Clinical efficacy of combined therapy in children with stage 4 neuroblastoma / 中国当代儿科杂志

OBJECTIVES@#To study the early clinical efficacy of combined therapy of stage 4 neuroblastoma.@*METHODS@#A retrospective analysis was performed on the medical data and follow-up data of 14 children with stage 4 neuroblastoma who were diagnosed in Hong Kong University-Shenzhen Hospital from January 2016 to June 2021.@*RESULTS@#The median age of onset was 3 years and 7.5 months in these 14 children. Among these children, 9 had positive results of bone marrow biopsy, 4 had N-Myc gene amplification, 13 had an increase in neuron-specific enolase, and 7 had an increase in vanilmandelic acid in urine. Based on the results of pathological examination, differentiated type was observed in 6 children, undifferentiated type in one child, mixed type, in one child and poorly differentiated type in 6 children. Of all the children, 10 received chemotherapy with the N7 regimen (including 2 children receiving arsenic trioxide in addition) and 4 received chemotherapy with the Rapid COJEC regimen. Thirteen children underwent surgery, 14 received hematopoietic stem cell transplantation, and 10 received radiotherapy. A total of 8 children received Ch14.18/CHO immunotherapy, among whom 1 child discontinued due to anaphylactic shock during immunotherapy, and the other 7 children completed Ch14.18/CHO treatment without serious adverse events, among whom 1 child was treated with Lu177 Dotatate 3 times after recurrence and is still undergoing chemotherapy at present. The median follow-up time was 45 months for all the 14 children. Four children experienced recurrence within 2 years, and the 2-year overall survival rate was 100%; 4 children experienced recurrence within 3 years, and 7 achieved disease-free survival within 3 years.@*CONCLUSIONS@#Multidisciplinary combined therapy is recommended for children with stage 4 neuroblastoma and can help them achieve better survival and prognosis.

Evaluation of short-term clinical effect of minimally invasive total hip arthroplasty with direct anterior approach in lateral position / 中国骨伤

OBJECTIVE@#To investigate the short-term clinical effect of direct anterior approach (DAA) in total hip arthroplasty(THA).@*METHODS@#From January 2018 to September 2018, the data of 30 patients(33 hips) who underwent the first THA using the side lying DAA completed by the same operation team were followed up and evaluated. There were 19 males and 11 females;the age was 58 to 80 (69.0±5.4) years old;the visual analogue scale (VAS) of pain was used, Harris scoring system, operation time, intraoperative blood loss, related complications and hip radiographs were evaluated in clinical and imaging aspects.@*RESULTS@#Thirty patients (33 hips) were followed up for 12 to 20(14.3±3.7) months, operation time (66.0±7.2) min and intraoperativehemorrhage (156±32) ml. The position of acetabulum prosthesis was examined by imaging:anteversion angle (18.6±3.6)° and abduction angle (41.2±4.8)° respectively. The VAS score was improved from 7 to 9(8.1±1.4) before operation to 1 to 3(1.9±0.7) at 1 month after operation. Harris score of hip joint improved significantly, from 28 to 46(35.4±5.2) before operation to 76 to 92 (88.6±4.5) at 1 month after operation, 74 to 93 (85.6±6.9) at 6 months after operation, and 79 to 95 (90.7±8.1) at 12 months after operation, the difference was statistically significant(<0.05). Complications occurred in 3 cases of fracture of the proximal femur, including 1 case of hip sprain fracture due to careless walking one month after operation. Considering that incomplete fracture may have occurred during the operation, 1 case of avulsion fracture of anterior inferior iliac spine, no deep infection, no dislocation. There were 1 case of injury of lateral femoral cutaneous nerve and 2 cases of injury of tensor fascia lata, among which 1 case was complete incision of the edge of the hook.@*CONCLUSION@#The primary THA with DAA in lateral position has a good short;term clinical effect, can meet the needs of patients' rapid recovery, and is a safe and effective surgical approach.

