Clustered regularly interspaced short palindromic repeats（CRISPR）/CRISPR associated nuclease 9 （CRISPR/Cas9） is a self-defense system found in bacteria and archaea that enables targeted gene editing based on the principle. Due to its universality， efficiency， and simplicity， CRISPR/Cas9 has been applied in the pathological mechanism and prevention and treatment of diseases in many fields. Cerebrovascular diseases and central nervous system diseases seriously endanger human health. Stroke is related to genetics， unhealthy living habits， chronic diseases， and other factors. The brain tissue structure is complex and the cell types are diverse. It is difficult for a universal gene editing platform to study target genes safely， specifically， and efficiently. Scholars have continuously improved and optimized gene editing technology， explored the potential and research methods of gene editing technology， and promoted the research process of brain science. After a brief introduction to the mechanism of CRISPR/Cas9， this paper mainly summarized the optimization of the system in the fields of cerebral science including delivery methods， adeno-associated virus assembly， and new nanomaterials. Its application in cerebrovascular research including vascular homeostasis， microglial homeostasis， angiogenesis， blood-brain barrier， and drug screening was also summarized. Finally， this paper prospected the development of CRISPR/Cas9 in traditional Chinese medicine， hoping to provide references for related research design.

Objective:To study the changes of early (i.e., within post injury day (PID) 14) coagulation function in patients with severe burns.Methods:A retrospective case series study was conducted. From December 2018 to December 2019, 50 severe burn patients who met the inclusion criteria were admitted to Guangzhou Red Cross Hospital of Jinan University. According to the severity of burns, the patients were divided into severe burn group (17 cases, including 12 males and 5 females) and extremely severe burn group (33 cases, including 26 males and 7 females). The platelet count (PLT), and conventional coagulation indexe and thromboelastogram index levels of patients were collected at admission, post injury hour (PIH) 48 and 72, and on PID 7 and 14. The conventional coagulation indexes included prothrombin time (PT), thrombin time (TT), activated partial prothrombin time (APTT), and fibrinogen (FIB) and D-dimer levels. The thromboelastogram indexes included coagulation angle (i.e., α angle), coagulation composite index (CI), MA value, R value, and K value (reflecting maximum amplitude, coagulation reaction time, and blood agglutination time, respectively). Data were statistically analyzed with independent sample t-test, Wilcoxon rank sum test, and chi-square test. Verification of the mixed effect model was performed on each index data of patients in the two groups, while the repeated measures analysis of variance was performed on PLT. Pearson correlation analysis or Spearman correlation analysis were performed to analyze the correlation between the thromboelastogram index data (except CI) and the PLT and conventional coagulation index data, respectively. Results:At admission, PIH 48 and 72, and on PID 7 and 14, PLT of patients in severe burn group were (203±91), (148±70), (123±63), (203±62), (402±140)×10 9/L, respectively, PLT of patients in extremely severe burn group were (235±116), (145±71), (109±52), (235±106), (455±138)×10 9/L, respectively. In overall comparison, only the difference of the main effect of time factor was statistically significant ( F=92.55, P<0.05). In severe burn group, statistically significant differences were only identified in comparison of patients' PLT between PID 7 and the adjacent two time points (at PIH 72 and on PID 14, with both P values <0.05). The differences in PLT of patients between all the adjacent time points in extremely severe burn group were statistically significant ( P<0.05). In the overall comparison of PT, TT, and FIB level of patients in the two groups at each time point, only the difference of main effect of time factor was statistically significant (with F values of 6.04, 8.45, and 32.90, respectively, all P values <0.05), and APTT and FIB level of patients in extremely severe burn group within PID 14 were higher than those in severe burn group. There were statistically significant differences in MA value, α angle, K value, and CI of patients in the two groups at each time point (with F values of 18.82, 11.38, 9.11, and 9.42, respectively, all P values <0.05). MA value was moderately correlated with PLT ( r=0.69, P<0.05), weakly correlated with TT and FIB level (with r values of -0.29 and 0.30 respectively, P<0.05), and very weakly correlated with D-dimer level ( r=-0.15, P<0.05); α angle was moderately correlated with PLT ( r=0.58, P<0.05), and weakly correlated with FIB level and TT (with r values of 0.26 and -0.29, respectively, P<0.05); R value was weakly correlated with APTT and FIB level (with r values of 0.24 and 0.31, respectively, P<0.05), and very weakly correlated with PT and TT (with r values of 0.16 and 0.14, respectively, P<0.05); K value was moderately correlated with PLT ( r=-0.59, P<0.05), and weakly correlated with FIB and TT (with r values of -0.29 and 0.32, respectively, P<0.05), and very weakly correlated with D-dimer level ( r=-0.15, P<0.05). Conclusions:Severe burn patients are already characterized with coagulation function changes in early stage, including insufficiency of coagulation function, enhanced platelet aggregation ability and enhanced FIB function. There is a certain correlation between conventional coagulation indexes and thromboelastogram indexes, but they cannot replace each other.

