10 rats were assigned to the normal group. The intervention lasted for 16 weeks. After 16 weeks， the sciatic nerve structure of the rats in each group was observed under light microscopy using Luxol fast blue （LFB） staining. Transmission electron microscopy was used to observe the ultrastructure of the sciatic nerve. Chemiluminescence method was employed to measure the serum reactive oxygen species （ROS） levels. Western blot and real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） were used to evaluate the expression of p-IRE1α protein， IRE1α mRNA， CHOP protein， and CHOP mRNA in the sciatic nerve of the rats. ResultCompared with the normal group， the model group showed elevated serum ROS levels （P<0.01）. In contrast， the serum ROS levels were significantly reduced in the treatment groups compared with those in the model group （P<0.01）. The sciatic nerve of the model group showed pathological changes compared with that in the normal group， while the treatment groups exhibited improvement in sciatic nerve pathology compared with the model group. The protein expression of p-IRE1α and CHOP in the sciatic nerve significantly increased in the model group as compared with that in the normal group （P<0.01）. However， the treatment groups showed a significant decrease in the protein expression of p-IRE1α and CHOP in the sciatic nerve compared with the model group （P<0.05， P<0.01）. Furthermore， compared with the normal group， the model group showed upregulated mRNA expression of IRE1α and CHOP in the sciatic nerve （P<0.01）， while the treatment groups exhibited a significant decrease in the mRNA expression of IRE1α and CHOP compared with the model group （P<0.01）. ConclusionMHGW can alleviate endoplasmic reticulum stress-induced cell apoptosis and improve the structure and function of the sciatic nerve in diabetes rats by inhibiting the expression of IRE1α/CHOP pathway-related proteins and mRNA， thereby preventing and treating peripheral neuropathy in diabetes.

ObjectiveTo investigate the effect of modified Huangqi Guizhi Wuwutang （MHGW） on the protein and mRNA expression of B-cell lymphoma-2-associated X protein （Bax） and cysteinyl aspartate specific proteinase-12 （Caspase-12） related to the apoptosis of sciatic nerve cells in diabetes rats to explore the mechanism of MHGW in the treatment of peripheral neuropathy in diabetes. MethodAnimal experiments were conducted. A diabetes model was induced in sixty male sprague-dawley （SD） rats by feeding on a high-sugar and high-fat diet combined with streptozotocin （STZ） intraperitoneal injection. Rats with random blood glucose levels ≥ 16.7 mmol·L^-1 for three consecutive days were considered to have successfully developed diabetes. Forty-eight rats that successfully developed diabetes were randomly divided into a model group， an α-lipoic acid group （0.026 8 g·kg^-1·d^-1）， a high-dose MHGW group （2.5 g·kg^-1·d^-1）， and a low-dose MHGW group （1.25 g·kg^-1·d^-1）， with 12 rats in each group. Another 10 rats were assigned to the normal group. Body weight and random blood glucose levels of the rats were monitored. At the end of a 16-week intervention period， the sciatic nerve conduction velocity of the rats was measured using the Key point electromyography collection system. The protein and mRNA expression of Bax and Caspase-12 in the sciatic nerve cells was detected by Western blot analysis and real-time quantitative polymerase chain reaction （Real-time PCR）， respectively. ResultCompared with the normal group， the model group showed a significant decrease in body weight （P<0.01） and a significant increase in random blood glucose levels （P<0.01）. After a 16-week intervention， compared with the model group， the high-dose MHGW group exhibited a significant increase in body weight （P<0.05）， while there were no statistically significant differences in body weight changes among the other treatment groups. Random blood glucose levels significantly decreased in all treatment groups （P<0.01）. After 16 weeks of intervention， compared with the normal group， the model group had significantly reduced motor and sensory nerve conduction velocities （P<0.01）. Compared with the model group， all treatment groups showed significant increases in motor and sensory nerve conduction velocities （P<0.05， P<0.01）. The expression of Bax and Caspase-12 proteins in the sciatic nerve cells was significantly elevated in the model group compared with that in the normal group （P<0.01）. In contrast， all treatment groups showed significant reductions in the expression of Bax and Caspase-12 proteins in the sciatic nerve cells as compared with that in the model group （P<0.01）. The expression of Bax and Caspase-12 mRNA in the sciatic nerve cells significantly increased in the model group compared with that in the normal group （P<0.01）. Compared with the model group， the α-lipoic acid group and the high-dose MHGW group showed significant reductions in the expression of Bax mRNA in the sciatic nerve cells （P<0.05， P<0.01）， while the low-dose MHGW group showed a decreasing trend in the expression of Bax mRNA. The expression of Caspase-12 mRNA in the sciatic nerve cells significantly decreased in all treatment groups （P<0.01）. ConclusionMHGW may improve and repair sciatic nerve damage in diabetes rats by inhibiting sciatic nerve cell apoptosis.

DNA polymerases are widely used in PCR and play important roles in life science research and related fields. Development of high-performance DNA polymerases is of great commercial interest as the current commercial DNA polymerases could not fully satisfy the requirements of scientific research. In this study, we cloned and expressed a family B DNA polymerase (NCBI accession number TEU_RS04875) from Thermococcus eurythermalis A501, characterized its enzymatic property and evaluated its application in PCR. The recombinant Teu-PolB was expressed in E. coli and purified with affinity chromatography and ion-exchange chromatography. The enzymatic properties of Teu-PolB were characterized using fluorescence-labeled oligonucleotides as substrates. The application potential of Teu-PolB in PCR was evaluated using the phage λ genomic DNA as a template. Teu-PolB has DNA polymerase and 3'→5' exonuclease activities, and is highly thermostable with a half-life of 2 h at 98 ℃. The most suitable PCR buffer is consisted of 50 mmol/L Tris-HCl pH 8.0, 2.5 mmol/L MgCl₂, 60 mmol/L KCl, 10 mmol/L (NH₄)₂SO₄, 0.015% Triton X-100 and 0.01% BSA, and the optimal extension temperature is 68 ℃. Under the optimized conditions, a 4 kb target fragment was successfully amplified with an extension rate of 2 kb/min. The yield of the Teu-PolB amplified-DNA was lower than that of Taq DNA polymerase, but its extension rate and fidelity was higher than that of Taq and Pfu DNA polymerases. The biochemical properties of Teu-PolB demonstrate that this enzyme can be used in PCR amplification with high thermostability, good salt tolerance, high extension rate and high fidelity.
DNA-Directed DNA Polymerase/genetics* ; Escherichia coli/genetics* ; Polymerase Chain Reaction/methods* ; Temperature ; Thermococcus/genetics*

Objective To summarize the factors and the corresponding treatments for patients with coronary heart disease (CHD) complicated by thrombocytopenia, and provide medical advises for clinical treatment. Methods Literatures and case reports were analyzed and summarized. Results Thrombocytopenia in patient with CHD could be mainly divided into two types, one group was induced by the antithrombotic therapies for CHD, and the other group was caused by some concurrent diseases or combined non-antithrombotic medications. There were different medical decisions and prognoses according to the causes in different groups. Conclusion The treatment strategies of CHD will be considered in the CHD patients with thrombocytopenia. Identifying thrombocytopenia by monitoring the platelet counts in early stage, finding out the causes quickly and providing proper treatments are the key for the prognosis of the patients.