Clinical study of Jiawei Xiaoyao Powder(JWXYP) on preventing delirium in elderly patients with hip fracture after operation / 中国骨伤

OBJECTIVE@#To study the effect of soothing liver, relieving depression, invigorating spleen and reinforcing blood on reducing delirium in elderly patients with hip fracture.@*METHODS@#From December 2014 to June 2018, 180 elderly patients with hip fracture admitted were divided into treatment group and placebo group according to the order of admission:90 patients in treatment group were treated with Jiawei Xiaoyao Powder(JWXYP), including 32 males and 58 females, with an average age of(72.12±4.92), involving 67 cases of femoral trochanter fractures and 23 cases of femoral neck fractures; 35 cases underwent dynamic hip screw fixation, 31 cases underwent intramedullary fixation and 24 cases underwent artificial hip replacement. In the placebo group, 90 patients were treated with placebo, including 37 males and 53 females, with an average age of(72.91±5.43) years old, involving 69 cases of femoral trochanteric fractures and 21 cases of femoral neck fractures, including 37 cases underwent dynamic hip screw fixation, 30 cases underwent intramedullary fixation and 23 cases underwent artificial hip replacement. The age, sex, injury site, intraoperative bleeding volume, postoperative drainage, operation time, anesthesia time, post-operative pain score, post-operative hemoglobin, post-operative CRP, delirium severity(DRS) score and delirium occurrence were observed and compared between the two groups.@*RESULTS@#All patients were followed up until delirium returned to normal, postoperative delirium was found in 12 cases (13.33%) in the treatment group and in 39 cases(43.33%) in the placebo group, the treatment group was significantly better than the placebo group. The monitoring indexes of the two groups were compared:post-operative pain score(=0.002), post-operative hemoglobin(=0.012), post-operative CRP(=0.042).@*CONCLUSIONS@#JWXYP can relieve liver depression, invigorate spleen and invigorate blood circulation, reduce pain, inflammatory stimulation and supplement blood volume after operation, and significantly reduce the incidence of delirium after operation.

Antioxidant xanthones and anthraquinones isolated from a marine-derived fungus Aspergillus versicolor / 中国天然药物

Chemical examination of an EtOAc extract of cultured Aspergillus versicolor fungus from deep-sea sediments resulted in the isolation of four xanthones, eight anthraquinones and five alkaloids, including a new xanthone, oxisterigmatocystin D (1) and a new alkaloid, aspergillusine A (13). High resolution electron impact mass spectrometry (HR-EI-MS), FT-IR spectroscopy, and NMR techniques were used to elucidate the structures of these compounds, and the absolute configuration of compound 1 was established by its NMR features and coupling constant. Furthermore, the biosynthesis pathway of these xanthones and anthraquinones were deduced, and their antioxidant activity and cytotoxicity in human cancer cell lines (HTC-8, Bel-7420, BGC-823, A549, and A2780) were evaluated. The trolox equivalent antioxidant capacity (TEAC) assay indicated most of the xanthones and anthraquinones possessing moderate antioxidant activities. The Nrf2-dependent luciferase reporter gene assay revealed that compounds 6, 7, 9, and 12 potentially activated the expression of Nrf2-regulated gene. In addition, compounds 5 and 11 showed weak cytotoxicity on A with the IC values of 25.97 and 25.60 μmol·L, respectively.

Treatment of Schatzker IV tibial plateau fractures with arthroscopy combined with MIPPO technique / 中国骨伤

<p><b>OBJECTIVE</b>To discusses the clinical effects of arthroscopy combined with minimally invasive percutaneous plate osteosynthesis(MIPPO) technology in treating Schatzker IV tibial plateau fractures.</p><p><b>METHODS</b>From January 2012 to January 2016, 19 patients with Schatzker type IV tibial plateau fractures were treated with arthroscopy combined with minimally invasive technique including 12 males and 7 females with an average age of 46.5 years old ranging from 19 to 78 years old. Patients were suffering knee pain, swelling, flexion and extension limited, and other symptoms preoperative. Patients were followed up and assessed by Rasmussen knee function score.</p><p><b>RESULTS</b>No infection, traumatic arthritis, and knee joint valgus occurred after operation. Nineteen cases were followed up for 12 to 24 months with an average of 18.6 months. Fracture healing time was 3 to 5 months with an average of 3.8 months. The knee pain and limited mobility improved significantly. The range of autonomic movement of joints was from 90 to 136 degrees. According to Rasmussen functional score criteria, the total score was 27.00±2.49, the result was excellent in 16 cases, good in 2 cases, fair in 1 case.</p><p><b>CONCLUSIONS</b>Arthroscopic treatment for Schatzker type IV tibial plateau fractures combined with MIPPO can simultaneously treat internal structural injuries such as meniscus and other knee joints, with less trauma, fewer complications, and faster joint function recovery, but we must strictly grasp surgical indications and avoid expanding injuries.</p>