ObjectiveTo observe the protective effect of Shenlian prescription on acute lung injury induced by particulate matter （PM） exposure in rats and explore the mechanism. MethodFifty male SD rats were randomly divided into the control group， model group， Shenlian low-dose group （4.32 g·kg^-1）， Shenlian high-dose group （8.64 g·kg^-1）， and roflumilast group （3.46 mg·kg^-1）， with 10 in each group. Pre-administration with drugs by gavage was performed for one week. On the 8^th and 11^th days， the control group was instilled with normal saline in the trachea and the other groups with PM suspension to establish a rat model of acute lung injury induced by PM exposure. After modeling， drugs were given continuously until the end of the experiment. Forty-eight hours after the last exposure， the lung function of rats was detected. Then the rats were sacrificed and the lung morphological changes and pathological changes by hematoxylin-eosin （HE） staining were observed. CD68 expression in lung was detected by immunohistochemistry， and the levels of lung injury markers surfactant protein A （SP-A） and Clara cell protein16 （CC16） in serum were detected by enzyme-linked immunosorbent assay （ELISA）. The mRNA expression of interleukin-1α （IL-1α）， IL-6， IL-18， and monocyte chemoattractant protein-1 （MCP-1） in lung tissue was measured by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. ResultCompared with those in the control group， the rats in the model group had decreased lung function and obvious structural damage of lung tissue， PM deposition， and infiltration of CD68 positive cells. The expressions of IL-1α， IL-6， IL-18， and MCP-1 in lung tissue were increased （P<0.01）. Compared with the model group， Shenlian prescription low and high doses restored the rats' lung function injury（P<0.05，P<0.01）， improved lung morphological and pathological structure， and reduced PM deposition. Infiltration of CD68 positive cells in lung was not significantly decreased. The levels of inflammatory factors IL-1α， IL -6， IL-18， and MCP-1 in lung were lowered （P<0.01）. ConclusionShenlian prescription could protect the rats' lung injury caused by PM exposure， improve lung morphology， and reduce PM deposition and inflammatory factor expression.

