Full-speed development of the primary health care centering on community health service is key to the ongoing health reform in China. Building a new management mechanism for these grass-root health centers is an innovative approach of such a reform as carried out in Zhejiang province.The authors described the present management mechanism in the province, and focused on the specific measures and main outcomes of the nine Group-scale model as used by a district health bureau in Hanzhou since end of 2007. The paper aims at building an ideal model for group-scale management of community health centers, and improving the new city primary health care system centering on community health service.

BACKGROUND:Among many transplanted cells,adult autologous bone barrow-derived mononuclear cells have beenused in clinical practice because they are easy to be obtained,without immunological rejection and ethical disputationand other advantages.How to distinguish donor cells from receptors and observe the survival of donor cells following stem cell transplantation still trouble people.OBJECTIVE: superparamagnetic iron oxide (SPIO)particles-labeled bone marrow-derived mononuclear cells from minipigs were used to observe the feasibility of in vivo tracking with magnetic resonance imaging(MRI).DESIGN:A controlled observation experiment.SETTING:Institute of Cardiovascular Disease,Zhongda Hospital Affiliated to Southeast University.MATERIALS:This experiment was carried out in the Institute of Cardiovascular Disease,Zhongda Hospital Affiliated to Southeast University between April 2006 and August 2006.Healthy Chinese minipigs,aged 3 to 4 months,weighing from 20 to 30 kg,were provided by the Experimental Animal Center of Southeast University[SYXK(Su)2002-0012].METHODS: Autologous bone marrow-derived mononuclear cells of minipigs were isolated and cultured. Bone marrow-derived mononuclear cells in the suspension were traced with SPIO particles.Ferrum in the cells were shown by Prussian blue staining, and cell viability was evaluated by trypan blue exclusion method. Eleven minipigs used for preparation of model of myocardial infarction were divided into experimental group(n=9)and control group(n=2).By means of percutaneous left or right cervical artery or femoral artery puncturation, 1.5 to 2.0 mm balloon was used to occlude 1/3 left anterior descending branch,304 to 405 kPa,60 minutes later,ischemic preconditioning was conducted 3 tO 4 times before operation. When pig models of myocardial infarction were successful that was proved by surface electrocardiogram,bone marrow-derived mononuclear cells were percutaneously injected into coronary artery.Coronary arteriography was performed through femoral artery acupuncture at 24 hours after establishing infarction models.Suspension of bone marrow-derived mononuclear cells was perfused into coronary artery with OTW catheter.Then,the injector and OTW catheter for containing cells were rinsed with normal saline containing heparin and infused with the residual cells within 10 minutes.Non-labeled cells were perfused in 2 minipigs of control group by the same method.Postoperatively, bone marrow-derived mononuclear cells were traced by magnetic resonance and compared with Prussian blue-stained myocardial tissue sections.RESULTS: Seven minipigs of experimental group and one minipig of control group were Involved in the final analysis.One of each group was used for preparation of model of myocardial Infarction.One minipig of experimental group died from anesthetic accident before magnetic resonance.①Bone marrow-derived mononuclear cells all were nearly labeled by SPIO particles. Bone marrow-derived mononuclear cells could further proliferate in culture medium containing Fe2O3-PLL without obvious changes of cellular shape. ②T2+WI showed that 5 of 8 models of myocardial infarction presented fuzzy low-echo signal region in peripheral myocardial infarction after transplantation of labeled cells and the low-echo signal disappeared 4 weeks Iater. Ex vivo T2+WI sequence showed there was a dot-distributed low-echo signal region in the peripheral infarction region.③It was found in histological examination that 5 models(cell number over 106) had Prussian blue-positive cells,which distributed the same as those in magnetic resonance signal reducing region.CONCLUSION:SPIO particles-labeled bone marrow-derived mononuclear cells are safe and effective;T2+ WI is sensitive to tracing SPIO particles-labeled bone marrow-derived mononuclear cells;Magnetic resonance can in vivo trace SPIO particles-labeled stem cells transplanted through coronary artery,magnetic resonance signal change is related with the number of stem cells and division growth.