Antioxidant xanthones and anthraquinones isolated from a marine-derived fungus Aspergillus versicolor / 中国天然药物

Chemical examination of an EtOAc extract of cultured Aspergillus versicolor fungus from deep-sea sediments resulted in the isolation of four xanthones, eight anthraquinones and five alkaloids, including a new xanthone, oxisterigmatocystin D (1) and a new alkaloid, aspergillusine A (13). High resolution electron impact mass spectrometry (HR-EI-MS), FT-IR spectroscopy, and NMR techniques were used to elucidate the structures of these compounds, and the absolute configuration of compound 1 was established by its NMR features and coupling constant. Furthermore, the biosynthesis pathway of these xanthones and anthraquinones were deduced, and their antioxidant activity and cytotoxicity in human cancer cell lines (HTC-8, Bel-7420, BGC-823, A549, and A2780) were evaluated. The trolox equivalent antioxidant capacity (TEAC) assay indicated most of the xanthones and anthraquinones possessing moderate antioxidant activities. The Nrf2-dependent luciferase reporter gene assay revealed that compounds 6, 7, 9, and 12 potentially activated the expression of Nrf2-regulated gene. In addition, compounds 5 and 11 showed weak cytotoxicity on A with the IC values of 25.97 and 25.60 μmol·L, respectively.

Clinical research of percutaneous K-wires fixation after manipulative reduction combining with gypsum or splint fixation for treatment of the Barton fractures of aged people / 中国骨伤

<p><b>OBJECTIVE</b>To investigate the feasibility of using percutaneous K-wires fixation after manipulative reduction combining with gypsum or splint fixation for treatment of the Barton fracture of aged people.</p><p><b>METHODS</b>From June 2011 to May 2013, 91 patients with closed and fresh Barton fracture were analyzed retrospectively which information was complete. According to different methods these patients were divided into three groups: 33 patients in group A were treated by simplex fixation of gypsum or splint after manipulative reduction, including 20 males and 13 females with an average age of (63.27 ± 5.83) years old; 35 patients in group B were treated by percutaneous K-wires fixation after manipulative reduction combining with gypsum or splint fixation, including 23 males and 12 females with an average age of (64.25 ± 4.53) years old; 23 patients in group C were treated by open reduction and internal fixation including 15 males and 8 females with an average age of (64.04 ± 4.83) years old.</p><p><b>RESULTS</b>All the patients were followed up for 10 to 16 months with an average of 12 months by the method of recheckging in outpatient clinic. According Dienst scoring system: in group A, the result was excellent in 15 cases, good in 7 cases, fair in 11 cases; in group B, excellent in 25 cases, good in 6 cases, fair in 4 cases; in group C, excellent in 16 cases, good in 5 cases, fair in 2 cases. The excellent and good rate of group C was better than that of group B and A, further more group B is better than group A. According to X- ray standard, in group A, 26 patients were acceptable, 7 patients were unacceptable; in group B,28 patients were acceptable, 7 patients were uacceptable; in group C, 19 patients were acceptable,4 patients were unacceptable; there were no statistical significant difference (P > 0.05). There were 18 patients with emerging recrispation in group A, 10 patients in group B, 6 patients in group C, the result of group B and C was better than that of group A, and there was no statistically significant between group B and C (P > 0.05).</p><p><b>CONCLUSION</b>Using percutaneous K-wires fixation after manipulative reduction combining with gypsum or splint fixation for treatment of the Barton fracture of aged people is an effective method, it has advantages of micro-invasived, reliable fixation, less complication, not need to fix wrist joint and early functional exercise.</p>

Investigation of operative process in treating lumbar intervertebral disc protrusion in aged patients / 中国骨伤