Objective:To understand the diagnostic value of tuberculosis (TB) pathogenic detection methods (TPDM) in pathological tissue for TB.Methods:A retrospective study was conducted with 190 pathological specimens from different tissues suspected with TB from Third People′s Hospital of Shenzhen during May 2016 and May 2019. Specimens were divided into four groups according to histomorphology: group one, necrotizing granulomatous inflammation (109 cases); group two, non-necrotic granulomatous inflammation (20 cases); group three, non-granulomatous inflammation (45 cases); group four, non-tuberculous lesions (16 cases). The positive rates of each TPDM among specimens from four groups were compared. The positive rates of all TPDM for specimens from group one were compared. Meanwhile, the influence of antituberculosis treatment course on the TPDM was analyzed. Chi-square test or Fisher′s exact test was used for statistical analysis.Results:The positive rates of Ziehl-Neelsen acid-fast staining among the four groups were 17.4%(19/109), 5.0%(1/20), 4.4%(2/45) and 0(0/16), respectively. The positive rates of Mycobacterium tuberculosis (MTB) complex culture were 32.0%(32/100), 4/19, 4.8%(2/42) and 0(0/16), respectively. The positive rates of Mycobacterium tuberculosis/rifampin resistance real-time quantitative nucleic acid amplification detection system (Xpert MTB/RIF) were 74.3%(81/109), 15.0%(3/20), 13.3%(6/45) and 0(0/16), respectively. The positive rates of fluorescent quantitative polymerase chain reaction (FQ-PCR) were 63.0%(58/92), 0(0/15), 2.6%(1/38) and 0(0/10), respectively. The positive rates of simultaneous amplification and testing (SAT) were 32.4%(24/74), 0(0/10), 0(0/15) and 0(0/10), respectively. The differences of each TPDM among four groups were all statistically significant (all P<0.05). The positive rate of Xpert MTB/RIF in group one specimens was significantly higher than those of acid-fast staining, MTB culture and SAT ( χ2=71.016, 37.162 and 35.679, respectively, all P<0.01), while the difference was not statistically significant when compared with FQ-PCR ( χ2=2.517, P=0.112). The positive rate of combined TPDM (85.3%(93/109)) was significantly higher than Xpert MTB/RIF(74.3%(81/109)) ( χ2=4.100, P=0.043). The positive rates of acid-fast staining group 1A (anti-tuberculosis treatment course was less than one month) and group 1B (anti-tuberculosis treatment course was longer than one month) were 14.3%(7/49) and 20.0% (12/60), respectively ( χ2=0.612, P=0.434); those of MTB culture were 48.9% (22/45) and 18.2% (10/55), respectively ( χ2=10.721, P=0.001); those of Xpert MTB/RIF were 69.4%(34/49) and 78.3%(47/60), respectively ( χ2=1.131, P=0.287); those of FQ-PCR were 55.0%(22/40) and 69.2%(36/52), respectively ( χ2=1.965, P=0.161); those of SAT were 43.3%(13/30) and 25.0%(11/44), respectively ( χ2=2.736, P=0.098). Conclusions:The results of TPDM correlate closely with the typical histomorphological features of tuberculosis. Xpert MTB/RIF possesses significantly higher sensitivity than any other single TPDM, and is not attenuated by early anti-tuberculosis treatment. Combined TPDM could significantly improve the sensitivity of TB pathogenic detection, which is suggested to be applied when the tissue specimen is sufficient.

During the epidemic of coronavirus disease 2019 (COVID-19), the infection of the elderly population will bring great challenges to clinical diagnosis and treatment, outcome and management.Combined with the characteristics of anesthesia and the pathophysiological characteristics of COVID-19 on lung function impairment in elderly patients, Chinese Society of Anesthesiology formulated the " Recommendations for anesthesia management and infection control in elderly patients with COVID-19″. This recommendation expounds preoperative visit and infection control, anesthesia management protocol, anesthesia monitoring, anesthesia induction/endotracheal intubation, anesthesia maintenance and infection control, intraoperative lung protection strategy, anti-stress and anti-inflammatory management, hemodynamic optimization, infection control during emergence from anesthesia, and postoperative analgesia in elderly patients with COVID-19, and provides the reference for the safe and effective implementation of anesthesia management in elderly patients during the prevention and control of COVID-19 epidemic.

