CRISPR/Cas9-based therapeutics face significant challenges in penetrating the dense microenvironment of solid tumors, resulting in insufficient gene editing and compromised treatment efficacy. Current nanostrategies, which mainly focus on the paracellular pathway attempted to improve gene editing performance, whereas their efficiency remains uneven in the heterogenous extracellular matrix. Here, the nanoCRISPR system is prepared with self-cascading mechanisms for gene editing-mediated robust apoptosis and transcellular penetration. NanoCRISPR unlocks its self-cascade capability within the matrix metallopeptidase 2-enriched tumor microenvironment, initiating the transcellular penetration. By facilitating cellular uptake, nanoCRISPR triggers robust apoptosis in edited malignancies, promoting further transcellular penetration and amplifying gene editing in neighboring tumor cells. Benefiting from self-cascade between robust apoptosis and transcellular penetration, nanoCRISPR demonstrates continuous gene transfection/tumor killing performance (transfection/apoptosis efficiency: 1st round: 85%/84.2%; 2nd round: 48%/27%) and homogeneous penetration. In xenograft tumor-bearing mice, nanoCRISPR treatment achieves remarkable anti-tumor efficacy (∼83%) and significant survival benefits with minimal toxicity. This strategy presents a promising paradigm emphasizing transcellular penetration to enhance the effectiveness of CRISPR-based antitumor therapeutics.

Objective To investigate the relationship of miRNA gene polymorphisms with blood pressure(BP)responses to the sodium and potassium diet intervention.Methods In 2004,we recruited 514 participants from 124 families in seven villages of Baoji,Shaanxi Province,China.All subjects were given a three-day normal diet,followed by a seven-day low-salt diet,a seven-day high-salt diet,and finally a seven-day high-salt and potassium supplementation.A total of 19 miRNA single nucleotide polymorphisms(SNPs)were selected for analysis.Results Throughout the sodium-potassium dietary intervention,the BP of the subjects fluctuated across all phases,showing a decrease during the low-salt period and an increase during the high-salt period,followed by a reduction in BP subsequent to potassium supplementation during the high-salt diet.MiR-210-3p SNP rs 12364149 was significantly associated with systolic BP(SBP),diastolic BP(DBP)and mean arterial pressure(MAP)responses to low-salt diet.MiR-4638-3p SNP rs6601178 was significantly associated with SBP while miR-26b-3p SNP rs115254818 was significantly associated with MAP responses to low-salt intervention.In addition,miR-26b-3p SNP rs115254818 was significantly correlated with SBP,DBP and MAP responses to high-salt intervention.MiR-1307-5p SNPs rs1 1191676 and rs2292807 were associated with SBP and MAP responses to high-salt diet.MiR-4638-3p SNP rs6601178,miR-210-3p SNP rs12364149,miR-382-5p SNP rs4906032 and rs4143957 were significantly associated with SBP response to high-salt diet.In addition,miR-26b-3p SNP rs115254818 was significantly associated with SBP,DBP and MAP responses to potassium supplementation.MiR-1307-5p SNPs rs11191676,rs2292807,and miR-19a-3p SNP rs4284505 were significantly associated with SBP responses to high-salt and potassium supplementation.Conclusion miRNA gene polymorphisms are associated with BP response to sodium and potassium,suggesting that miRNA genes may be involved in the pathophysiological process of salt sensitivity and potassium sensitivity.