<p><b>OBJECTIVE</b>To investigate the operative management and surgical techniques for lumber disc herniation (LDH) in aged patients (>or=65 years).</p><p><b>METHODS</b>From 2005 to 2010,the data of 43 patients with LDH underwent surgical treatment were retrospectively analyzed. There were 25 males and 18 females,aged from 65 to 70 years old with an average of 67.6 years. The course of disease was from 6 weeks to 7 years with an average of 10.2 years. Fenestratiodn discectomy or extended fenestration discectomy and unilateral or bilateral fenestration were used according to the conditions of location,type of herniated macleos polposus and nerve root compression. Among the patients,the nerve root canal was enlarged,hyperplastic osteophyma and soft tissue were removed, bilateral articular process was reserved in order to maintain the stability of the lumbar segment.</p><p><b>RESULTS</b>All the patients were followed up from 1.2 to 5.2 years. According to the criteria of HU you-gu, 25 cases got excellent results, 15 good, 2 fair and 1 poor. No infection or nerve injury was found. Nerve function of patients had a normal or near normal recovery.</p><p><b>CONCLUSION</b>For the treatment of LDH in aged patents,it is key that reasonably choose the operative method, completely remove the hyperplasy of diseased region and enlarge the nerve root canal, thoroughly loose the nerve root.</p>

Diagnosis of medial collateral ligament injury by stress X-ray and MRI of knee joint / 中国骨伤

<p><b>OBJECTIVE</b>To study the application of stress X-ray of knee joint for the diagnosis ot medial collateral ligament injury.</p><p><b>METHODS</b>From January 2008 to June 2011, 46 patients with medial collateral ligament injury were reviewed. Among the patients, 32 patients were male and 14 patients were female, ranging in age from 28 to 72 years, with an average of (49.46 +/- 22.54) years. Left knee joint injuries occurred in 22 patients, and 24 patients had injuries in right knee joint. Thirty-one patients were treated with surgical method, and 15 patients were treated with conservative method including 1 patient finally received operation after 3 months treatment with conservative method without effect. The injury side and normal side were observed by X-ray under stress and non-stress, as well as MRI. Operation was used to treat patients with medial gap markedly widen and the deep layer of medial collateral ligament injury in MRI. A line (line A) was drawn from medial condyle to lateral condyle of tibia platform. The line A was translated to the inner margin of condylus medialis femoris, which was line B. The perpendicular distance was measured, which was line C. The ration of space difference (R) was calculated by (space at stress force of injured side-space at non-stress force of injured side)/(space at stress force of normal side-space at non-stress force of normal side). The relationship between R and ligament injury type were investigated.</p><p><b>RESULTS</b>There were 17 patients with injuries of superficial lamella, 21 patients with injuries of deep lamella. Eight patients had associated injuries of articular capsule and posterior cruciate ligament. When R value was between 1.51 to 5.24, the injury of superficial ligament was found in 15 patients, actual injuries were found in 17 patients. When R value between 5.28 and 13.85, the injuries of bathypelagic ligament were found in 19 patients, actual injuries were found in 21 patients. When R value was between 15.61 and 26.25, the associated injuries of articular capsule and posterior cruciate ligament were found in 7 patients, actual injuries were found in 8 patients.</p><p><b>CONCLUSION</b>Using the specific value as the standard of measurement can provide the quantizatily fractionation of the medial collateral ligament injury.</p>