Objective: To analyze the causes of complication of early acute kidney injury (AKI) in four severely burned patients, and to explore the related treatment methods. Methods: The clinical data of 4 patients with severe burn complicated with early AKI admitted to Guangzhou Red Cross Hospital Affiliated to Medical College of Jinan University (hereinafter referred to as our hospital) from June 2014 to December 2017 were retrospectively analyzed. All the patients were male, aged 23-33 (30±5) years old, with depth of burns ranged from deep partial-thickness to full-thickness, complicated with myofascial compartment syndrome of extremities and varying degrees of striated muscle injury, and treated in other hospitals before transfer to our hospital. The patients were numbered from small to large according to the total burn area. The total burn area of patients No. 1, 2, 3, and 4 was 10%, 80%, 90%, and 95% total body surface area respectively, their occurrence time of early AKI was 48, 11, 29, and 48 hours after injury respectively, and their time of arriving our hospital was 60, 11, 29, and 144 hours after injury respectively. Hypovolemic shock occurred in patients No. 2 and 3 at admission to our hospital. All the patients received continuous renal replacement therapy (CRRT) after admission to our hospital. Under the support of hemodynamic monitoring and organ function monitoring, the limbs complicated with myofascial compartment syndrome were incised, thorough decompression exploration was performed, and necrotic muscle tissue was removed or amputation was performed. After escharectomy and decompression of limbs, fresh granulation wounds were formed by temporarily covering wounds with Jieya dressing skin or pig skin, multiple debridements, and vacuum sealing drainage. Fresh granulation wounds and other wounds underwent staged eschar excision and shaving were covered with autologous Meek skin graft, particulate skin graft, reticular skin graft and small skin graft respectively. The treatment outcome, CRRT time, operation times, time of recovery of serum creatinine and myoglobin, length of hospital stay, and follow-up were recorded. Results: All the 4 patients were cured after transfer to our hospital. Among them, totally 5 limbs of patients No. 1 and No. 4 underwent amputation because of complication of myofascial compartment syndrome and a large amount of necrotic muscle which could not be preserved. Patients No. 1, 2, 3, and 4 were treated with CRRT for 19, 35, 14, and 25 days respectively and performed with operation for 5, 6, 10, 8 times respectively. Serum creatinine of patients No. 1, 2, 3, and 4 returned to normal on 22, 35, 37, and 48 days after transfer respectively, and their serum myoglobin returned to normal on 18, 28, 25, and 30 days after transfer respectively. Patients No. 1, 2, 3, and 4 were hospitalized for 52, 105, 148, and 156 days and discharged after basic wound healing. Follow-up for 1 to 36 months showed no abnormal renal function in 4 patients. Conclusions The early AKI in patients No. 1 and 4 was caused by rhabdomyolysis after severe burn complicated with myofascial compartment syndrome, while that of the other 2 cases were also related to hypovolemic shock and poor renal perfusion. The success rate of early AKI treatment in severely burned patients can be effectively improved by removing the causes of diseases at the same time of CRRT and actively treating burn wounds under the support of organ function and hemodynamic monitoring.

Objective To explore the feasibility and efficacy of endoscopic balloon dilation in treatment of esophageal stenosis caused by operation of congenital esophageal atresia. Methods A retrospective analysis was performed on data of 218 children with type Ⅲ esophageal atresia, who underwent surgery in Zhengzhou Children' s Hospital from January 2009 to December 2017. The occurrence of postoperative complications and efficacy of endoscopic balloon dilation in treatment of esophageal stenosis was analyzed. Results Among the 218 patients with congenital esophageal atresia, 92 were type Ⅲa and 126 were type Ⅲb. Postoperative anastomotic leakage occurred in 46 cases (21. 1％), including 29 (31. 5％) of type Ⅲa and 17 (13. 5％) of type Ⅲb. Postoperative anastomotic stenosis occurred in 53 cases (24. 3％), including 29 ( 31. 5％) of type Ⅲa and 24 ( 19. 0％) of typeⅢb. The incidence of anastomotic leakage and anastomotic stenosis in different types was significantly different (χ2=10. 383, P=0. 001; χ2=4. 497, P=0. 034). The 53 cases of anastomotic stenosis underwent 123 times of endoscopic balloon dilation, with mean time of 3. 5±1. 6, and were finally clinically recovery. No esophagus perforation occurred. Among them, 29 cases of type Ⅲa underwent 73 times with mean of 4. 0±1. 8, and 24 cases of type Ⅲb underwent 50 times with mean of 2. 5±0. 7. The difference between the two types was statistically significant (t=-4. 053, P=0. 027). Conclusion Children with type Ⅲa esophageal atresia has a higher incidence of anastomotic stenosis and leakage, and more times of esophageal dilation. Endoscopic balloon dilation is safe and effective in treatment of esophageal stenosis after surgery for patients with congenital esophageal atresia.