ObjectiveTo explore the evolution principles of symptoms including deficiency， phlegm and blood stasis， and of the functional near-infrared spectroscopy （fNIRS） cerebral hemodynamic characteristics at various stages in patients of Alzheimer's disease. MethodsA total of 497 patients with complaint of memory loss were included， and were divided into subjective cognitive decline （SCD） group （198 participants）， mild cognitive impairment （MCI） group （228 participants） and dementia （AD） group （71 participants）. Neuropsychological evaluation， traditional Chinese medicine （TCM） syndrome investigation， and fNIRS data collection of prefrontal cortex were performed in each group. Descriptive statistics were used to analyze the distribution of TCM syndromes and the difference of TCM syndrome scores in each group； logistic regression was used to analyze the influence of TCM syndromes on the incidence of the patients； association rules were used to analyze the TCM syndromes of the patients； the hemodynamic characteristics of fNIRS in the prefrontal cortex of each group were compared. ResultsKidney essence deficiency syndrome was the dominant syndrome in all stages of AD. There were statistically significant differences in the distribution frequency of kidney essence deficiency， phlegm turbidity obstructing orifices， blood stasis obstructing collaterals， qi and blood deficiency， heat toxin in the interior， and fu-organ stagnation and turbidity retention syndromes among the three groups （P＜0.01）， and the scores of kidney essence deficiency syndrome among the three groups were statistically significant （P＜0.01）. Logistic regression analysis showed that kidney essence deficiency， and qi and blood deficiency syndromes were the main risk factors for the SCD group （P＜0.05）， phlegm turbidity obstructing orifices syndrome was the main risk factor for the MCI group （P＜0.05）， and heat toxin in the interior， and fu-organ stagnation and turbidity retention syndromes were the main risk factors for the AD group （P＜0.05）. The association rule analysis showed that the combination of kidney essence deficiency plus phlegm turbidity obstructing orifices had the highest support （33.33%） in the SCD group， and the combination of kidney essence deficiency plus blood stasis obstructing collaterals had the highest support （32.90% and 52.13%） in both the MCI and AD group. The prefrontal fNIRS results showed that the mean ∆HbO₂ concentration in the left dorsolateral prefrontal cortex （LDLPFC） decreased sequentially among the three groups （P＜0.05）， and the mean ∆HbO₂ concentration in the LDLPFC was negatively correlated with the MoCA score among the three groups （r = -0.142， P＜0.05）. Further analysis showed that the mean ∆HbO₂ concentration in the LDLPFC of patients with kidney essence deficiency syndrome were statistically significant differences among the three groups （P＜0.05）. ConclusionKidney deficiency is the basis of the pathogenesis of AD， and the key brain area damaged is the LDLPFC. Turbid pathogens such as phlegm and blood stasis are the pathological factors that aggravate the disease， and the syndromes of AD show the evolution law of deficiency and excess as “kidney deficiency→phlegm turbidity→blood stasis→turbid toxin”. The changes in prefrontal hemodynamics based on fNIRS are consistent with the changes in the characteristics of symptoms， which can be used to assess the degree of cognitive impairment in AD patients.

【Objective】 Corin, a transmembrane serine protease that can cleave atrial natriuretic peptide precursor (pro-ANP) into atrial natriuretic peptide with smaller bioactive molecules, participates in the pathophysiological process of hypertension and cardiac hypertrophy. The purpose of this study was to explore the relationship of Corin gene variation with blood pressure responses to sodium and potassium dietary interventions. 【Methods】 In 2004, we recruited 514 participants from 124 families in 7 villages of Baoji, Shaanxi Province, China. All the subjects received a 3-day normal diet, a 7-day low-salt diet, a 7-day high-salt diet, and finally a 7-day high-salt and potassium supplementation. Fifteen single nucleotide polymorphisms (SNPs) of Corin gene were selected for final analysis. 【Results】 SNPs rs12509275 were significantly associated with diastolic blood pressure (DBP) response to low-salt diet, while rs3749584 was associated with pulse pressure (PP) response to low-salt diet.SNP rs3749584 and rs10517195 were significantly associated with PP response to high-salt diet. In addition,rs17654278 were significantly associated with systolic blood pressure (SBP) response to high-salt and potassium supplementation, rs2271037 was significantly correlated with DBP responses to high-salt and potassium supplementation, and rs4695253, rs12509275, rs2351783, rs36090894 were significantly associated with PP response to high-salt and potassium supplementation. 【Conclusion】 Corin gene polymorphisms were associated with blood pressure response to sodium and potassium, suggesting that Corin gene may be involved in pathophysiological process of salt sensitivity and potassium sensitivity.

Objective To conduct an empirical analysis of constitutes and changes of type 2 diabetes patients′ hospitalization expenses and find out the differences and influencing factors so as to provide hospitals and related departments with the decision basis of controlling medical costs, arranging medical resources and optimizing systems.Methods Information of type 2 diabetes patients who had been admitted to a certain first-class grade one hospital in Ningxia from 2013 to 2016 were collected and categorized.Hospitalization costs and influencing factors of the patients were comprehensively analyzed through descriptive statistics, variance analysis and regression analysis.Results The medicine and examination costs as the constitutes of hospitalization expenses were too high.Gender, length of stay, age and diabetic complications were main contributory factors of type 2 diabetes patients′ hospitalization expenses.Conclusion The costs of hospitalization of patients with type 2 diabetes are unreasonable and need to optimize.Type 2 diabetes patients′ hospitalization expenses should be effectively controlled and medical resources should be reasonably used through standardizing treatment behavior, shortening length of stay, intensifying publicity and education, enhancing prevention and reinforcing follow-ups, expanding health insurance coverage and upgrading its level.

