AIM:To explore the role of survivin-2B in the process of tumor cell apoptosis .METHODS:The survivin-2B gene was cloned into pcDNA3.1 vector and the recombinant plasmid pcDNA3.1-survivin-2B was obtained.Hu-man breast cancer MCF7 cells were transfected with pcDNA3.1 and pcDNA3.1-survivin-2B using Lipofectamine 2000.The cell cycle was determined by propidium iodide staining , and the apoptosis was detected by annexin V/7-AAD staining 48 h after transfection.Meanwhile, tatal RNA was extrated and multiplex polymerase chain reaction based on GenomeLab GeXP Genetic Analysis System was performed to detect the expression of 21 tumor-related genes .RESULTS: Flow cytometry analysis indicated that over-expression of survivin-2B promoted the apoptosis and cell cycle arrest of MCF 7 cells.Compared with control group , totally 10 differential expressed genes were related to the over-expressed survivin-2B, among which 2 were up-regulated and 8 were down-regulated. The expression of aldehyde dehydrogenase 4 family member A1 (ALDH4A1) was 48%down-regulated, and the expression of protein regulator of cytokinesis 1 (PRC1) was 1.08 folds up-regulated.CONCLUSION:Survivin-2B induces the expression changes of some tumor-related genes, which results in the apoptosis and G 2/M arrest of MCF7 cells.

Objective To investigate the anti-atherosclerosis effect of apocynum venetum(AV)by observing the influence of AV extract on early inflammatory factor TNF-αexpression.Methods Human U937 monocytes were differentiated to macrophages by the phorbol myristate acetate(PMA)induction and acted with 100 mg/L ox-LDL to form the foam cells for establishing the early atherosclerosis model(ox-LDL group).The different concentrations(0.2,0.4,0.8 mg/L)of AV were added to co-culture for 48 h (AV1,AV2,AV3 groups).The expression level of TNF-αin the supernate was detected by ELISA and RT-PCR respectively.Re-sults Compared with control group,the expression level of TNF-αin the ox-LDL group was significantly increased,the expression level of TNF-αin various AV medication groups(AV1,AV2,AV3 groups)was significantly decreased compared with the ox-LDL group(P<0.05).The AV concentration increase was negatively correlated with the TNF-αexpression level(P<0.05).Conclusion TNF-αis an important inflammatory factor in early atherosclerosis,AV could play the anti-atherosclerosis role by inhibiting the inflammatory factors.

OBJECTIVETo establish a method of culturing endothelial cells (EC) from human vascular malformation.

METHODSVenous malformation specimens obtained from the patient undergoing oral surgery were plated into glass, plastic and gelatin-coated dishes. Pure cultures of human vascular malformation endothelial cells (VMEC) were isolated by the ways of discarding the tissues at early stage of primary culture, scraping and trypsinizing. Morphological characteristics were studied under phase-contrast microscope and electron microscope (EM), and determined by immunohistochemistry.

RESULTSThe cells were pure and could be maintained in culture up to 4 approximately 5 passages, or 40 approximately 50 days. VMEC formed contact-inhibited "cobblestone" monolayer on glass and plastic, and capillary-like "tubes" on gelatin. EM revealed that there were Weibel-Palade bodies in the culture cells. The cells showed positive staining for CD(34), vWF, and negative for alpha-SMA.

CONCLUSIONSThe culture technique for growing VMEC has been established. And these cells can provide a useful tool for studying biological characteristics of human vascular malformation in vitro.

Objective To investigate the effects of thyroid hormone on extremely severe traumatic brain injury (TBI).Methods A retrospective case-control study was conducted to analyze the treatment of 105 patients with extremely severe TBI admitted from July 2010 to April 2014.There were 79 males and 26 females,with an average age of 32.9 years.The patients were divided into conventional treatment group (Group A,35 cases),conventional treatment ± thyroxine treatment group (Group B,35 cases) and thyroxine treatment group after the condition that thyroxine level was low (Group C,35 cases) according to the random number table method.The incidence of low T3 and T4,incidence of hypotension,the dosage of vasoactive drugs,function evaluation of liver and kidney damage,Glasgow outcome scale (GOS),and acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) within 20 days after admission,and mortality rate within 30 days after admission were compared and analyzed.Results Within 20 days after admission,the rates of low thyroxine levels and hypotension of the Group B (22.9％,77.1％) were significantly lower than those in the other two groups (Group A:40％,100％;Group C:37％,100％) (all P ＜ 0.05).The doses of dopamine and norepinephrine in Group B was significantly lower than the other two groups and the combination rate of vasopressors in Group B was significantly lower than the other two groups (P ＜ 0.05),while there was no significant difference between Group A and Group C (P ＞ 0.05).The corresponding data in Group A and Group C had no statistically significant difference (P ＞ 0.05).The liver and renal dysfunction rates of Group B (29％,31％) were significantly lower than those of the other two groups (Group A:49％,51％;Group C:43％,51％) (all P ＜ 0.05).The corresponding data in Group A and Group C had no statistically significant difference (P ＞ 0.05).GOS in Group B [(4.8 ± 1.9) points] was significantly higher than that in Group A [(3.3 ± 0.2) points] (all P ＜ 0.05) within 30 days after admission and significantly higher than that of itself at the beginning [(3.6 ± 1.1) points] (P ＜ 0.05).The APACHE Ⅱ in Group A was significantly higher than those in other two groups as well as that in Group A at admission (P ＜ 0.05).Mortality rates in Group B (31％) and Group C (29％) were significantly lower than that in Group A (69％) within 30 days after admission (P ＜ 0.05).Conclusions Thyroxine can reduce the incidence of hypotension,liver and kidney injury rate in extremely severe TBI.Prevention is better than the supplementary treatment after severe TBI.Thyroxine can also reduce the mortality of extremely severe TBI within 30 days after admission.