Objective: RET/PTC gene rearrangement can lead to aberrant activation of tyrosine kinase receptors, which is a common mutation in papillary thyroid carcinoma (PTC). This study focuses on the association of RET/PTC rearrangements with PTC clinical factors. Methods: From January 2011 to December 2013, a total of 114 patients with PTC were enrolled in this study. Clinicopathological parameters, lifestyle, and thyroid hormone levels were collected. RET/PTC rearrangements were detected by TaqMan PCR and verified by Sanger sequencing.Data were analyzed with SPSS software, including chi-square test, Fisher′s exact test, Mann-Whitney U test, Student′s t-test, and Logistic regression. Results: RET/PTC rearrangements were not found in all paracancerous normal thyroid tissues, and were detected in 23.68% (27/114) of PTC. Further analysis revealed no correlation between RET/PTC rearrangement and thyroid function, clinicopathologic parameters, and lifestyle in the total PTC group or in the subgroup of patients with concomitant diseases (including Hashimoto′s thyroiditis and nodular goiter). But in the subgroup of PTC without concomitant disease, RET/PTC rearrangement was associated with tumor multifocal (P=0.018), and RET/PTC-positive PTC patients had an increased risk of tumor multifocal (OR=5.57, 95% CI 1.39-22.33). It was also found that RET/PTC rearrangement was associated with an abnormal increase in TSH level of one month after surgery (P= 0.037). Conclusion Nodular goiter and Hashimoto ′s thyroiditis may be a confounding factor in PTC. RET/PTC rearrangement may play an important role in the occurrence of thyroid carcinoma multifocal after exclusion of this confounding factor.

Objective To evaluate the efficiency of multimodal analgesia for the patients who had the posterior lumbar spine surgery.Methods Sixty-four cases of patients who were scheduled to the posterior lumbar spine surgery were randomly divided into two groups, multimodal analgesia group (n =32) and control group(n=32).Multimodal analgesia group patients were given oxycodone acetaminophen 1 day before and the next day after operation, during the operation the patients received injiection of multimodal drugs (consisting of ropivacaine 150 mg, flurbiprofen 50 mg, phenylephrine 0.4 ml, normal saline 50 ml) around the incisions by infiltrated injection before the skin incision closed, then had controlled intravenous analgesia.In the control group, the incisions were sutured without the local infiltration analgesia.Then only had patient controlled intravenous analgesia after operati on.Visual analogue scale(VAS) ,Japanese Orthopedic Association(JOA) and Prolo lumbar function score was recorded respectively perioperatively.Results (1) VAS: the patients of multimodal analgesia group had significantly lower rest pain scores and activity pain scores at 6, 12,24 hours, first moving than the control group((3.1±1.6) ,(2.8±1.1),(2.4±0.9) ,(2.3±1.1) vs.(3.5±1.8) ,(3.4± 1.3), (3.4±0.8), (3.0± 1.5), P＜0.05).There was no significant difference at the discharge between the two groups (P＞0.05).(2)JOA:JOA scores of the patients of multimodal analgesia group were obviously higher than that of the control group at 1,3,7 days after operation (P＜0.05).There was no significant difference at the discharge between the two groups (P＞0.05).(3) Prolo lumbar function score : Prolo scores of the patients of the multimodal analgesia group were obviously higher than that of control group at 1,3,7 days after operation (P ＜0.05).There was no significant difference at the discharge between the two groups(P＞0.05).Conclusion Multimodal Analgesia is shown to safely provide excellent pain control and functional recovery.It can reduce visual analogue pain score and improve lumbar function after surgery.

OBJECTIVETo investigate the effects of unified surgical scheme for wounds on the outcome of patients with extensive deep partial-thickness to full-thickness (briefly referred to as deep) burn.