Diagnosis and treatment of tarsometatarsal joint complex injury / 中国骨伤

<p><b>OBJECTIVE</b>To explore the diagnosis and treatment of tarsometatarsal joint complex injury (TJC).</p><p><b>METHODS</b>From January 2007 to January 2009,16 patients with tarsometatarsal joint complex injury were treated with open reduction and internal fixation. There were 12 males and 4 females, ranging in age from 21 to 45 years with an average of 34.1 years. Seven cases were left and 9 cases were right and all injuries caused by direct violence. Four cases caused by traffic accident 5 by fall from high and 7 by crush injury. Intercuneiform dislocation were in 11 cases, naviculocuneiform joint dislocation in 3 cases and cuboid fracture in 2 cases. All the cases were three column injuries. According to the situation of exploring and the stability, screw fixation was used for intertarsal joint, internal and middle column tarsometatarsal joint, the Kirschner wire fixation for external column and miniature plate fixation for comminuted fracture of metatarsal bones and compressible fracture of cuboid. The criteria of the AOFAS Foot and Ankle Surgery by the United States Association of ankle-rear foot functional scale was used to evaluate the clinical effect.</p><p><b>RESULTS</b>All the patients were followed up,the duration ranged from 6 to 18 months(averaged 12.6 months). According to the score system of AOFAS,the total score was (74.6+/-10.4 ) points, including pain items of (29.3+/-5.9), the score of functional items of (32.4+/-5.6) points, and power lines of (12.9+/-2.6). All the incisions were primarily healed without infection, skin necrosis,fixture broken or loosen. Three cases received arthrodesis due to osteoarthritis. Four cases were followed up continually because they only had the radiologic osteoarthritis without pain.</p><p><b>CONCLUSION</b>Anatomical reduction and stable fixation is the key point of the treatment of tarsometatarsal joint complex injury. Open reduction and internal fixation at the first stage is good for secondary arthrodesis.</p>

Treatment of double injuries of superior shoulder suspensory complex / 中国骨伤

<p><b>OBJECTIVE</b>To explore operative method for the treatment of double injuries of superior shoulder suspensory complex (SSSC).</p><p><b>METHODS</b>From January 2008 to March 2010,11 patients wiht SSSC injuries were treated, including 9 males and 2 females with an average age of 38 years (ranged from 20 to 47 years). The patients were treated with 4 methods as follows: 4 patients with fractures of neck of scapula combined with homolateral fracture of clavicle were treated with reconstituted plates; 2 patients with fractures of coracoid process or disruption of coracoclavicular ligament combined with the fracture of extremitas acromialis claviculae or acromioclavicular dislocation were treated with clavicular hook plates and cannulated compression screws; 3 patients with injuries of basilar part of acromial process combined with the glenoid cavity and acromioclavicular articulation were treated with reconstuction plates and clavicular hook plates; 2 patients with fractures of acromial process combined with acromioclavicular dislocation and the fracture of lateral third of clavicle were treated with small "T" plates and clavicular hook plates.</p><p><b>RESULTS</b>Among 11 patients, 9 patients were followed up with an average duration of 9.2 months (ranged from 6 to 12 months). All the fractures were healed without bone nonunion or failure of internal fixators. The average union time was 2.6 months. The profile of articulatio capitis humeri was normal and hibateral articulatio capitis humeri was symmetrical without crispation, descensus, adduct and adtorsion of articulatio capitis humeri or other abnormity. According to the Constant-Murley evaluation system, the score ranged from 69 to 100, with an average of 89.7, which included average pain score of 10 to 15, daily activities score of 14 to 20, myodynamia score of 15 to 25, territory score of 34 to 40.</p><p><b>CONCLUSION</b>The double injuries of SSSC should be treated by types to recover the integrity and constancy of SSSC.</p>

Treatment of tarsometatarsal joint injury combined with metatarsal fracture by open reduction and internal fixation / 中国骨伤

<p><b>OBJECTIVE</b>To evaluate the therapy and the key points of the tarsometatarsal joint injury combined with metatarsal fracture.</p><p><b>METHODS</b>From Jan. 2006 to Jul. 2008,19 patients with tarsometatarsal joint injury combined with metatarsal fracture were treated with opened reduction and internal fixation of Kirschner wire or screws, included 13 males and 6 females with an average age of 38.1 years ranging from 21 to 56 years. The classification of tarsometatarsal joint injury showed that there were 2 cases of inner column injury, 5 cases of inner and medial column injury, 3 cases of lateral and medial column injury, 9 cases of tri-column injury. There were 8 cases of shaft fracture, 7 of neck fracture, 19 of foundation fracture.</p><p><b>RESULTS</b>All the incisions were first stage healed without skin necrosis. The healing time of fracture was 11.2 weeks on average. All the patients were followed-up for 6 to 17 months with an average of 12.8 months. According to the standard of AOFAS, the average score was (84.500 +/- 8.553), the results were excellent in 4 cases, good in 9 cases, fair in 3, and poor in 3. The regular daily life was recovered after 6.4 months, 3 patients suffer from mild osteoarthritis.</p><p><b>CONCLUSION</b>No matter which fixed mode was used, the anatomical reduction was the most important to rebuild arches of the foot and recover medial longitudinal and lateral arch. Rebuilding arches of the foot guaranteed the integrity of the stress point scaffold and avoided the pain and limp. The anatomical reduction of tarsometatarsal joint and metatarsal was also important to rebuild the function of foot.</p>