Objective To investigate the clinical characteristics and the experience of multi-disciplinary team (MDT) on recurrence of primary hyperoxaluria (PH) type I after renal transplantation. Methods One case presenting with unexplained rapid decline of renal allograft function after allogeneic renal transplantation was discussed by MDT. The role of MDT in diagnosing rare hereditary diseases and improving the long-term survival of renal transplant recipients was summarized. Results After MDT consultation, the patient was diagnosed with recurrence of PH type I. Routine immunosuppressive regimen was initiated after the exclusion of rejection. The patient was instructed to drink a large quantity of water, and given with high-quality protein and low-phosphorus diet, vitamin B6, calcium and other conservative therapies to actively prevent and treat postoperative complications. The deterioration of renal graft function was delayed. Nevertheless, regular hemodialysis was resumed at 5 months after renal transplantation until the submission date of this manuscript. Conclusions Recurrence of PH type I after renal transplantation is relatively rare. The main clinical manifestations are recurrent kidney stones and decreased renal function with multiple complications and poor prognosis. The condition of the patient is consulted by MDT for confirming the diagnosis, determining the optimal treatment scheme, delaying the progression and improving the clinical prognosis.

Optimizing the drug price management mechanism and improving the availability and affordability of drugs are important in deepening the medical and health reform. The price of drugs in the United States has always been higher than the world average. The price of drugs, the total expenditure on drugs and the personal burden of patients have shown an increasing trend. By exploring the causes of high drug prices in the United States, the author found that there were four main reasons for the current situation of drug prices in the United States, including the interests of enterprises, the limited competition mechanism of the US drug market, relatively insufficient market bargaining power of the US payers, and opaque mechanism of price formation.Firstly, pharmaceutical companies try to achieve their interests by raising drug prices. Secondly, the price formation mechanism of the United States drug market is affected by the price strategy of pharmaceutical companies, and government policies also indirectly affect the role of the market. Thirdly, the payers in the United States are relatively scattered, so that the market bargaining power is relatively insufficient.Fourthly, due to the numerous drug circulation links and stakeholders, the drug price formation mechanism is opaque and lacks supervision. Therefore, when strengthening drug price management in China, we should build a coordination mechanism between the government and the market on the basis of the existing basic economic system and drug management mechanism, establish the strategic purchase and negotiation position of medical insurance for drugs, enhance the transparency of drug circulation and trading, and establish a scientific pricing system. It is also important to promote drug innovation and ensure drug quality.

Objective To explore the efficacy of repetitive peripheral magnetic stimulation(rPMS)in patients with lumbar disc herniation(LDH).Methods From March 2023 to March 2024,60 LDH patients were recruited in the inpatient or outpatient department of the rehabilitation department of a tertiary hospital.All patients were randomly assigned to the rPMS group or the conventional group,30 cases in each group.Both groups received routine physical therapy,and the rPMS group was treated with rPMS on this basis.VAS,JOA,and neurophysiological tests were performed before intervention and 2 weeks after intervention.Results The VAS and JOA scores of the two groups were significantly lower than those before treatment(P<0.05).Compared with the conventional group,the VAS and JOA scores of the rPMS group were significantly lower(P<0.05).Compared before and after treat-ment,the neuroelectrophysiological examination of the rPMS group was significantly improved(P<0.05).After 2 weeks of treatment,the tibial nerve motor conduction velocity,H reflex latency and IP peak in the conventional group were significantly faster than those before treatment(P<0.05).After 2 weeks of treatment,compared with the conventional group,there were significant differences in tibial nerve motor conduction velocity,peroneal nerve motor conduction velocity,superficial peroneal nerve sensory conduction velocity,sural nerve sensory conduction velocity,H reflex latency and IP peak(P<0.05).Conclusion rPMS can significantly improve and restore pain and nerve injury in patients with LDH.rPMS can be used as an effective adjuvant therapy.