BACKGROUND:Over-expression of serum interleukin-6 and interleukin-8 may be involved in tourniquet-induced limb ischemia-reperfusion injury to the lung ventilation function. OBJECTIVE:To evaluate the tourniquet effect on serum interleukin-6 and interleukin-8 levels of the rat limb within the safety time limit. METHODS: Sixty rats were randomly divided into control and experimental groups, 30 rats in each group. Rats in the control group had no ischemic preconditioning and were directly subjected to stop bleeding for 2, 3, 4 hours; rats in the experimental group were subjected to ischemic preconditioning (short-time hemostasis for several times within 1 day before ischemia-reperfusion injury), and then underwent 2-, 3-, 4-hour hemostasis at the 2nd day. At 1, 3, 7, 14 days after the recovery of limb blood flow, blood samples were extracted to detect serum interleukin-6 and interleukin-8 levels using ELISA method. RESULTS AND CONCLUSION:The levels of interleukin-6 interleukin-8 showed an increasing and decreasing trend in the two groups, which both reached the peak at the 3rd day (P< 0.05). The levels of interleukin-6 in the experimental group was significantly higher than that in the control group at 1, 3, 7, 14 days (P < 0.05), and only at 7 days, the level of interleukin-6 in the rats undergoing 3-hour hemostasis was significantly lower than that in the control group (P< 0.05). At 7 days, the level of interleukin-8 in the rats undergoing 3-hour hemostasis was lower than that in the control group, but there was no significant difference (P > 0.05), and at 3, 7, 14 days, the level of interleukin-8 in the rats undergoing 4-hour hemostasis was significantly higher than that in the control group (P< 0.05). At 1 and 3 days, the levels of interleukin-6 and interleukin-8 in the experimental group had an increasing trend with the bleeding time and ranked as folows: 2-hour hemostasis < 3-hour hemostasis < 4-hour hemostasis, and there was a significant difference; while in the control group, there was also an increasing trend in the levels of interleukin-6 and interleukin-8, but there was no statistical difference. These findings indicate that the tourniquet preconditioning treatment is preferred at 3 days after limb ischemia-reperfusion injury, when the inflammatory response was the most obvious in rats, and this treatment can dramaticaly reduce inflammatory response. Additionaly, the inflammatory become more obvious with the bleeding time.

BACKGROUND:There remains controversy that whether patel ar resurfacing in total knee arthroplasty and whether patel ar chondromalacia has influence on clinical outcomes.
OBJECTIVE:To evaluate anterior knee pain, clinical function and effects of patel ar chondromalacia on clinical outcomes after total knee arthroplasty without patel ar resurfacing for degenerative osteoarthritis.
METHODS:Clinical data of 162 patients (162 knees) with degenerative osteoarthritis undergoing total knee arthroplasty without patel ar resurfacing from June 2008 to February 2010 were retrospectively analyzed. Outerbridge classification was used for patel ar chondromalacia, Visual Analogue Scale for anterior knee pain, and the Knee Society clinical scoring system for clinical function.
RESULTS AND CONCLUSION:The incision of al patients reached stage-I healing. The patel ar chondromalacia:grade I in 18 patients, grade II in 36 patients, grade III in 62 patients, and grade IV in 35 patients. At the final fol ow-up, there were six (4.0%) patients with anterior knee pain, including four cases of mild pain and two cases of moderate pain, no severe pain. The mean Knee Society clinical scoring system scores and patel ar score were obviously elevated. Outerbridge classification did not affect the incidence of anterior knee pain after replacement (χ2=0.42, P=0.94), the Knee Society clinical scoring system score (knee score:F=1.83, P=0.14;functional score:F=0.56, P=0.64) and partel ar score (F=0.78, P=0.51). These data suggested that total knee arthroplasty without patel ar resurfacing for degenerative osteoarthritis can obtain satisfactory clinical outcomes, and the patel ar chondromalacia may not affect the clinical outcomes.

This study aimed to establish human IFN-gamma (hIFN-gamma) in vitro release assay and to apply it in diagnosis of human tuberculosis. Human IFN-gamma gene was cloned and expressed in Escherichia coli. The recombinant hIFN-gamma was purified and used as immunogen to immunize mice and rabbits respectively. Monoclonal and polyclonal antibodies were respectively developed and a sandwich ELISA was established. The heparized whole blood from 111 active tuberculosis patients and 292 clinical healthy controls were collected. The blood was stimulated with tuberculosis specific fused antigen ESAT-6/CFP-10 and the plasma was collected for IFN-gamma detection. The sensitivity for tuberculosis diagnosis was 95.5%, whereas the positive detection rate for the healthy controls was 16.7%. There was a significant difference between the patients and healthy controls (P<0.01) indicating that this assay had a high sensitivity and specificity, and thus could be promising in tuberculosis diagnosis.
Animals ; Antibodies, Monoclonal ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Interferon-gamma ; immunology ; secretion ; Mice ; Mice, Inbred BALB C ; Mycobacterium tuberculosis ; immunology ; Rabbits ; T-Lymphocytes ; immunology ; Tuberculosis ; diagnosis ; immunology