Error diagnosis and inappropriate treatment for Essex-Lopresti injury in 2 cases / 中国骨伤

<p><b>OBJECTIVE</b>To analyze the reason of error diagnosis and inappropriate treatment of radius head fractures complicating distal radioulnar joint dislocation (Essex-Lopresti injury) and to seek for the reasonable treatment.</p><p><b>METHODS</b>From 2008 to 2009, 2 male patients of radius head fractures complicating distal radioulnar joint dislocation were treated, the age was 56 and 66 years old respectively. The symptom included elbow swelling and limitation of forearm rotation. X-ray showed comminuted fractures of capitulum radius. There were 1 case of Mason type III and 1 cases of type IV. Two patients were treated by resection of capitulum radius and plaster fixation after operation.</p><p><b>RESULTS</b>Both of the 2 cases had limited rotation of forearm and the distal radioulnar joint pain. The X-ray showed that the distal radioulnar joint was separated obviously and the proximal radius was translocated.</p><p><b>CONCLUSION</b>The early diagnosis is very important for the Essex-Lopresti injury. Rebuilding the longth of the radius and reduction fixating the distal radioulnar joint after fixated is an effective treatment while the interosseous membranes can't be repaired and rebuilded.</p>

In vitro suppressive effect of angelica polysaccharide on human cytomegalovirus-induced apoptosis via direct infection in CHRF-288-11 cells / 中国实验血液学杂志

The objective of study was to investigate the in vitro suppressive effect of angelica polysaccharide (APS) on human cytomegalovirus-induced apoptosis via direct infection in CHRF-288-11 cells. HCMV AD169 directly infected CHRF-288-11 were cultured in vitro, APS in different doses were added on day 3 after the infection of virus. Cells of every group were collected at different time points. HCMV DNA of cells were detected by using polymerase chain reaction and the apoptotic cells were examined by using Hoechst staining, MTT assay, DNA fragmentation assay and flow cytometry. The results showed that the APS to some extent inhibited the apoptosis of CHRF cells infected by HCMV in vitro in a dose-dependent manner. The expression of HCMV IEA in CHRF-288-11 cells was found by PCR amplification. Morphology observation, flow cytometry assay and DNA fragmentation assay revealed the existence of apoptosis. With the dose decrease of APS added to the infected CHRF cells, the percentage of apoptotic cells increased. It is concluded that the HCMV AD169 can infect CHRF cells directly in vitro and decrease cell viability. HCMV AD169 infection increases the apoptosis of CHRF cells in time-dependent manner. When APS was added to the CHRF cells infected by HCMV AD169 in vitro, the viability of CHRF cells increase, which indicated that APS to some extent protects the CHRF cells infected by HCMV. APS suppresses the cytomegalovirus-induced apoptosis in CHRF cells directly infected in vitro in dose-dependent manner.

Application of flap or musculocutaneous flap primary transplantation in Pilon fractures of type III / 中国骨伤

<p><b>OBJECTIVE</b>To discuss the value of one stage flap or musculocutaneous flap to repair the soft tissue defect in Pilon fracture.</p><p><b>METHODS</b>Twelve cases with Pilon fracture included 9 male and 3 female with an average age of 39.2 years ranging from 21 to 61 years. All fractures were type III according to Rüedi and Allgöwer. According to Gustilo sysytem, III a were in 4 cases, III b were in 8. A thorough debridement was made before internal fixation. After the internal fixation implanted, the tourniquet was released and a thorough debridement was made again. Be sure of the wound cleansed of all dead and foreign material, the wound was covered with local flap, musculocutaneous or gastrocnemius flaps depending on the size and localization of wound.</p><p><b>RESULTS</b>All the patients were followed up from 6 to 36 months, 18 months in average. The function of ankle was assessed according Mazur system. The results were excellent in 6 cases, good in 3, fair in 2 and poor in 1. No infection or necrosis happened on flaps. Although necrosis happened in the wound margin of two patients, they all healled up by conservative methods.</p><p><b>CONCLUSION</b>Primary closure of soft tissue defect in Pilon fracture using flap or musculocutaneous flap have ability to shorten the treating time and recover the function of ankle. It is important to have a thorough debridement of the dead tissue and free bone.</p>