Objective To study the effects of different preservation methods on the isolated liver injury and regeneration.Methods Twelve Sprague-Dawley rats were randomly assigned to two groups:static cold storage (SCS) and hypothermic machine perfusion (HMP) (n =6 each).Histidinetryptophan-ketoglutarate solution (HTK) was used as preservation solution.The graft was preserved in HTK (4 C) for 6 h in SCS group,and that was perfused using HTK in HMP group.Liver tissue was obtained and fixed in 10％ neutral formalin for immunohistochemistry.The fresh liver tissue was collected and stored in-80℃ for RT-PCR and Western blotting analyses.Results Compared to SCS,HMP improved liver regeneration ability in terms of PCNA and ki67 detection and promoted cell proliferation in PCR levels (Cdc25A,cdk1,cdk6) and Western blotting levels (Cyelin D1,Cyclin E1).Conclusion HMP is superior to SCS in liver regeneration,and expected to be the ideal method for preservation of donor liver.

Objective:To explore the role of interleukin-18 (IL-18) in the pathogenesis of dermatomyositis (DM) associated with positive anti-melanoma differentiation-associated gene 5 antibodies(MDA5-DM).Methods:Twenty-eight cases of MDA5-DM in the department of rheumatology and immunology, the first hospital of Nanjing medical university and the first affiliated hospital od Wannan medical colledge from August 2018 to December 2011 were included in this study, comprising 15 cases with combined rapidly progressive interstitial pneumonia (RPILD) and 13 cases without RPILD (nonRPILD). Additionally, 28 cases of antisynthetase syndrome (ASS) and 28 healthy volunteers (HC) were included for comparison. Clinical, laboratory, and imaging data were collected for both the DM and ASS groups. Serum IL-18 levels were measured using ELISA. Independent t test, Mann-whitney U test, χ2 test and Fisher′s exact probability method were used for analysis. Results:Significant differences were observed in LDH, hydroxybutyrate dehydrogenase (HBDH), ESR, CRP, serum ferritin (SFE), and IL-18 levels between the MDA5-DM group, the ASS group and the HC group ( F=46.65, 43.19, 31.28, 23.94, 30.94, 49.44, all P<0.001). Additionally, lymphocyte counts and hemoglobin levels differed significantly among the three groups( F=35.26, P<0.001; F=18.59, P<0.001). MDA5-DM patients exhibited higher incidences of Gottron′s sign, helitrope rash, periungual erythema, skin ulcers, and RPILD compared to ASS patients ( χ2=20.96, P<0.001; χ2=5.85, P=0.016; χ2=13.69, P<0.001; χ2=9.16, P=0.002; χ2=4.79, P=0.029). However, the incidence of mechanic′s hand was lower in MDA5-DM patients ( χ2=3.90, P=0.048). The level of IL-18 significantly decreased in MDA5-DM after treatment[(104.28(71.96,151.10)pg/ml vs. 78.30(56.20, 94.80)pg/ml, =2.27, P=0.023)]. Similar reductions were observed in the ASS group[(72.30(61.39, 95.94)pg/ml vs. 45.30(29.00,84.10)pg/ml, Z=2.691, P=0.007]. The IL-18 level changes in the MDA5-DM combined with RPILD group were not statistically significant [99.49 (77.65, 130.87)pg/ml vs. 89.40(54.80, 120.20)pg/ml, Z=0.65, P=0.515]. In the MDA5-DM survival group, IL-18 levels decreased significantly after treatment [59.45(53.58, 81.63)pg/ml vs. 106.37(83.62, 152.07)pg/ml, Z=2.80, P=0.005], while the changes in the IL-18 levels of patients in the MDA5-DM death group were not statistically significant [99.49(56.70, 140.15)pg/ml vs. 94.80(71.40, 155.45)pg/ml, Z=1.75, P=0.080]. Conclusion:MDA5-DM patients are different from the ASS patients in clinical manifestations and indicators involved in laboratory tests. The expression level of IL-18 tends to increase during the active phase of MDA5-DM and ASS, and decrease with remission of the disease. MDA5-DM may play an important role in the pathogenesis, and persistent high level of IL-18 is responsible for RPILD and death of MDA5-DM. Sustained high level of IL-18 can be used as a potential biomarker for the estimating development of MDA5-DM into RPILD.

The anterior cruciate ligament (ACL) injury is a common sports injury that has a significant impact on knee function and patients′ mobility. With the popularity of national fitness campaign in China, the incidence of ACL injury is increasing year by year. Currently, there still lacks clinical standards or guidelines on how to choose appropriate treatment methods, surgical plans and rehabilitation protocols for ACL injury. In order to timely reflect the new treatment concept of ACL injury, standardize its diagnosis and treatment and improve the curative effect, the Sports Medicine Society of Chinese Research Hospital Association and the Editorial Board of Chinese Journal of Trauma organized domestic orthopedic and sports medicine experts to formulate the "clinical evidence-based guideline for the diagnosis and treatment of anterior cruciate ligament injury (2022 version)" based on the level of evidence-based medicine and in compliance with the principle of scientificity, practicability and advancement. The present guideline includes 12 recommendations for the diagnosis, treatment and rehabilitation of ACL injury in order to provide guidance and assistance for the clinical diagnosis and treatment of ACL injury in China.