METHODSOne hundred and thirty-seven patients with extensive deep burn hospitalized from July 2007 to November 2012 underwent unified surgery according to area of deep wound (unified scheme group, US). Among them, 57 patients with deep wound area less than 51% TBSA received escharectomy or tangential excision by stages followed by autologous mesh skin grafting; 52 patients with deep wound area from 51% to 80% TBSA underwent escharectomy or tangential excision by stages followed by autologous mesh skin grafting and/or small skin grafting, or escharectomy or tangential excision followed by large sheet of allogeneic skin covering plus autologous mesh skin grafting and/or small skin grafting after the removal of allogeneic skin; 28 patients with deep wound area larger than 80% TBSA received escharectomy or tangential excision by stages followed by autologous microskin grafting plus coverage of large sheet of allogeneic skin, or escharectomy or tangential excision followed by small autologous skin grafting and/or intermingled grafting with small autologous and/or allogeneic skin. Another 120 patients with extensive deep burn hospitalized from January 2002 to June 2007 who did not receive unified surgical scheme were included as control group (C). Except for the surgical methods in group US, in 53 patients with deep wound area less than 51% TBSA in group C escharectomy or tangential excision was performed followed by autologous small skin grafting; in 40 patients with deep wound area from 51% to 80% TBSA in group C escharectomy or tangential excision was performed followed by autologous microskin grafting plus large sheet of allogeneic skin covering, or escharectomy or tangential excision followed by large sheet of allogeneic skin embedded with stamp-like autologous skin; in 27 patients with deep wound area larger than 80% TBSA in group C escharectomy or tangential excision was performed followed by covering with large sheet of allogeneic skin embedded with stamp-like autologous skin without intermingled grafting with small autologous and allogeneic skin in group US. In group US, escharectomy of full-thickness wound in extremities was performed with the use of tourniquet in every patient; saline containing adrenaline was subcutaneously injected when performing escharectomy or tangential excision over the trunk and skin excision; normal skin and healed superficial-thickness wound were used as donor sites for several times of skin excision. The baseline condition of patients and their treatment in the aspects of fluid resuscitation, nutrition support, anti-inflammation, and organ function support were similar between the two groups. The mortality and incidence of complications of all patients and wound healing time and times of surgery of healed patients were compared between the two groups. Data were processed with independent sample t test, Mann-Whitney U test, and Fisher's exact test.

RESULTS(1) Both the mortality and the incidence of complications of patients with deep wound area less than 51% TBSA in group US were 0, which were close to those of group C (with P values above 0.05). The number of times of surgery of healed patients with deep wound area less than 51% TBSA in group US was 2.4 ± 0.9, which was obviously fewer than that of group C (3.5 ± 1.8, U=-5.085, P<0.001), but with wound healing time close to that of group C (U=-1.480, P>0.05). (2) Both the mortality and the incidence of complications of patients with deep wound area from 51% to 80% TBSA in group US were 0, which were significantly lower than those of group C [both as 20.0% (8/40), with P values below 0.01]. The number of times of surgery and wound healing time of healed patients with deep wound area from 51% to 80% TBSA in group US were respectively 3.0 ± 1.0 and (43 ± 13) d, which were obviously fewer or shorter than those in group C [4.2 ± 2.3 and (61 ± 34) d, with U values respectively -2.491 and -2.186, P values below 0.05]. (3) Both the mortality and the incidence of complications of patients with deep wound area larger than 80% TBSA in group US were 25.0% (7/28), which were close to those of group C [both as 25.9% (7/27), with P values above 0.05]. The number of times of surgery and wound healing time of healed patients with deep wound area larger than 80% TBSA in group US were close to those of group C (with U values respectively -0.276 and -0.369, P values above 0.05).

CONCLUSIONSUnified surgical scheme can indirectly decrease the mortality and the incidence of complications of burn patients with deep wound area from 51% to 80% TBSA; it can reduce times of surgery of healed patients of this type and shorten their wound healing time.

Burns ; surgery ; Debridement ; methods ; Extremities ; Humans ; Severity of Illness Index ; Skin ; pathology ; Skin Transplantation ; Transplantation, Autologous ; Treatment Outcome ; Wound Healing