Gallstones and acute calculous cholecystitis are common diseases in surgery. China has entered an aging society. Elderly patients are a high incidence group of gallstones and acute calculous cholecystitis. Their surgical diagnosis and treatment strategies need to be formulated individually, and reasonable diagnosis and treatment methods should be comprehensively selected according to the patient′s condition and the medical level of the medical institution. For those who are in good physical condition and can tolerate the operation, it is recommended to give priority to surgical treatment, and the minimally invasive surgery mainly suitable for laparoscopic cholecystectomy (LC) is preferred. Open surgery is more appropriate for complicated conditions or patients whose general conditions are not suitable for laparoscopic surgery. Other minimally invasive treatment measures can be applied at the same time. For patients who can not tolerate surgical treatment, puncture, endoscopy, intervention and other means can be selected, combined with drugs for conservative treatment, so as to obtain the opportunity of surgical treatment or improve the prognosis. In this paper, the latest research literature at home and abroad has been collected to review and summarize the latest research progress in the physiology and aging characteristics of the biliary tract in the elderly, the diagnosis and treatment of gallstones and acute calculous cholecystitis. It is hoped that this paper can provide reference for the clinical diagnosis and treatment of such diseases, and think and prospect the future research.

Cholecystolithiasis is a common clinical disease, which can be secondary to cholecystitis, cholangitis, pancreatitis and gallbladder cancer. Many patients with gallstone have no obvious clinical symptoms such as biliary colic. Some patients are indeed asymptomatic because there are not obstruction and inflammation. However, some patients have no symptoms due to ignorance, mistaken for stomach pain, enteritis, etc. Some patients have no symptoms because of insensitivity and insufficient perception. For other patients with abnormal gallbladder morphology and structure, non-functional gallbladder must be no pain. To clarify the concept, diagnosis and classification of asymptomatic gallstones according to pathophysiological stages is conducive to the development of personalized management strategies. According to guidelines formulated by different periods, different regions and different medical institutions, there are many controversies regarding the management of asymptomatic gallstones, including regular follow-up, drug treatment, surgical resection or gallblades-saving lithotomy. Reasonable personalized diagnosis and treatment requires more in-depth basic research evidence.

Gallstone is a common and frequent disease and frequent incidence, secondary infection and cancer seriously affect the health of patients. Academic organizations in different regions have issued multiple guidelines and consensus to promote the normative diagnosis and treatment of gallstones. However, in clinical practice, most symptomatic gallstones are treated, while the formation and prevention process of gallstones are ignored, making the concept of treating without a disease has not been strengthened.This article reviews the risk factors and mechanisms of gallstone formation, and points out the importance of effective prevention during stone formation. In the stage of gallstone formation, the high risk factors of stone formation can be analyzed through two aspects of injury factors and protective factors, and the high risk groups of stone formation can be screened out. According to the pathophysiological progression of gallstones, personalized prevention and follow-up strategies can be developed for the stone formation stage of gallstones.

The application of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (Cas) can be limited due to a lack of compatible protospacer adjacent motif (PAM) sequences in the DNA regions of interest. Recently, SpRY, a variant of Streptococcus pyogenes Cas9 (SpCas9), was reported, which nearly completely fulfils the PAM requirement. Meanwhile, PAMs for SpRY have not been well addressed. In our previous study, we developed the PAM Definition by Observable Sequence Excision (PAM-DOSE) and green fluorescent protein (GFP)‍-reporter systems to study PAMs in human cells. Herein, we endeavored to identify the PAMs of SpRY with these two methods. The results indicated that 5'-NRN-3', 5'-NTA-3', and 5'-NCK-3' could be considered as canonical PAMs. 5'-NCA-3' and 5'-NTK-3' may serve as non-priority PAMs. At the same time, PAM of 5'-NYC-3' is not recommended for human cells. These findings provide further insights into the application of SpRY for human genome editing.
CRISPR-Associated Protein 9/metabolism* ; CRISPR-Cas Systems ; DNA ; Gene Editing/methods* ; Humans ; Streptococcus pyogenes/metabolism*