Human cytomegalovirus induces apoptosis of human promyelocytic leukemic cells via direct infection in vitro / 中国实验血液学杂志

The study was purposed to investigate the proliferation and the suppression effect of human cytomegalovirus (hCMV) on human promyelocyte cell line HL-60, and to study whether hCMV can induce apoptosis of HL-60 via direct infection in vitro and its mechanism. Promyelocyte cell line HL-60 and hCMV AD169 strain were co-cultured. PCR was used to detect the direct infection of HL-60 cells by hCMV IEA expression. The apoptosis cells were analyzed by morphologic observation, DNA ladder formation, flow cytometry with Annexin V/PI stain. The results indicated that hCMV AD169 suppressed the differentiation and proliferation of HL-60 cells in vitro significantly (P < 0.05). The suppression was dose-dependent. hCMV DNA was successfully detected in HL-60 cells of viral infection groups by PCR. The apoptotic cells were confirmed by morphologic observation and DNA ladder formation. The results of flow cytometry showed that the percentage of apoptotic cells increased along with the increase of hCMV titer and the time after infection. It is concluded that the promyelocyte can be infected directly by hCMV AD169 strain. hCMV AD169 strain inhibited the differentiation and proliferation of promyelocyte. The apoptosis of HL-60 cells can be induced by hCMV infection. With the increase of viral infectious titer and the time after infection, the percentage of apoptotic cells also increase and produce in dose-dependent and time- dependent manner. Induced apoptosis may be one of the mechanisms of granulocytopenia induced by hCMV infection.

Effect of human cytomegalovirus on hematopoietic system / 中华儿科杂志

<p><b>OBJECTIVE</b>To investigate the mechanism and the suppression effect of human cytomegalovirus (HCMV) on hematopoietic system.</p><p><b>METHODS</b>Semi-solid culture system was used to observe the effect of HCMV AD169 strain on colony forming unit granulocyte/macrophage (CFU-GM), CFU-erythroid (CFU-E), CFU-multipotent (CFU-Mix) and CFU-megakaryocyte (CFU-MK) growth. The techniques of in situ polymerase chain reaction (IS-PCR) and polymerase chain reaction (PCR) were used to demonstrate the existence of HCMV DNA in the colony cells of cultured CFU-GM, CFU-Mix, CFU-MK and CFU-E, respectively. The immediate early antigen (IEA) mRNA in CFU-MK and late antigen (LA) mRNA in CFU-E were detected by reverse transcriptase-polymerase chain reaction (RT-PCR). HCMV early protein P52 was detected with immunohistochemical technique.</p><p><b>RESULTS</b>HCMV AD169 suppressed the differentiation and proliferation of CFU-GM, CFU-E, CFU-Mix and CFU-MK in vitro significantly (P < 0.05). The suppression was dose-dependent. HCMV DNA was successfully detected in CFU-GM, CFU-Mix, CFU-MK colony cells from viral infection groups by IS-PCR, and was detected in CFU-E by PCR, while it was negative in blank control or mock control groups. CFU-MK colony cells expressed HCMV IEA mRNA with the size of 340 bp in virus infection groups of 10(3) plague forming unit (PFU), 10(4) PFU and 10(5) PFU, respectively. The HCMV LA mRNA was detected by RT-PCR and was 263 bp long in positive control group of HCMV-infected human embryonic fibroblasts. The expression of HCMV LA mRNA in CFU-E was negative. The early protein P52 of HCMV in 10(4) PFU group was also identified by immunohistochemical staining.</p><p><b>CONCLUSION</b>HCMV AD169 strains inhibited the differentiation and proliferation of CFU-GM, CFU-E, CFU-Mix and CFU-MK by the infection of the hematopoietic progenitors. HCMV might cause the suppression of hematopoiesis by direct infection, which is thought to be one of the reasons of HCMV infection associated with thrombocytopenia, neutropenia and anemia.</p>