OBJECTIVE: To review the development of adult and pediatric liver transplantation in Tianjin First Center Hospital, and to enhance academic exchanges, improve technological innovation, and jointly promote the progress and maturity in the field of liver transplantation. METHODS: The development of liver transplantation in Tianjin First Center Hospital was analyzed. The clinical data of adult and pediatric liver transplantation from September 1998 to September 2018 were collected. The important events and technological innovation achievements of liver transplantation during the 20 years were summarized. RESULTS: The first clinical liver transplantation was attempted in Tianjin First Central Hospital in April 1980. The first long-term survival adult liver transplantation in China was completed in 1994 (11 years survival after the operation). The specialized team of liver transplantation was formally established in September 1998. The 20-year clinical exploration and progress reflected the characteristics of era changes and technological innovation during the rapid development of liver transplantation in China. Our center performed liver re-transplantation in January 1999, reduced-size pediatric liver transplantation in August 2000. In May 2001, we organized the formulation for the preventive and treatment plan for hepatitis B recurrence after liver transplantation. We performed combined liver and kidney transplantation in July 2002, split liver transplantation (SLT) in April 2004, the first domino liver transplantation (DLT) in August 2005. Pediatric living donor liver transplantation (LDLT) was initiated in October 2006, adult LDLT was carried out in August 2007. In September 2007, the first living donor combined liver and kidney transplantation from the same donor in Asia was performed. The first domino+living donor double grafts liver transplantation in the world was performed in January 2009. In March 2011, we performed laparoscopically assisted right hepatic lobe liver transplantation (LDLT) with middle hepatic vein. In May 2014, living donor laparoscopic left lateral lobe procurement was successfully established. In April 2016, simultaneous liver, pancreas and kidney multi-organ transplantation was completed. Domino donor-auxiliary liver transplantation was performed in February 2017. In December 2017, extracorporeal membrane oxygenation (ECMO)-supported liver transplantation in a patient with severe pulmonary hypertension was successfully completed. Liver transplantation combined with partial splenectomy was established in April 2018. Cross-domino liver transplantation (hypersensitive kidney transplantation with auxiliary liver transplantation+pediatric liver transplantation) was performed in May 2018. During the 20 years, the team has performed or assisted other centers in Beijing, Shanghai, Guangzhou and Shenzhen to carry out more than 10 000 cases of liver transplantations. A total of 7 043 cases of various types of liver transplantation were performed in the single center of the hospital (6 005 adult liver transplantations and 1 038 pediatric liver transplantations). Concerning adult liver transplantation, the cumulative 1-year, 3-year and 5-year survival rate from September 1998 to March 2003 were 83.1%, 73.0% and 69.0%, from April 2003 to March 2009 were 85.3%, 76.2% and 72.1% and from April 2009 to September 2018 were 87.5%, 79.2% and 75.1%, respectively. The cumulative 1-year, 3-year and 5-year survival rate for pediatric liver transplantation were 93.5%, 92.2% and 90.2%, respectively. The nucleoside (acid) analogue combined with low dose hepatitis B immunoglobulin (HBIG) was developed to prevent the recurrence of hepatitis B after liver transplantation, this plan has reduced the recurrence rate of hepatitis B and the 5-year re-infection rate of hepatitis B virus (HBV) after liver transplantation significantly. The risk assessment system for tumor recurrence after liver transplantation was established and individual treatment method was established based on this assessment system. Continuous exploration and improvement of liver transplantation for liver cancer, liver re-transplantation, liver transplantation with portal vein thrombosis, SLT, DLT and multi-organ combined transplantation have significantly improved the clinical efficacy of patients and the post-operative survival rate. CONCLUSIONS The liver transplantation team of Tianjin First Center Hospital has carried out a scientific and technological exploration on the key problems and technical difficulties of clinical liver transplantation. This work strongly has initiated and promoted the rapid development of liver transplantation in China. The restrictive barrier of hepatitis B recurrence after liver transplantation has been overcome. The risk prevention and control system of tumor recurrence after liver transplantation has been established. A series of innovative achievements that can be popularized have been achieved in the field of complex liver transplantation and expansion of donor liver source. The iterative progress and sustainable development of liver transplantation have been realized.
China ; Humans ; Liver Transplantation